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AU2004218808A1 - Pharmaceutical preparations comprising acid-stabilised insulin - Google Patents

Pharmaceutical preparations comprising acid-stabilised insulin Download PDF

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Publication number
AU2004218808A1
AU2004218808A1 AU2004218808A AU2004218808A AU2004218808A1 AU 2004218808 A1 AU2004218808 A1 AU 2004218808A1 AU 2004218808 A AU2004218808 A AU 2004218808A AU 2004218808 A AU2004218808 A AU 2004218808A AU 2004218808 A1 AU2004218808 A1 AU 2004218808A1
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Australia
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alkyl
aryl
pharmaceutical composition
composition according
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AU2004218808A
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Palle Jakobsen
Niels C. Kaarsholm
Svend Ludvigsen
Peter Madsen
Helle Birk Olsen
Soren Ostergaard
Anders Klarskov Petersen
Gerd Schluckebier
Dorte Bjerre Steensgaard
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Novo Nordisk AS
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Novo Nordisk AS
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    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
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Description

WO 2004/080480 PCT/DK2004/000158 PHARMACEUTICAL PREPARATIONS COMPRISING ACID-STABILISED INSULIN. FIELD OF THE INVENTION The present invention discloses pharmaceutical preparations comprising ligands for the 5 HisB10 Zn 2 1 sites of the R-state insulin hexamer and acid-stabilsed insulin analogues. The compositions release insulin slowly following subcutaneous injection. BACKGROUND OF THE INVENTION Insulin Allostery. The insulin hexamer is an allosteric protein that exhibits both positive and negative cooperativity and half-of-the-sites reactivity in ligand binding. This allosteric behav 10 iour consists of two interrelated allosteric transitions designated LAo and- L 0 , three inter converting allosteric conformation states (eq. 1), LAO LBo
T
6
T
3
R
3 R6 (1) 15 designated T 6 , T 3
R
3 , and R 6 and two classes of allosteric ligand binding sites designated as the phenolic pockets and the Hiss"o anion sites. These allosteric sites are associated only with insulin subunits in the R conformation. Insulin Hexamer Structures and Ligand Binding. The T- to R-transition of the insulin 20 hexamer involves transformation of the first nine residues of the B chain from an extended conformation in the T-state to an a-helical conformation in the R-state. This coil-to-helix transition causes the N-terminal residue, Phe B1 , to undergo an - 30 A change in position. This conformational change creates hydrophobic pockets (the phenolic pockets) at the sub unit interfaces (three in T 3
R
3 , and six in R), and the new B-chain helices form 3-helix bun 25 dies (one in T 3
R
3 and two in R) with the bundle axis aligned along the hexamer three-fold symmetry axis. The His
B
' Zn 2 in each R 3 unit is forced to change coordination geometry from octahedral to either tetrahedral (monodentate ligands) or pentahedral (bidentate ligands). Formation of the helix bundle creates a narrow hydrophobic tunnel in each R 3 unit that extends from the surface -12 A down to the HisBo metal ion. This tunnel and the His
B
"" 30 Zn 24 ion form the anion binding site. Hexamer Ligand Binding and Stability of Insulin Formulations. The in vivo role of the T to R transition is unknown. However, the addition of allosteric ligands (e.g. phenol and chloride ion) to insulin preparations is widely used. Hexamerization is driven by coordination of Zn 2
+
WO 2004/080480 PCT/DK2004/000158 2 at the His B 10 sites to give Ts, and the subsequent ligand-mediated transition of T 6 to TaRa and to R 6 is known to greatly enhance the physical and chemical stability of the resulting formula tions. Ligand Binding and Long Acting Insulin Formulations. Although the conversion of T 6 to T 3
R
3 5 and R 6 improves the stability of the preparation, the rate of absorption following subcutane ous injection of a soluble hexameric preparation is not much affected by the addition of phe nol and cloride. Putative events following injection of a soluble hexameric preparation. The small molecule ligands initially diffuse away from the protein. The affinity of the ligands for insulin may help 10 to slow this process. On the other hand, the affinity of Zn 2 + for e.g. albumin and the large ef fective space available for diffusion of the lipophilic phenol will tend to speed up the separa tion. In about 10-15 minutes after injection, the distribution of insulin species in the subcuta neous tissue will roughly correspond to that of a zinc-free insulin preparation at the same di lution. Then, the equilibrium distribution of species at this point will determine the observed 15 absorption rate. In this regimen, absorption rates vary between about 1 hour (for rapid-acting insulin analogues, such as Asp 8 28 human insulin) and about 4 hours (Co 3 -hexamer). Current Approaches Toward Slow Acting Insulins. The inherent limitation of the absorption half-life to about 4 hours for a soluble human insulin hexamer necessitates further modifica tions to obtain the desired protraction. Traditionally, this has been achieved by the use of 20 preparations wherein the constituent insulin is in the form of a crystalline and/or amorphous precipitate. In this type of formulation, the dissolution of the precipitate in the subcutaneous depot becomes rate-limiting for the absorption. NPH and Ultralente belong to this category of insulin preparations where crystallization/precipitation is effected by the addition of protamine and excessive zinc ion, respectively. 25 Another approach involves the use of insulin derivatives where the net charge is increased to shift the isoelectric point, and hence the pH of minimum solubility, from about 5.5 to the physiological range. Such preparations may be injected as clear solutions at slightly acidic pH. The subsequent adjustment of the pH to neutral induces crystallization/precipitation in the subcutaneous depot and dissolution again becomes rate-limiting for the absorption. 30 Glym 1 ArgB 31 ArgB3 2 human insulin belongs to this category of insulin analogues. Most recently, a series of soluble insulin derivatives with a hydrophobic moiety covalently at tached to the side chain of Lys
B
" have been synthesized. These derivatives may show pro longed action profile due to various mechanisms including albumin binding (e.g. B29-N ' myristoyl-des(B30) human insulin), extensive protein self-association and/or stickiness (e.g.
WO 2004/080480 PCT/DK2004/000158 3 B29-N'-(N-lithocholyl-y-glutamyl)-des(B30) human insulin) induced by the attached hydro phobic group. SUMMARY OF THE INVENTION The present invention provides a pharmaceutical preparation comprising ligands for 5 the His B 0 Zn2+ sites of the R-state insulin hexamer, zinc ions and acid-stabilised insulin ana logs. The preparations form clear solutions at slightly acidic pH. When the pH is adjusted to wards neutral upon subcutaneous injection, the ligands work to stabilize hexamers and mod ify solubility in the neutral pH range. As a result, the preparations release insulin slowly fol lowing subcutaneous injection. 10 The invention furthermore provides a method of preparing ligands for the Hiss"o Zn 2 " sites of the R-state insulin hexamer comprising the steps of * Identifying a starter compound that binds to the R-state Hissio-Zn2+ site *optionally attaching a fragment consisting of 0 to 5 neutral a- or 1-amino acids 15 *attaching a fragment comprising 1 to 20 positively charged groups independently se lected from amino or guanidino groups. The invention also provides a method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to the acid stabilised insulin preparation. 20 The invention finally provides a method of treating type 1 or type 2 diabetes comprising ad ministering toa patient in need thereof a theraputically effective amount of a pharmaceutical preparation of the invention. DEFINITIONS 25 The following is a detailed definition of the terms used to describe the invention: "Halogen" designates an atom selected from the group consisting of F, CI, Br and I. The term "Cl-C 6 -alkyl" as used herein represents a saturated, branched or straight hydrocar bon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 30 isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl and the like. The term "Cl-C6-alkylene" as used herein represents a saturated, branched or straight bivalent hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but WO 2004/080480 PCT/DK2004/000158 4 are not limited to, methylene, 1,2-ethylene, 1,3-propylene, 1,2-propylene, 1,4-butylene, 1,5 pentylene, 1,6-hexylene, and the like. The term "C 2
-C
6 -alkenyl" as used herein represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one double bond. Examples of such 5 groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3-buta dienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 1-pentenyl, 2-pentenyl, 3 pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5 hexenyl and the like. The term "C 2
-C
6 -alkynyl" as used herein represents a branched or straight hydrocarbon 10 group having from 2 to 6 carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4 hexynyl, 5-hexynyl, 2,4-hexadlynyl and the like. The term "C 1
-C
6 -alkoxy" as used herein refers to the radical -O-Cl-C 6 -alkyl, wherein Cl-C 6 -alkyl 15 is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, sec-butoxy, tert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like. The term "C 3
-C
8 -cycloalkyl" as used herein represents a saturated, carbocyclic group having from 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like. 20 The term "C"-cycloalkenyl" as used herein represents a non-aromatic, carbocyclic group hav ing from 4 to 8 carbon atoms containing one or two double bonds. Representative examples are 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3 cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1,4-cyclooctadienyl and the like. 25 The term "heterocyclyl" as used herein represents a non-aromatic 3 to 10 membered ring con taining one or more heteroatoms selected from nitrogen, oxygen and sulphur and optionally containing one or two double bonds. Representative examples are pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like. The term "aryl" as used herein is intended to include carbocyclic, aromatic ring systems such 30 as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, carbocyclic, aromatic ring systems. Representative examples are phenyl, biphenylyl, naphthyl, anthracenyl, phe nanthrenyl, fluorenyl, indenyl, azulenyl and the like. Aryl is also intended to include the par tially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4 35 dihydronaphthyl and the like.
WO 2004/080480 PCT/DK2004/000158 5 The term "arylene" as used herein is intended to include divalent, carbocyclic, aromatic ring systems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, divalent, carbocyclic, aromatic ring systems. Representative examples are phenylene, biphenylylene, naphthylene, anthracenylene, phenanthrenylene, fluorenylene, indenylene, azulenylene and 5 the like. Arylene is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated deriva tives are 1,2,3,4-tetrahydronaphthylene, 1,4-dihydronaphthylene and the like. The term "aryloxy" as used herein denotes a group -O-aryl, wherein aryl is as defined above. The term "aroyl"' as used herein denotes a group -C(O)-aryl, wherein aryl is as defined above. 10 The term "heteroaryl" as used herein is intended to include aromatic, heterocyclic ring sys tems containing one or more heteroatoms selected from nitrogen, oxygen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyc lic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sul phur. Representative examples are furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, 15 thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3 oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4 thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, ben zofuryl, benzothienyl, indazolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, 20 benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl, thiazolidinyl, 2 thiooxothiazolidinyl and the like. Heteroaryl is also intended to include the partially hydrogen ated derivatives of the ring systems enumerated above. Non-limiting examples of such par tially hydrogenated derivatives are 2,3-dihydrobenzofuranyl, pyrrolinyl, pyrazolinyl, indolinyl, 25 oxazolidinyl, oxazolinyl, oxazepinyl and the like. The term "heteroarylene" as used herein is intended to include divalent, aromatic, heterocyc lic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sul phur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy 30 gen and sulphur. Representative examples are furylene, thienylene, pyrrolylene, oxa zolylene, thiazolylene, imidazolylene, isoxazolylene, isothiazolylene, 1,2,3-triazolylene, 1,2,4 triazolylene, pyranylene, pyridylene, pyridazinylene, pyrimidinylene, pyrazinylene, 1,2,3 triazinylene, 1,2,4-triazinylene, 1,3,5- triazinylene, 1,2,3-oxadiazolylene, 1,2,4-oxadiazolylene, 1,2,5-oxadiazolylene, 1,3,4-oxadiazolylene, 1,2,3-thiadiazolylene, 1,2,4-thiadiazolylene, 1,2,5 35 thiadiazolylene, 1,3,4-thiadiazolylene, tetrazolylene, thiadiazinylene, indolylene, isoindolylene, WO 2004/080480 PCT/DK2004/000158 6 benzofurylene, benzothienylene, indazolylene, benzimidazolylene, benzthiazolylene, ben zisothiazolylene, benzoxazolylene, benzisoxazolylene, purinylene, quinazolinylene, quinoliz inylene, quinolinylene, isoquinolinylene, quinoxalinylene, naphthyridinylene, pteridinylene, carbazolylene, azepinylene, diazepinylene, acridinylene and the like. Heteroaryl is also in 5 tended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydro benzofuranylene, pyrrolinylene, pyrazolinylene, indolinylene, oxazolidinylene, oxazolinylene, oxazepinylene and the like. The term "ArGI" as used herein is intended to include an aryl or arylene radical as applicable, 10 where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, anthra cenyl, phenanthrenyl, fluorenyl, indenyl, and azulenyl as well as the corrresponding divalent radicals. The term "ArG2" as used herein is intended to include an aryl or arylene radical as applicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, fluo 15 renyl, and indenyl, as well as the corrresponding divalent radicals. The term "Hetl" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3 20 triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, indazolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, di 25 azepinyl, acridinyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding divalent radicals. The term "Het2" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 30 1,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3 triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthia zolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, WO 2004/080480 PCT/DK2004/000158 7 carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding divalent radi cals. The term "Het3" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, 5 pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, pyridyl, tetrazolyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolyl, isoqui nolyl, quinoxalinyl, carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrrespond ing divalent radicals. 10 "Aryl-Cl-C 6 -alkyl", "heteroaryl-Cl-C 6 -alkyl", "aryl-C 2
-C
6 -alkenyl" etc. is intended to mean Cl-C 6 alkyl or C 2
-C
6 -alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example: The term "optionally substituted" as used herein means that the groups in question are either 15 unsubstituted or substituted with one or more of the substituents specified. When the groups in question are substituted with more than one substituent the substituents may be the same or different. Certain of the above defined terms may occur more than once in the structural formulae, and upon such occurrence each term shall be defined independently of the other. 20 Furthermore, when using the terms "independently are" and "independently selected from" it should be understood that the groups in question may be the same or different. The terms "treatment" and "treating" as used herein means the management and care of a patient for the purpose of combating a disease, disorder or condition. The term is intended to include the delaying of the progression of the disease, disorder or condition, the alleviation or 25 relief of symptoms and complications, and/or the cure or elimination of the disease, disorder or condition. The patient to be treated is preferably a mammal, in particular a human being. The term "fragment" as used herein is intended to mean a bivalent chemical group The term "Neutral amino acid" as used herein is intended to mean any natural (codable) and non-natural amino acid, including a- or 1-aminocarboxylic acids, including D-isomers of these 30 (when applicable) without charges at physiologically relevant pH in the side chain, such as glycine, alanine, -alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, aspargine, glutamine, cysteine, methionine, 3-aminobenzoic acid, 4-aminobenzoic acid or the like.
WO 2004/080480 PCT/DK2004/000158 8 The term "positively charged group" as used herein is intended to mean any pharmaceuti cally acceptable group that contains a positive charge at physiologically relevant pH, such as amino (primary, secondary and tertiary), ammonium and guanidino groups. The term "a amino acid" as used herein is intended to mean mean any natural (codable) and 5 non-natural a-aminocarboxylic acid, including D-isomers of these. The term "fl amino acid" as used herein is intended to mean any 1-aminocarboxylic acid, such as -alanine, isoserine or the like. \ The term "desB30" as used herein is intended to mean meant a natural insulin B chain or an analogue thereof lacking the B30 amino acid residue. 10 The amino acid residues are indicated in the three letter amino acid code or the one letter amino code. The terms "BI", "Al" and the like as used herein is intended to mean the amino acid residue in position 1 in the B chain of insulin or analogue thereof (counted from the N-terminal end) and the amino acid residue in position 1 in the A chain of insulin or analogue thereof 15 (counted from the N-terminal end), respectively. When in the specification or claims mention is made of groups of compounds such as car boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, irni dazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia 20 zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, naphthoic acids and salicylic acids, these groups of compounds are in tended to include also derivatives of the compounds from which the groups take their name. The term acid-stabilised insulin as used herein refers to an insulin analog that does not 25 deamidate or dimerize at pH values below 7. Specifically, the analog cannot have Asn or Asp as a C-terminal residue. The term human insulin as used herein refers to naturally produced insulin or recombinantly produced insulin. Recombinant human insulin may be produced in any suitable host cell, for 30 example the host cells may be bacterial, fungal (including yeast), insect, animal or plant cells. The expression "insulin derivative" as used herein (and related expressions) refers to human insulin or an analogue thereof in which at least one organic substituent is bound to one or more of the amino acids. By "analogue of human insulin" as used herein (and related expressions) is meant human 35 insulin in which one or more amino acids have been deleted and/or replaced by other amino WO 2004/080480 PCT/DK2004/000158 9 acids, including non-codeable amino acids, or human insulin comprising additional amino acids, i.e. more than 51 amino acids, such that the resulting analogue possesses insulin ac tivity. The term "phenolic compound" or similar expressions as used herein refers to a chemical 5 compound in which a hydroxyl group is bound directly to a benzene or substituted benzene ring. Examples of such compounds include, but are not limited to, phenol, o-cresol, m-cresol and p-cresol. The term "physiologically relevant pH" as used herein is intended to mean a pH of about 7.1 to 7.9. 10 Abbreviations: 4H3N 4-hydroxy-3-nitrobenzoic acid AcOH acetic acid 15 BT Benzotriazol-5-oyl DMF N,N-Dimethylformamide DMSO Dimethylsulfoxide DIC Diisopropylcarbodiimide EDAC 1-ethyl-3-(3'-dimethylamino-propyl)carbodiimide, hydrochloride 20 Fmoc 9H-Fluorene-9-ylmethoxycarbonyl G, Gly Glycine HOAt 1 -hydroxy-7-azabenzotriazole HOBT 1 -Hydroxybenzotriazole L, Lys Lysine 25 NMP N-methyl-2-pyrrolidone Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl Pmc 2,2,5,7,8-pentamethylchroman-6-sulfonyl R, Arg Arginine TFA Trifluoroacetic acid 30 Abbreviations for non-natural amino acid residues: WO 2004/080480 PCT/DK2004/000158 10 4-Abz O 4-Apac O BT ON 0 N N H H BRIEF DESCRIPTION OF DRAWINGS 5 Fig. 1: The effect of various concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoyl-Arg 1 2
-NH
2 on the pH-dependence of GlyA 1 , AspB 28 insulin solubility. Fig. 2: The effect of a high concentrations of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoyl-Arg ,2
-NH
2 on the pH-dependence of Gly
A
' insulin solubility. Fig. 3: Disappearance from the subcutaneous depot (pig model) of insulin preparations. 10 Curves a)-c) are Gly", AspB 2 6 human insulin formulated with an excess concentration com pared to Zn 2 + of a) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl Arg 3
-NH
2 , b) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyry-Args-NH 2 and c) 4-(2H-Tetrazol-5-yl)benzoyl-Abz-Gly 2 -Args-NH 2 . Curve d) is B29-N'-myristoyl des(B30) human insulin. Curves e) and f) are Gly A 1 human insulin formulated with two dif 15 ferent excess concentrations compared to Zn 2 + of 4-[3-(2H-Tetrazol-5-yl)carbazol-9 ylmethyl]benzoyl-Argl 2
-NH
2 . Fig. 4: 4H3N-assay. UV/vis spectra resulting from a titration of hexameric insulin with the compound 3-hydroxy-2-naphthoic acid in the presence of 4-hydroxy-3-nitrobenzoic acid (4H3N). Inserted in the upper right corner is the absorbance at 444nm vs. the concentration 20 ofligand Fig. 5: TZD-assay. Fluorescence spectra resulting from a titration of hexameric insulin with 5-(3-methoxybenzylidene)thiazolidine-2,4-dione in the presence of 5-(4-dimethylamino benzylidene)thiazolidine-2,4-dione (TZD). Inserted in the upper right corner is the fluores cence at 460 nm vs. the concentration of ligand 25 DESCRIPTION OF THE INVENTION The present invention is based on the discovery that the HisBo Zn" + ligand binding sites of the R-state insulin hexamer can be used to obtain an insulin preparation having prolonged action designed for flexible injection regimes including once-daily, based on acid-stabillised insulins.
WO 2004/080480 PCT/DK2004/000158 11 The basic concept underlying the present invention involves reversible attachment of a ligand to the HisBIo Zn 2 site of the R-state hexamer. A suitable ligand binds to the hexamer metal site with one end while other moieties are covalently attachment to the other end. On this ba sis, prolonged action via modification of preparation solubility may be obtained in a number 5 of ways. However, all cases involve the same point of protein-ligand attachment and the de livery of human insulin (or analogues or derivatives thereof) as the active species. Use of a acid-stabilized analog allows a stable, clear solution with ligand to be formulated at slightly acidic pH. Following subcutaneous injection, the pH is gradually adjusted towards neutral. As a result the ligand binds to and precipitates insulin in the subcutaneous tissue. The release of 10 insulin analog into the blood stream is then limited by the rate of redissolution of the precipi tate. Of particular advantage is the possibility of adjusting the amount of added ligand as well as the charge and affinity of the ligand. Variation of these parameters allows adjustment of the rate of dissolution following precipitation in the subcutis and hence the proportion of slow and fast acting analog in the formulationon. Hence formulations covering a wide range of re 15 lease rates may be prepared by this principle. The anions currently used in insulin formulations as allosteric ligands for the R-state hexam ers (notably chloride ion) bind only weakly to the His B ° anion site. The present invention, which is based on the discovery of suitable higher affinity ligands for these anion sites, pro vides ligands which are extended to modify timing via changes in hexamer solubility as out 20 lined above. Most ligand binding sites in proteins are highly asymmetric. Because the His B 10 Zn 2 + sites reside on the three-fold symmetry axis, these sites posses a symmetry that is unusual, but not unique. Several other proteins have highly symmetric ligand binding sites. The His" 1 a Zn2+ site consists of a tunnel or cavity with a triangular-shaped cross-section that 25 extends ~12 A from the surface of the hexamer down to the Hissio Zn 2 + ion. The diameter of the tunnel varies along its length and, depending on the nature of the ligand occupying the site, the opening can be capped over by the Asn1 3 and Phe
B
' side chains. The walls of the tunnel are made up of the side chains of the amino acid residues along one face each of the three a-helices. The side chains from each helix that make up the lining of the tunnel are 30 Phe" 1 , Asn1 3 , and Leu B6 . Therefore, except for the zinc ion, which is coordinated to three Hiss"o residues and is positioned at the bottom of the tunnel, the site is principally hydropho bic. Depending on the ligand structure, it may be possible for substituents on the ligand to make H-bonding interactions with Asn B3 and with the peptide linkage to Cys B7 . The present invention originates from a search for compounds with suitable binding proper 35 ties by using UV-visible and fluorescence based competition assays described herein which WO 2004/080480 PCT/DK2004/000158 12 are based on the displacement of chromophoric ligands from the R-state HisB 10 -Zn 2 site by the incoming ligand in question. These compounds will be referred to as "starter compounds" in the following. These assays are easily transformed into a high-throughput format capable of handling libraries constructed around hits from the initial search of compound databases. 5 These starter compounds provide the starting point for the task of constructing a chemical handle that allows for attachment of the positively charged fragment Frg2 (see below). Thus, from the structure-activity relationship (SAR) information obtained from the binding as 10 say(s) it will be apparent for those skilled in the art to modify the starter compounds in ques tion by introduction of a chemical group that will allow for coupling to a peptide containing e.g. one or more arginine or lysine residues. These chemical groups include carboxylic acid (amide bond formation with the peptide), carbaldehyde (reductive alkylation of the peptide), sulfonyl chloride (sulphonamide formation with the peptide) or the like. 15 The decision where and how to introduce this chemical group can be made in various ways. For example: From the SAR of a series of closely related starter compounds, a suitable posi tion in the starter compound can be identified and the chemical group can be attached to this position, optionally using a spacer group, using synthesis procedures known to those skilled 20 in the art. Alternatively, this chemical group can be attached (optionally using a spacer group using and synthesis procedures known to those skilled in the art) to a position on the starter compound remote from the Zn 2 "-binding functionality. 25 The invention thus provides pharmaceutical preparation comprising 1. Acid-stabilised insulin 2. Zinc ions 3. A zinc-binding ligand of the following general formula (I) 30 CGr-Lnk-Frg1-Frg2-X (I) wherein: 35 CGr is a chemical group which reversibly binds to a His
B
IO Zn 2 + site of an insulin hexamer; WO 2004/080480 PCT/DK2004/000158 13 Lnk is a linker selected from *a valence bond Sa chemical group G B of the formula -B 1
-B
2 -C(O)-, -B 1
-B
2
-SO
2 -, -B'-B 2
-CH
2 -, or -B 1 5 B 2 -NH-; wherein B 1 is a valence bond, -0-, -S-, or -NR 6 -,
B
2 is a valence bond, Cl-C 1 8 -alkylene, C 2
-C
18 -alkenylene, C 2
-C
1 8 -alkynylene, arylene, heteroarylene, -CI-ClV 8 -alkyl-aryl-, -C 2
-C
1 8 -alkenyl-aryl-, -C 2
-C
18 -alkynyl-aryl-, -C(=O) C1-Cj8-alkyI-C(=O) - , -C(=O)-Cj-Cj8-alkenyl-C(=O)-, -C(=O)-C,-Ci8-alkyl-O-C,-C18 alkyl-C(=O)-, -C(=O)- Cl-C 18 -alkyl-S-Cl-C 1 s-alkyl-C(=O)-, -C(=O)-C 1 -C18-alkyl-NR 6 -Cj 10 C, 8 -alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C(=O)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF 3 , -OCF 3 , -ORO, or -NR 6
R
7 and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR 6 , -C(O)H, OCOR 6 , -SO 2 , -CN, -CF 3 , OCF 3 , -NO 2 , -OR 6 , -NR 6
R
7 , C 1 -Ce 18 -alkyl, or Cs-C 18 -alkanoyl; 15 R 6 and R 7 are independently H, CI-C 4 -alkyl; Frgl is a fragment consisting of 0 to 5 neutral x- or p-amino acids Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from 20 amino or guanidino groups; and X is -OH, -NH 2 or a diamino group, or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or 25 mixture of optical isomers, including a racemic mixture, or any tautomeric forms. In one embodiment CGr is a chemical structure selected from the group consisting of car boxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imrni dazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia 30 zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids. In another embodiment CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5 35 mercaptotetrazoles, or 4-cyano-1,2,3-triazoles.
WO 2004/080480 PCT/DK2004/000158 14 In another embodiment CGr is vN A HN or HN, NB R
R
3 O R2 O R wherein 5 Xis=O,=Sor=NH Y is -S-, -0- or -NH
R
1 and R 4 are independently selected from hydrogen or Cl-C 6 -alkyl,
R
2 is hydrogen or Cl-C 6 -alkyl or aryl, R 1 and R 2 may optionally be combined to form a double 10 bond,
R
3 and R 5 are independently selected from hydrogen, halogen, aryl, C 1
-C
6 -alkyl, or -C(O)NR" R 12 , A and B are independently selected from Cl-C 6 -alkylene, arylene, aryl-C 1
-C
6 -alkyl-, aryl-C 2 15 C6-alkenyl- or heteroarylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 6 and the arylene or heteroarylene is optionally substituted with up to four substituents R 7 , R 8 , R, and R 1 0 , A and R 3 may be connected through one or two valence bonds, B and R 5 may be connected through one or two valence bonds, 20 R 6 is independently selected from halogen, -CN, -CF3, -OCF 3 , aryl, -COOH and -NH 2 ,
R
7 , R 8 , R 9 and R"' are independently selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2 CFa, -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 11 , -NR 11
R
12 , -SR 11 , 25 -NR"lS(O) 2
R
12 , -S(O) 2
NR"R
1 2 , -S(O)NR"R 12 , -S(O)R 11 , -S(O) 2
R
11 , -OS(O) 2
R
11 ,
-C(O)NR"R
12 , -OC(O)NR"R 1 2 , -NR 11
C(O)R
2 , -CH 2
C(O)NR"R
12
-OC-C
6 -alkyl-C(O)NR 1
"R
12 , -CH 2 OR", -CH 2 OC(O)R", -CH 2 NR"R 12 , -OC(O)R 1 , -OC1-C15-alkyl-C(O)OR", -OC,-C6-alkyl-OR", -SCj-C6-alkyl-C(O)OR"
-C
2
-C
6 -alkenyl-C(=O)OR 1 1 , -NR 1 1
-C(=O)-C
l
-C
6 -alkyl-C(=O) O R 1 , 30 -NR"-C(=O)-Cl-C 6 -alkenyl-C(=0)OR" 1 , -C(0)OR" 1 , C(O)R 1 1 , or -C 2
-C
6 -alkenyl
C(=O)R"
1 , =0, or -C 2
-C
6 -alkenyl-C(=O)-NR"R 12
,
WO 2004/080480 PCT/DK2004/000158 15
*C-C
6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, each of which may optionally be substi tuted with one or more substituents independently selected from R 13 * aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, 5 aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2 -Ce-alkynyl, heteroaryl, heteroaryl-C,
C
6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl, heteroaryl-C 2
-C
6 -alkynyl, or C 3
-C
6 cycloalkyl, of which each cyclic moiety may optionally be substituted with one or more substitu ents independently selected from R 14 10
R"
1 and R 2 are independently selected from hydrogen, OH, Cl-C20-alkyl, aryl-C-C6-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R'6; R 1 1 and R" when attached to the same 15 nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R'
13 is independently selected from halogen, -CN, -CF3, -OCF 3 , -OR", -C(O)OR" 1 , -NR"R 12, 20 and -C(O)NR"R 12 , Ri 4 is independently selected from halogen, -C(O)OR", -CH 2 C(O)OR", -CH 2
OR
1 1 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR", -NR"R1 2 , S(O) 2 R", aryl and Cl-C 6 -alkyl, 25 R s
'
5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OCl-C 6 -alkyl, -C(O)OC,-C6 alkyl, -COOH and -NH 2 , R" is independently selected from halogen, -C(O)OC,-C 6 -alkyl, -COOH, -CN, -CF3, -OCF 3 , NO 2 , -OH, -OCi-C6-alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, or any enantiomer, diastereomer, in 30 cluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically accept able acid or base. In another embodiment X is =0 or =S. In another embodiment X is =0. 35 In another embodiment X is =S.
WO 2004/080480 PCT/DK2004/000158 16 In another embodiment Y is -0- or -S-. In another embodiment Y is -0-. In another embodiment wherein Y is -S-. In another embodiment Crg is arylene optionally substituted with up to four substituents, R 7 , 5 R 8 , R 9 , and R 1 0 which may be the same or different. In another embodiment A is selected from ArG1 optionally substituted with up to four sub stituents, R 7 , R 8 , R, and R'O which may be the same or different. In another embodiment A is phenylene or naphtylene optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 o which may be the same or different. 10 In another embodiment A is R
R
9 or 7R R 7 R 8 RR In another embodiment A is phenylene. In another embodiment A is heteroarylene optionally substituted with up to four substituents,
R
7 , R 8 , R 9 , and R 1 0 which may be the same or different. 15 In another embodiment A is selected from Hetl optionally substituted with up to four sub stituents, R 7 , R 8 , R 9 , and R1 0 which may be the same or different. In another embodiment A is selected from Het2 optionally substituted with up to four sub stituents, R 7 , R 8 , R 9 , and R' 0 which may be the same or different. In another embodiment A is selected from Het3 optionally substituted with up to four sub 20 stituents, R 7 , R 8 , R 9 , and Rio which may be the same or different. In another embodiment A is selected from the group consisting of indolylene, benzofu ranylidene, quinolylene, furylene, thienylene, or pyrrolylene, wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 o which may be the same or different. 25 In another embodiment A is benzofuranylene optionally substituted with up to four substitu ents R 7 , R 8 , R 9 , and R 1 o which may be the same or different. In another embodiment A is
R
8 RB RRR Sor or 7 7 WO 2004/080480 PCT/DK2004/000158 17 In another embodiment A is carbazolylidene optionally substituted with up to four substitu ents R 7 , R 8 , R 9 , and Rio which may be the same or different. In another embodiment A is N
R
7 5 In another embodiment A is quinolylidene optionally substituted with up to four substituents
R
7 , R 8 , R 9 , and R 1 o which may be the same or different. In another embodiment A is 9 Nor or N R 8
R
7 R In another embodiment A is indolylene optionally substituted with up to four substituents R 7 , 10 R 8 , R 9 , and R 1 o which may be the same or different. In another embodiment A is R9 R R 9 R 8 R 9 R aR 9 R8 N NH NH orN 1R R R In another embodiment R 1 is hydrogen. In another embodiment R 2 is hydrogen. 15 In another embodiment R 1 and R 2 are combined to form a double bond. In another embodiment R 3 is Cl-C 6 -alkyl, halogen, or C(O)NR 6
R
17 . In another embodiment R 3 is Cl-CO-alkyl or C(O)NR 16 R 7 . In another embodiment R 3 is methyl. In another embodiment B is phenylene optionally substituted with up to four substituents, R 7 , 20 R 8 , R 9 , and R 10 which may be the same or different. In another embodiment R 4 is hydrogen. In another embodiment R s is hydrogen. In another embodiment R 6 is aryl.
WO 2004/080480 PCT/DK2004/000158 18 In another embodiment R 6 is phenyl. In another embodiment R 7 , R 8 , R 9 and R 1 0 are independently selected from *hydrogen, halogen, -NO 2 , -OR 1 1 , -NR"R 1 2 , -SR", -NR 1 lS(O) 2
R
12 , -S(O) 2
NR"R
2 , 5 -S(O)NR"R 12 , -S(O)R 1 , -S(O) 2
R
1 , -OS(0)2 R 11 , -NR"C(O)R 12 , -CH 2
OR
11 , CH20C(O)R 11 , -CH 2 NR"R 12 , -OC(O)R 1 1 , -OCl-C 6 -alkyl-C(O)OR" 1 , -OC1-C alkyl-C(O)NR 1
R
12 , -OCI-C 6 -alkyl-OR 11 , -SC 1
-C
6 -alkyl-C(O)OR 11 , -C 2
-C
6 -alkenyl
C(=O)OR
1 1 , -C(O)OR 1 1 , or -C 2
-C
6 -alkenyl-C(=O)R 1 , 10 * C
I
-C
6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted with one or more substituents independently selected from R 13 * aryl, aryloxy, aroyl, arylsulfanyl, aryl-C 1
-C
6 -alkoxy, aryl-C l
-C
6 -alkyl, aryl-C 2 C 6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, 15 wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 In another embodiment R 7 , R 8 , R 9 and R 1 0 are independently selected from 20 *hydrogen, halogen, -NO 2 , -OR 11 , -NR 11
R
12 , -SR 11 , -S(O) 2
R
11 , -OS(0)2 R 11 , CH 2
OC(O)R
11 , -OC(O)R 11 , -OCI-C 6 -alkyl-C(O)OR 1 , -OC1-C 6 -alkyl-OR 11 , -SC 1
-C
6 alkyl-C(O)OR 1 , -C(O)OR 1 , or -C 2
-C
6 -alkenyl-C(=O)R 11 , * C 1
-C
6 -alkyl or C 1
-C
6 -alkenyl which may each optionally be substituted with one or 25 more substituents independently selected from R 1 3 * aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-C 1
-C
6 -alkyl, heteroaryl, of which each of the cyclic moieties optionally may be substituted with one or more 30 substituents independently selected from R 14 In another embodiment R 7 , R 8 , R 9 and R' 0 are independently selected from WO 2004/080480 PCT/DK2004/000158 19 hydrogen, halogen, -NO 2 , -OR 11 , -NR"1R 12 , -SR 11 , -S(O) 2
R
11 , -OS(0)2 R 1 . CH 2
OC(O)R
11 , -OC(O)R 1 , -OC 1
-C
6 -alkyl-C(O)OR 11 , -OC 1
-C
6 -alky-OR 11, -Sc i-C6 alkyl-C(O)OR 1 , -C(O)OR 11 , or -C 2
-C
6 -alkenyl-C(=O)R" 1 , 5 *Cl-Ce-alkyl or Cl-C 6 - which may each optionally be substituted with one or more substituents independently selected from R 13 earyl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, 10 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 In another embodiment R 7 , R 8 , R 9 and R 1 0 are independently selected from * hydrogen, halogen, -OR" 1 , -OC1-C 6 -alkyl-C(O)OR 11 , or -C(O)OR 1 , 15 * Cl-C 6 -alkyl which may each optionally be substituted with one or more substituents independently selected from R 1 3 * aryl, aryloxy, aryl-Cl-C 6 -alkoxy, 20 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from Ri. In another embodiment R 7 , R 8 , R 9 and R 10 are independently selected from 25 *hydrogen, halogen, -OR 11 , -OC 1
-C
6 -alkyl-C(O)OR 1 , or -C(O)OR 11 , * Cl-C 6 -alkyl which may optionally be substituted with one or more substituents inde pendently selected from R 1 3 30 * phenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, wherein each of the cyclic moieties option ally may be substituted with one or more substituents independently selected from
R
14 In another embodiment R 1 1 and R 12 are independently selected from hydrogen, Cl-C 20 -alkyl, 35 aryl or aryl-Cl-C 6 -alkyl, wherein the alkyl groups may optionally be substituted with one or WO 2004/080480 PCT/DK2004/000158 20 more substituents independently selected from R 5 is , and the aryl groups may optionally be substituted one or more substituents independently selected from R'16; R" 11 and R' 2 when at tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms 5 selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds. In another embodiment R 1 and R1 2 are independently selected from hydrogen, Cl-C 2 0 -alkyl, aryl or aryl-Cl-C 6 -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 1 5 , and the aryl groups may optionally be 10 substituted one or more substituents independently selected from R1. In another embodiment R" and R 2 are independently selected from phenyl or phenyl-Cl-C 6 alkyl. In another embodiment R 1 " and R 12 are methyl. In another embodiment R 13 is independently selected from halogen, CF 3 , OR" or NR"R 12 . 15 In another embodiment R' 3 is independently selected from halogen or OR 1 ". In another embodiment R1 3 is OR". In another embodiment R1 4 is independently selected from halogen, -C(O)OR", -CN, -CF 3 , OR", S(O) 2 R", and Cl-C6-alkyl. In another embodiment R1 4 is independently selected from halogen, -C(O)OR", or -OR". 20 In another embodiment R1 s is independently selected from halogen, -CN, -CF 3 , -C(O)OCI-C 6 alkyl,and -COOH. In another embodiment R 15 is independently selected from halogen or -C(O)OCI-C 6 -alkyl. In another embodiment R'" is independently selected from halogen, -C(O)OC-C 6 -alkyl, -COOH, -NO 2 , -OC,-C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl. 25 In another embodiment R 1 " is independently selected from halogen, -C(O)OCI-C 6 -alkyl, -COOH, -NO 2 , or Cl-C 6 -alkyl. In another embodiment CGr is R ~ 20 F or N E or NG 18N R H H O 30 wherein R1 9 is hydrogen or Cl-C 6 -alkyl,
R
2 0 is hydrogen or C,-C 6 -alkyl, WO 2004/080480 PCT/DK2004/000158 21 D and F are a valence bond or Cl-C 6 -alkylene optionally substituted with one or more sub stituents independently selected from R 72 , 5 R 72 is independently selected from hydroxy, C 1
-C
6 -alkyl, or aryl, E is C 1
-C
6 -alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is option ally substituted with up to three substituents R 21 , R 22 and R 23 , G is Cl-C 6 -alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is op 10 tionally substituted with up to three substituents R 24 , R 2 and R 2 , R", R 18 ', R 2 1 , R 2 2 , R 23 , R 24 , R 2 5 and R 2 6 are independently selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , 15 -OCF 2
CHF
2 , -S(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 7 , -NR 27
R
28 , -SR 27 , -NR 27
S(O)
2 R",
-S(O)
2
NR
27
R
28 , -S(O)NR 27
R
28 , -S(O)R 27 , -S(O) 2
R
27 , -C(O)NR 27
R
2 , -OC(O)NR 27
R
28
-NR
2 7
C(O)R
8 , -NR 27
C(O)OR
2 8 , -CH 2
C(O)NR
7
R
28 , -OCH 2
C(O)NR
27
R
2 8 , -CH 2 0R? 7 ,
-CH
2
NR
27
R
28 , -OC(O)R 27 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC 1
-C
6 -alkyl-C(O)OR 27 , -C 2
-C
6 alkenyl-C(=O)OR 27 , -NR 27 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 2 7 , -NR 27
-C(=O)-C
1
-C
6 20 alkenyl-C(=O)OR 27 , -C(=O)NR 27
-C
1 -Ce-alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 27 ,or
-C(O)OR
2 7 * CI-Co-alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, 25 which may optionally be substituted with one or more substituents independently se lected from R 29 * aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-C 1
-C
6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2
-C
6 30 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 , 35 WO 2004/080480 PCT/DK2004/000158 22
R
27 and R 2a are independently selected from hydrogen, C-C 6 -alkyl, aryl-C 1
-C
6 -alkyl or aryl, or
R
2 7 and R 2 8 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two 5 double bonds,
R
29 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 27 , and -NR 2 7
R
2 8 ,
R
3 0 is independently selected from halogen, -C(O)OR 2 7 , -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 2 7 10 -NR 27
R
2 8 and Cl-Co-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. In another embodiment D is a valence bond. In another embodiment D is Ci-C 6 -alkylene optionally substituted with one or more hydroxy, Cl-C 6 -alkyl, or aryl. 15 In another embodiment E is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents independently selected from R 21 , R' and
R
23 In another embodiment E is arylene optionally substituted with up to three substituents inde pendently selected from R 21 , R 22 and R 23 20 In another embodiment E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 . In another embodiment E is phenylene optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 In another embodiment CGr is R22 0 N J19
R
21 N"R 25 H In another embodiment R 21 , R2 and R 23 are independently selected from *hydrogen, halogen, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -SCF 3 , 30 NO 2 , -OR 27 , -NR 27
R
2 8 , -SR 27 , -C(O)NR 27
R
2 8 , -OC(O)NR 27
R
2 8 , -NR 27
C(O)R
2 8 ,
-NR
27
C(O)OR
2 8 , -CH 2
C(O)NR
27
R
28 , -OCH 2
C(O)NR
27
R
28 , -CH 2 0R 27 , -CH 2
NR
27
R
28 ,
-OC(O)R
27 , -OC,-C 6 -alkyl-C(O)OR 2 7 , -SCI-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl- WO 2004/080480 PCT/DK2004/000158 23
C(=O)OR
27 , -NR 27
-C(=O)-C
1
-C
6 -alkyl-C(=O)OR 27 , -NR27-C(=o)-C1.C_ alkenyl-C(=O)
O
R
27 -, -C(=O)NR 2 7 -Cl-C-alkyl-C(=O)OR 2 7 , -Ci -C 6 -alkyl-C(=O)OR1 7 , or
-C(O)OR
27 , 5 * Cl-C6-alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 2 9 10 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-CB-alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 Ce-alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu 15 ents selected from R 30 In another embodiment R 2 1 , R 2 and R 23 are independently selected from * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 2 7
R
2 a , -SR 27 , -NR 2 7
C(O)R
28 , -NR 27
C(O)OR
28 , 20 -OC(O)R 27 , -OCl-C 6 -alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)
O
R
27 , -C(=O)N R 27
-C
1
-C
6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 2 7 , or
-C(O)OR
27 * Cl-C6-alkyl optionally substituted with one or more substituents independently se 25 lected from R2 * aryl, aryloxy, aroyl, aryl-Cl-Ce-alkoxy, aryl-C 1
-C
6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl, 30 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . In another embodiment R 21 , R' and R 23 are independently selected from WO 2004/080480 PCT/DK2004/000158 24 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
2 8 , -SR 2 7 , -NR 27
C(O)R
2 a , -NR 27
C(O)CR
2 , -OC(O)R, -OCs-C 6 -alkyl-C(0)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alcenyl
C(=O)OR
27 , -C(=O)NR 27 -Cj-C 6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(-=O)OR 27 , or
-C(O)OR
27 , 5 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R2 * aryl, aryloxy, aroyl, aryl-C 1 -C6-alkoxy, aryl-C l
-C
6 -alkyl, heteroaryl, heteroaryl-C,-C6 10 alkyl of which the cyclic moieties optionally may be substituted with one or more sub stitu ents selected from R 3 . In another embodiment R 2 1 , R 22 and R 2 3 are independently selected from 15 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
2 8 , -NR 27
C(O)OR
28 ,
-OC(O)R
2 7 , -OC,-C 6 -alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 2 7 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 2 7 -Cl-C6-alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 27 , or
-C(O)OR
7 , 20 *methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 *ArG1, ArG1-O-, ArG1-C(O)-, ArG1-Cl-C 6 -alkoxy, ArG1-C 1
-C
6 -alkyl, Het3, Het3-Cl
C
6 -alkyl 25 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30 In another embodiment R 21 , R 22 and R 23 are independently selected from *hydrogen, halogen, -OCF 3 , -OR 2 7 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
28 , -NR 2 7
C(O)OR
28 , 30 -OC(O)R 27 , -OC-C 6 -alkyl-C(O)OR 27 , -SC,-CB-alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 2 -Cl-C 6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 27 , or
-C(O)OR
2 7 , * C-C 6 C-alkyl optionally substituted with one or more substituents independently se 35 lected from R 29 WO 2004/080480 PCT/DK2004/000158 25 Sphenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, phenyl-Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . 5 In another embodiment R 19 is hydrogen or methyl. In another embodiment R 19 is hydrogen. In another embodiment R 27 is Hydrogen, Cl-C 6 -alkyl or aryl. In another embodiment R 27 is hydrogen or Cl-C 6 -alkyl. In another embodiment R 28 is hydrogen or C 1
-C
6 -alkyl. 10 In another embodiment F is a valence bond. In another embodiment F is C 1
-C
6 -alkylene optionally substituted with one or more hydroxy, Cl-C 6 -alkyl, or aryl. In another embodiment G is Cl-Ce-alkylene or arylene, wherein the arylene is optionally sub stituted with up to three substituents R 24 , R 2 5 and R 26 . 15 In another embodiment G is Cl-Co-alkylene or ArG1, wherein the arylene is optionally substi tuted with up to three substituents R 24 , R 25 and R 26 . In another embodiment G is C 1
-C
6 -alkylene. In another embodiment G is phenylene optionally substituted with up to three substituents
R
24 , R 2s and R 26 . 20 In another embodiment R 24 , R 25 and R 26 are independently selected from Hydrogen, halogen, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -SCF 3 , NO 2 , -OR 27 , -NR 27
R
28 , -SR 27 , -C(O)NR 27
R
28 , -OC(O)NR 2 7
R
28 , -NR 27
C(O)R
28 ,
-NR
27
C(O)OR
28 , -CH 2
C(O)NR
27
R
28 , -OCH 2
C(O)NR
27
R
28 , -CH 2 0R 27 , -CH 2
NR
27
R
28 , 25 -OC(O)R 2 7 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC 1
-C
6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -NR 27 -C(=O)-C -C 6 -alkyl-C(=O)OR 27 , -NR 27
-C(=O)-C-C
6 alkenyl-C(=O)OR 27 -, -C(=O)NR 27
-C
6 -C-alkyl-C(=O)OR 27 , -Cj-C 6 -alkyl-C(=O)OR 27 , or
-C(O)OR
27 30 * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R4 WO 2004/080480 PCT/DK2004/000158 26 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-C 1
-C
6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroary-C 1
-C
6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . In another embodiment R 24 , R 25 and R 26 are independently selected from 10 *hydrogen, halogen, -OCF 3 , -OR 2 , -NR 27
R
28 , -SR 2 7 , -NR 27
C(O)R
8 , -NR 27
C(O)OR
28
-OC(O)R
27 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 2 7 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 27 Cl -C 6 -alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 27 , or -C(O)OR", 15 * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 2 9 20 e aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C-C 6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu 25 ents selected from R 3 . In another embodiment R 2 4 , R 25 and R 26 are independently selected from * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
28 , -NR 27
C(O)OR
28 , 30 -OC(O)R 27 , -OCj-C 6 -alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 2 7 Cl -C 6 -alkyl-C(=O)OR 2 7 , -C 1
-C
6 -alkyl-C(=O)OR 27 , or
-C(O)OR
7 , * C 1
-C
6 -alkyl optionally substituted with one or more substituents independently se 35 lected from R 2 9 WO 2004/080480 PCT/DK2004/000158 27 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-CB alkyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 ". In another embodiment R 24 , R 25 and R 26 are independently selected from 10 *.hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 2T
C(O)R
28 , -NR 27
C(O)OR
8 ,
-OC(O)R
2 7 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SCI-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 27 C -C 6 -alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 7 , or
-C(O)OR
27 15 e methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 *ArG1, ArG1-O-, ArGI-C(O)-, ArG1-Cl-C 6 -alkoxy, ArG1-Cl-C 6 -alkyl, Het3, Het3-C,
C
6 -alkyl 20 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . In another embodiment R 24 , R 25 and R 2 6 are independently selected from * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
2 8 , -NR 27
C(O)OR
28 , 25 -OC(O)R 27 , -OCl-C 6 -alkyl-C(O)OR 2 7 , -SCI-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)N R 27 -Cl -C 6 -alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 2 7 , or
-C(O)OR
27 * methyl, ethyl propyl optionally substituted with one or more substituents independ 30 ently selected from R 29 *ArG1, ArG1-O-, ArG1-C(O)-, ArG1-Cl-C 6 -alkoxy, ArG1-Cl-C 6 -alkyl, Het3, Het3-C 1 C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substitu 35 ents selected from R 30 WO 2004/080480 PCT/DK2004/000158 28 In another embodiment R 24 , R 25 and R 26 are independently selected from *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
28 , -NR27C(O)OR 28 ,
-OC(O)R
27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl 5 C(=O)OR 27 , -C(=O)N R 27
-C
1
-C
6 -alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 27 , or
-C(O)OR
27 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 10 * ArG1, ArG1 -O-, ArG 1-Cl-C 6 -alkoxy, ArG 1 -Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30 . In another embodiment R 2 0 is hydrogen or methyl. 15 In another embodiment R 20 is hydrogen. In another embodiment R 27 is hydrogen, CI-C 6 -alkyl or aryl. In another embodiment R 27 is hydrogen or Cl-C 6 -alkyl or ArG1. In another embodiment R 2 7 is hydrogen or Cl-C 6 -alkyl. In another embodiment R 2 8 is hydrogen or Cl-C 6 -alkyl. 20 In another embodiment R 17 and R 18 are independently selected from * hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 27 , -NR 27
R
28 , -SR 27 , -S(O)R 2 7
-S(O)
2
R
27 , -C(O)NR 27
R
2 8 , -CH 2 0R 27 , -OC(O)R 27 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC 1
-C
6 alkyl-C(O)OR 27 , or -C(O)OR 27 25 * C,-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, optionally substituted with one or more substituents independently selected from R 2 9 earyl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C6 30 alkyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . In another embodiment R 1 7 and R' are independently selected from 35 WO 2004/080480 PCT/DK2004/000158 29 *hydrogen, halogen, -CN, -CFa, -NO 2 , -OR 27 , -NR 2 7
R
28 , or -C(O)OR 27 * C,-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 29 5 e aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl, of which the cyclic moieties optionally may be substituted with one or more substitu 10 ents selected from R 30 In another embodiment R 17 and R' 8 are independently selected from * hydrogen, halogen, -CN, -CF3, -NO 2 , -OR 27 , -NR 2 7
R
28 , or -C(O)OR 27 * methyl, ethyl propyl optionally substituted with one or more substituents independ 15 ently selected from R 29 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . 20 In another embodiment R 17 and R 18 are independently selected from * hydrogen, halogen, -CN, -CF3, -NO 2 , -OR 27 , -NR 27
R
28 , or -C(O)OR 27 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 eArG1, ArGI-O-, ArG1-C(O)-, ArGl-Cl-C 6 -alkoxy, ArGl-Cl-C 6 -alkyl, Het3, Het3-C 1 25 C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30 . In another embodiment R" 7 and R" are independently selected from * hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 27
R
28 , or -C(O)OR 27 30 * C 1
-C
6 -alkyl optionally substituted with one or more substituents independently se lected from R 9 * phenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, phenyl-Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . 35 In another embodiment R 27 is hydrogen or Cl-C 6 -alkyl.
WO 2004/080480 PCT/DK2004/000158 30 In another embodiment R 27 is hydrogen, methyl or ethyl. In another embodiment R 28 is hydrogen or Cl-C 6 -alkyl. In another embodiment R 2 8 is hydrogen, methyl or ethyl. In another embodiment R 72 is -OH or phenyl. 5 In another embodiment CGr is N) N H In another embodiment CGr is of the form H-I-J wherein H is 10 0 0 OH 0 OH HO or HO or HO HHOor N H 15 wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 I is selected from 20 * a valence bond, * -CH 2
N(R
32 )- or -SO 2
N(R
33 )-, -ZL-N--n * wherein Z' is S(O) 2 or CH 2 , Z 2 is -NH-, -O-or -S-, and n is 1 or 2, J is 25 * Cl-C 6 -alkylene, C 2
-C
6 -alkenylene or C 2
-C
6 -alkynylene, which may each optionally be substituted with one or more substituents selected from R3, *Arylene, -aryloxy-, arylene-oxycarbonyl-, -aroyl, arylene-Ci-C 6 -alkoxy-, ary lene-Cl-C 6 -alkylene, arylene-C 2
-C
6 -alkenylene, arylene-C 2
-C
6 -alkynylene, heteroary lene, heteroarylene-Cl-C-alkylene-, heteroarylene-C 2
-C
6 -alkenylene or heteroary 30 lene-C 2
-C
6 -alkynylene, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R 37
,
WO 2004/080480 PCT/DK2004/000158 31
R
31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CFa, -OCF 3 ,
-OCHF
2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -S(O) 2
CF
3 , -SCF 3 , -NO 2 , -ORa s , -C(O)R 3 , -NR 5
R
36 , -SR 35 ,
-NR
35
S(O)
2
R
3 6 , -S(O) 2
NR
3 5
R
36 , -S(O)NR 35
R
36 , -S(O)R 35 , -S(O) 2
R
35 , -C(O)NR 3 5 sR 36 , 5 -OC(O)NR R 35
R
36 , -NR 35 C(O)R36, -CH 2
C(O)NR
35
R
36 , -OCH 2
C(O)NR
35 sR, -CH 2 0R 3 5 ,
-CH
2
NR
35 R36, -OC(O)R 35 , -OCl-C 6 -alkyl-C(O)OR 35 , -SC 1
-C
6 -alkyl-C(O)OR 3 5
-C
2
-C
6 -alkenyl
C(=O)OR
35 , -NR35-C(=O)-C-C 6 -alkyl-C(=O)OR", -N R 3
-C(=O)-C
1
-C
6 -alkenyl-C(=O)O R-,
C
1 -Ce-alkyl, Cl-Ce-alkanoyl or -C(O)OR 3 5 10 R 32 and R3 are independently selected from hydrogen, Cl-C6-alkyl or C 1
-C
6 -alkanoyl,
R
34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 3 5 , and -NR 3 5R36,
R
35 and R 36 are independently selected from hydrogen, C 1
-C
6 -alkyl, aryl-C 1
-C
6 -alkyl or aryl, or 15 R 35 and R 3 6 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 20 R 37 is independently selected from halogen, -C(O)OR 35 , -C(O)H, -CN, -CF3, -OCF 3 , -NO 2 , OR 35 , -NR 35
R
3 6 , C 1
-C
6 -alkyl or Cl-C 6 -alkanoyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. 25 In another embodiment H is O 0 HO or HO orI HO HO In another embodiment H is WO 2004/080480 PCT/DK2004/000158 32 0 HO HO In another embodiment H is 0 HO O " HO In another embodiment I is a valence bond, -CH 2
N(R
32 )-, or -SO 2
N(R
3 3 )-. 5 In another embodiment I is a valence bond. In another embodiment J is * Cl-C 6 -alkylene, C 2
-C
6 -alkenylene or C 2 -CB-alkynylene, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35
R
36 10 *arylene, or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 In another embodiment J is * arylene or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 . 15 In another embodiment J is e ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 . In another embodiment J is Sphenylene or naphthylene optionally substituted with one or more substitu 20 ents independently selected from R 37 . In another embodiment R 32 and R 33 are independently selected from hydrogen or C 1
-C
6 -alkyl. In another embodiment R3 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -OR 35 ,
-C(O)R
35 , -NR 35 sR 36 , -SR 35 , -C(O)NR 3 5
R
36 , -OC(O)NR 3 5 sR 36 , -NR 35
C(O)R
36 , -OC(O)R 3 1, -OCl
C
6 -alkyl-C(O)OR 3 5 , -SC,-C 6 -alkyl-C(O)OR 35 or -C(O)OR 5 . 25 In another embodiment R3 is hydrogen, halogen, -CF3, -NO 2 , -OR 35 , -NR R 36 , -SR 35 ,
-NR
35 C(O)R36, or -C(O)OR 3 5 . In another embodiment R3 is hydrogen, halogen, -CF 3 , -NO 2 , -OR", -NRaR 3 6, or
-NR
35
C(O)R
3 6 WO 2004/080480 PCT/DK2004/000158 33 In another embodiment R34 is hydrogen, halogen, or -OR 35 . In another embodiment R 35 and R3 are independently selected from hydrogen, CI-C 6 -alkyl, or aryl. In another embodiment R 35 and R" are independently selected from hydrogen or Cl-C 6 -alkyl. 5 In another embodiment R 37 is halogen, -C(O)OR 3 5 , -CN, -CF 3 , -OR 35 , -NR 3 sR 36 , C,-C 6 -alkyl or Cl-C 6 -alkanoyl. In another embodiment R" 7 is halogen, -C(O)OR 35 , -OR 35 , -NR 3 5
R
3 , C 1
-C
6 -alkyl or Ci-C alkanoyl. In another embodiment R 37 is halogen, -C(O)OR 3 5 or -OR". 10 In another embodiment CGr is HN N KM M N=N T wherein K is a valence bond, Cl-C 6 -alkylene, -NH-C(=O)-U-, -Cj-C 8 -alkyl-S-, -C 1
-C
6 -alkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any C 1 -Co-alkyl moiety is optionally substituted with R 3 5 , 15 U is a valence bond, Cl-C-alkenylene, -C 1 -Co-alkyl-O- or Cl-C 6 -alkylene wherein any Cj Co-alkyl moiety is optionally substituted with C 1
-C
6 -alkyl,
R
3 " is Cl-Co-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one 20 or more substituents independently selected from R 9 ,
R
3 9 is independently selected from halogen, cyano, nitro, amino, M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are 25 optionally substituted with one or more substituents independently selected from R 4 0
R
40 is selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 41 , -NR 41
R
42 , -SR 41 , 30 -NR 4 1
S(O)
2
R
42 , -S(O) 2
NR
4
R
42 , -S(O)NR 41
R
42 , -S(O)R 41 , -S(O) 2
R
41 , -OS(O) 2
R
41 ,
-C(O)NR
41
R
42 , -OC(O)NR 4 1
R
42 , -NR 4 1
C(O)R
42 , -CH 2
C(O)NR
41
R
42 , -OC 1
-C
6 alkyl-C(O)NR 41
R
42 , -CH 2 0R 4 1 , -CH 2
OC(O)R
41 , -CH 2
NR
41
R
42 , -OC(O)R 41 , -OC 1
-C
6 alkyl-C(O)OR 41 , -OCl-C 6 -alkyl-OR 4 ', -S-Cl-Co-alkyl-C(O)OR 4 1 , -C 2
-C
6 -alkenyl- WO 2004/080480 PCT/DK2004/000158 34
C(=O)OR
4 1 , -NR 41 -C(=O)-C -C6-alkyl-C(=O)OR 41 , -N R 41 -C(=0)-C 1
-C
6 alkenyl-C(=O)
O
R
41 , -C(O)OR 4 1 , -C 2
-C
6 -alkenyl-C(=O)R 41 , =0, -NH-C(=O)-O-CI
C
6 -alkyl, or -NH-C(=O)-C(=O)-O-C 1
-C
6 -alkyl, 5 * C 1
-C
6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R 43 *aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C 1 10 C 6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl or heteroaryl-C 2
-C
6 -alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from
R,
44
R
41 and R 42 are independently selected from hydrogen, -OH, C 1
-C
6 -alkyl, C-C 6 -alkenyl, aryl 15 C 1
-C
6 -alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substi tuted with one or more substituents independently selected from R 46 ; R 4 1 and R 42 when at tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms 20 selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R
43 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 41 , and -NR 4 1
R
42 25 R" 44 is independently selected from halogen, -C(O)OR 41 , -CH 2
C(O)OR
4 1 , -CH 2 0R 4 1 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 4 1
R
4 2 and Cl-C 6 -alkyl,
R
4 5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -O-Cl-C 6 -alkyl, -C(O)-O-Cl CB-alkyl, -COOH and -NH 2 ,
R
4 6 is independently selected from halogen, -C(O)OC-Co-alkyl, -COOH, -CN, -CF 3 , -OCF 3 , 30 NO 2 , -OH, -OCI-C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, Q is a valence bond, Cl-C 6 -alkylene, -Cl-C 6 -alkyl-O-, -C 1
-C
6 -alkyl-NH-, -NH-C-C 6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-Cl-C 6 -alkyl, -C(=O)-, or -Cl-C 6 -alkyl-C(=O)-N(R 4 1)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected 35 from R 4 , WO 2004/080480 PCT/DK2004/000158 35
R
4 7 and R 48 are independently selected from hydrogen, Cl-C 6 -alkyl, aryl optionally substituted with one or more R 49 , 5 R 49 is independently selected from halogen and -COOH, T is * Cl-C 6 -alkylene, C 2 -C-alkenylene , C 2
-C
6 -alkynylene, -Cl-C-alkyloxy-carbonyl, 10 wherein the alkylene, alkenylene and alkynylene moieties are optionally substituted with one or more substituents independently selected from R 5 0 , * arylene, -aryloxy-, -aryloxy-carbonyl-, arylene-Cl -C 6 -alkylene, -aroyl-, arylene-C 1 C 6 -alkoxy-, arylene-C2-C6-alkenylene, arylene-C2-C 6 -alkynylene, heteroarylene, het eroarylene-C-C 6 -alkylene, heteroarylene-C 2
-C
6 -alkenylene, heteroarylene-C 2 15 C 6 -alkynylene, wherein any alkylene, alkenylene , alkynylene, arylene and heteroarylene moiety is optionally substituted with one or more substituents independently selected from R
°
, 20 R s 50 is Cl-C 6 -alkyl, C 1
-C
6 -alkoxy, aryl, aryloxy, aryl-Cl-C 6 -alkoxy, -C(=O)-NH-Cj-C 6 -alkyl-aryl, heteroaryl, heteroaryl-Cl-C 6 -alkoxy, -C 1
-C
6 -alkyl-COOH, -O-Cl-C 6 -alkyl-COOH, -S(O) 2
R
51 ,
-C
2
-C
6 -alkenyl-COOH, -OR 5 1 , -NO 2 , halogen, -COOH, -CF 3 , -CN, =0, -N(R 5
'R
5 2 ), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R " , 25 R 51 and R 5 2 are independently selected from hydrogen and Cl-C 6 -alkyl,
R
53 is independently selected from Cl-C 6 -alkyl, Cl-C 6 -alkoxy, -C 1
-C
6 -alkyl-COOH, -C 2 C 6 -alkenyl-COOH, -OR 51 , -NO 2 , halogen, -COOH, -CF 3 , -CN, or -N(R 51
R
52 ), or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt 30 thereof with a pharmaceutically acceptable acid or base. In another embodiment K is a valence bond, Cl-C 6 -alkylene, -NH-C(=O)-U-, -C 1
-C
6 -alkyl-S-, -Cl-C 6 -alkyl-O-, or -C(=O)-, wherein any Cl-C 6 -alkyl moiety is optionally substituted with R 3 8 . In another embodiment K is a valence bond, Cl-C 6 -alkylene, -NH-C(=O)-U-, -Cl-C 6 -alkyl-S-, or -C 1
-C
6 -alkyl-O, wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R 38 WO 2004/080480 PCT/DK2004/000158 36 In another embodiment K is a valence bond, Cl-C 6 -alkylene, or -NH-C(=O)-U, wherein any
C
1
-C
6 -alkyl moiety is optionally substituted with R 38 . In another embodiment K is a valence bond or Cl-C 6 -alkylene, wherein any Cl-C 6 -alkyl moi ety is optionally substituted with R3. 5 In another embodiment K is a valence bond or -NH-C(=O)-U. In another embodiment K is a valence bond. In another embodiment U is a valence bond or -Cl-C 6 -alkyl-O-. In another embodiment U is a valence bond In another embodiment M is arylene or heteroarylene, wherein the arylene or heteroarylene 10 moieties are optionally substituted with one or more substituents independently selected from
R
40 In another embodiment M is ArG1 or Hetl, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 4 0 . In another embodiment M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are 15 optionally substituted with one or more substituents independently selected from R 40 In another embodiment M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 . In another embodiment M is phenylene optionally substituted with one or more substituents independently selected from R 40 . 20 In another embodiment M is indolylene optionally substituted with one or more substituents independently selected from R 40 In another embodiment M is 40 "" N In another embodiment M is carbazolylene optionally substituted with one or more substitu 25 ents independently selected from R 40 . In another embodiment M is e
R
40 In another embodiment R 4° is selected from WO 2004/080480 PCT/DK2004/000158 37 Hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 41
R
42 , -SR 41,
-S(O)
2
R
41 ,
-NR
4 1
C(O)R
42 , -OC-C 6 -alkyl-C(O)NR 4
'R
42 , -C 2
-C
6 -alkenyl-C(=O)OR 1 , -C(O)OR 4 1 =0, -NH-C(=0)-O-Cl-C-alkyl, or -NH-C(=O)-C(=O)-O-Cj-C 6 -alkyl, 5 C 1
-C
6 -alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 , * aryl, aryloxy, aryl-C0 1 -C-alkoxy, aryl-C-C 6 -alkyl, aryl-C2-C 6 -alkenyl, heteroaryl, het eroaryl-Cl-C 6 -alkyl, or heteroaryl-C2-C 6 -alkenyl, wherein the cyclic moieties optionally 10 may be substituted with one or more substituents selected from R". In another embodiment R 4 0 is selected from e hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 41
R
42 , -SR 41 , -S(0) 2
R
41 , -N R 41
'
C (
O)R
42 , -OC-C 6 -alkyl-C(O)N R 41
R
42 , -C 2 -C6-alkenyl-C(=O)O R 41 , -C(O)OR 41 , =0, -NH-C(=0)-O-C-C 6 -alkyl, or -NH-C(=O)-C(=O)-O-C-C 6 -alkyl, 15 Cl-C 6 -alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 , * ArG1, ArG1-O-, ArG1-C01-C 6 -alkoxy, ArG1-Cl-C 6 -alkyl, ArGl-C 2
-C
6 -alkenyl, Het3, 20 Het3-C-C 6 -alkyl, or Het3-C 2 -C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R 44 ". In another embodiment R 40 is selected from *hydrogen, halogen, -CF 3 , -NO 2 , -OR 41 , -NR 4 1
R
42 , -C(O)OR 41 , =0, or-NR 4 1
C(O)R
42 * C 1 -Co 6 -alkyl, 25 * ArG1. In another embodiment R 40 is selected from * Halogen, -NO 2 , -OR 41 , -NR 4 1
R
42 , -C(O)OR 4 1 , or -NR 41
C(O)R
42 , * Methyl, * Phenyl. 30 In another embodiment R 41 and R 42 are independently selected from hydrogen, C-C 6 -alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH. In another embodiment R 41 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or COOH.
WO 2004/080480 PCT/DK2004/000158 38 In another embodiment Q is a valence bond, C 1
-C
6 -alkylene, -Cl-C 6 -alkyl-O-, -Cl
C
6 -alkyl-NH-, -NH-C 1
-C
6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-Cl-C 6 -alkyl, -C(=O)-, or -Cl C6-alkyl-C(=O)-N(R 4 7 )- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R. 5 In another embodiment Q is a valence bond, -CH 2 -, -CH 2
-CH
2 -, -CH 2 -O-, -CH 2
-CH
2 -O-,
-CH
2 -NH-, -CH 2
-CH
2 -NH-, -NH-CH 2 -, -NH-CH 2
-CH
2 -, -NH-C(=O)-, -C(=O)-NH-, -O-CH 2 -,
-O-CH
2
-CH
2 -, or -C(=O)-. In another embodiment R 47 and R4 are independently selected from hydrogen, methyl and phenyl. 10 In another embodiment T is * C-C 6 -alkylene optionally substituted with one or more substituents independently selected from R 50 , *arylene, arylene-C 1
-C
6 -alkylene, heteroarylene, wherein the alkylene, arylene and heteroarylene moieties are optionally substituted with one or more substituents inde 15 pendently selected from R 5 0 . In another embodiment T is * Cl-C 6 -alkylene optionally substituted with one or more substituents independently selected from R5, *ArG1, ArGl-Cl-C 6 -alkylene, Het3, wherein the alkyl, aryl and heteroaryl moieties are 20 optionally substituted with one or more substituents independently selected from Rs. In another embodiment T is * Cl-C 6 -alkylene, optionally substituted with one or more substituents independently selected from R °, 25 e phenylene, phenylene-Cl-C 6 -alkylene, wherein the alkylene and phenylene moieties are optionally substituted with one or more substituents independently selected from
R
50 . In another embodiment R 5 o is C 1
-C
6 -alkyl, C 1
-C
6 -alkoxy, aryl, aryloxy, aryl-C 1 -C6-alkoxy, 30 -C(=O)-NH-C-C 6 -alkyl-aryl, heteroaryl, -C 1 -Co-alkyl-COOH, -O-Cl-C 6 -alkyl-COOH, -S(O) 2
R
51 ,
-C
2
-C
6 -alkenyl-COOH, -OR 51 , -NO 2 , halogen, -COOH, -CF 3 , -CN, =0, -N(Rs 5 'Rs 5 2 ), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R 5 3 . In another embodiment R 50 is Ci-C 6 -alkyl, C l
-C
6 -alkoxy, aryl, aryloxy, aryl-Cl-C 6 -alkoxy ,
-OR
51 , -NO 2 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with 35 one or more R 53
.
WO 2004/080480 PCT/DK2004/000158 39 In another embodiment R 5 is C 1
-C
6 -alkyl, aryloxy, aryl-Cl-C 5 -alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 3 . In another embodiment R s 50 is Cl-C 6 -alkyl, ArG1-O-, ArG1-Cl-C 6 -alkoxy , -OR", halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 . 5 In another embodiment R 5 is phenyl, methyl or ethyl. In another embodiment R 5 is methyl or ethyl. In another embodiment R 51 is methyl. In another embodiment R 53 is Cl-C 6 -alkyl, Cl-C 6 -alkoxy, -OR 51 , halogen,or -CF 3 . 10 In another embodiment CGr is N N HN ~ / S wherein V is C 1
-C
6 -alkylene, arylene, heteroarylene, arylene-Cl.6-alkylene or arylene-C2- 6 15 alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R5, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 5 , R5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 , 20 R 55 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 5 6 , -NR5R 5 7 , -SR 56 ,
-NR
5 6
S(O)
2
R
57 , -S(O) 2
NR
56
R
57 , -S(O)NRS 6
R
57 , -S(O)R 6 , -S(O) 2
R
56 , -OS(O) 2 R-,
-C(O)NR
56
R
57 , -OC(O)NRR 5 7 , -NR 56
C(O)R
57 , -CH 2
C(O)NR
56
R
57 , -OC-C6 25 alkyl-C(O)NR 6
R
57 , -CH 2
OR
s 56 , -CH 2 0C(O)R 56 , -CH 2
NR
5 Rs 57 , -OC(O)R 56 , -OC 1 -Ca alkyl-C(O)OR 56 , -OCI-C 6 -alkyl-OR", -SCi-C 6 -alkyl-C(O)OR 5 6 , -C 2
-C
6 -alkenyl C(=O)OR 5 , -NR-C(=O)-Cl-C 6 -alkyl-C(=O)OR 5 6 , -NR 5 1-C(=O)-Cl-C6 alkenyl-C(=O)OR 56 , -C(O)OR 56 , or -C 2 -Co-alkenyl-C(=O)R
M
, 30 * C l
-C
6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents selected from IR
"
,
WO 2004/080480 PCT/DK2004/000158 40 * aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C Cr-alkyl, heteroaryl-C 2
-C
6 -alkenyl or heteroaryl-C 2
-C
6 -alkynyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R5 9 ,
R
6 and R5 7 are independently selected from hydrogen, OH, CF 3 , C 1
-C
1 2 -alkyl, aryl-C-C 6 alkyl, -C(=O)-Cl-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with 10 one or more substituents independently selected from R 60 , and the aryl groups may option ally be substituted with one or more substituents independently selected from R"'; R 56 and
R
57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two 15 double bonds, R5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 56 , and -NR 5 6
R
57 ,
R
59 is independently selected from halogen, -C(O)OR 6 , -CH 2
C(O)OR
56 , -CH 2 OR 6 , -CN, 20 CF 3 , -OCF 3 , -NO 2 , -OR 6 , -NR 56
R
57 and C,-C 6 -alkyl, Ra is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OC-C 6 -alkyl, -C(O)OCO-C6 alkyl, -C(=O)-R 62 , -COOH and -NH 2 , 25 R 6 1 is independently selected from halogen, -C(O)OC 1
-C
6 -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , NO 2 , -OH, -OCI-C 6 -alkyl, -NH 2 , C(=O) or Cl-C6-alkyl,
R
62 is C 1
-C
6 -alkyl, aryl optionally substituted with one or more substituents independently se lected from halogen, or heteroaryl optionally substituted with one or more Cl-C 6 -alkyl inde 30 pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. In another embodiment V is arylene, heteroarylene, or arylene-C 1
.C
6 -alkylene, wherein the 35 alkylene is optionally substituted with one or more substituents independently selected R5, WO 2004/080480 PCT/DK2004/000158 41 and the arylene or heteroarylene is optionally substituted with one or more substituents inde pendently selected from R". In another embodiment V is arylene, Heti, or arylene-Cl.C 6 -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R5, and the 5 arylene or heteroarylene moiety is optionally substituted with one or more substituents inde pendently selected from R". In another embodiment V is arylene, Het2, or arylene-Cl.C 6 -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde 10 pendently selected from R 55 . In another embodiment V is arylene, Het3, or arylene-C,.C 6 -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R5, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde pendently selected from R 55 . 15 In another embodiment V is arylene optionally substituted with one or more substituents in dependently selected from R 55 . In another embodiment V is ArG1 optionally substituted with one or more substituents inde pendently selected from R5 5 . In another embodiment V is phenylene, naphthylene or anthranylene optionally substituted 20 with one or more substituents independently selected from R 5 5 . In another embodiment V is phenylene optionally substituted with one or more substituents independently selected from R ss . In another embodiment R 5 is independently selected from halogen, CI-C 6 -alkyl, -CN, -OCF 3
,-CF
3 , -NO 2 , -OR" ,
-NR
56
R
57 , -NR 56 C(O)R 57 25 -SR 56 , -OC 1
-C
8 -alkyl-C(O)OR 5 6 , or -C(O)OR", * CI-C 6 -alkyl optionally substituted with one or more substituents independently se lected from
R
5 8 * aryl, aryl-Ci-C6-alkyl, heteroaryl, or heteroaryl-Cl-C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substitu 30 ents independently selected from R 5 . In another embodiment R 55 is independently selected from *halogen, C 1
-C
6 -alkyl, -CN, -OCF 3
,-CF
3 , -NO 2 , -OR 56 , -NR 6
R
57 , -NR 56
C(O)R
57
-SR
5 6 , -OC 1
-C
8 -alkyl-C(O)OR 5 ", or -C(O)OR 6 * C 1 -C6-alkyl optionally substituted with one or more substituents independently se 35 lected from R 58 WO 2004/080480 PCT/DK2004/000158 42 eArG1, ArG1-Cl-C 6 -alkyl, Het3, or Het3-Cl-Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu ents independently selected from R 59 . 5 In another embodiment R 55 is independently selected from halogen, -OR 6 , -NR 6
R
5 7 ,
-C(O)OR
5 6 8 , -OC 1
-C
8 -alkyl-C(O)ORm ,
-NR
56
C(O)R
57 or C-Cs-alkyl. In another embodiment R 55 is independently selected from halogen, -OR , -NR 56
R
57 , -C(O)OR", -OC,-Ca-alkyl-C(O)OR", -NR"C(O)R 7 , methyl or ethyl. In another embodiment R 5 ' and R s7 are independently selected from hydrogen, CF 3 , 10 Cl-C 12 -alkyl, or -C(=O)-Cj-C 6 -alkyl; R 56 and R 5 7 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. In another embodiment R' and R1 7 are independently selected from hydrogen or
CI-C
12 -alkyl, R 56 and R 5 7 when attached to the same nitrogen atom may form a 3 to 8 mem bered heterocyclic ring with the said nitrogen atom. 15 In another embodiment R6 and R 5 7 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 5 6 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. In another embodiment CGr is N H N N 20 wherein AA is C l
-C
6 -alkylene, arylene, heteroarylene, arylene-ClC 6 -alkylene or arylene-C 2 Ce-alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 63 , and the arylene or heteroarylene is optionally 25 substituted with one or more substituents independently selected from R 6 4 ,
R
63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
R
4 is independently selected from 30 *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 5 , -NR 65
R
66 , -SR 6 5
,
WO 2004/080480 PCT/DK2004/000158 43
-NR
65
S(O)
2
R
66 , -S(O) 2
NR
5 Rr 6 , -S(O)NR 5
R
6 6 , -S(O)R 65 , -S(O) 2
R
6 5 , -OS(0) 2
R
65 , -C(0)NR 6
R
6 , -OC(O)NR"R", -NR 65
C(O)R
66 , -CH 2
C(O)NR
65
R
66 , -OC-C 6 alkyl-C(O)NR 6 5 R 66 , -CH 2 0R 65 , -CH 2
OC(O)R
65 , -CH 2
NR
6 5
R
8 , -OC(O)R 65 , -OC1-C6 alkyl-C(O)OR 65 , -OC1-C 5 -alkyl-OR 6 5 , -SCj-C 6 -alkyl-C(O)OR 65 , -C 2
-C
6 -alkenyl 5 C(=O)OR 65 , -NR 65 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 5 , -NR 6 -C(=O)-Cl-C6 alkenyl-C(=O)OR , -C(O)OR 6 s , or-C 2
-C
6 -alkenyl-C(=O)R, * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, each of which may optionally be substi tuted with one or more substituents selected from R 67 , 10 * aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C l
-C
6 -alkoxy, aryl-C l
-C
6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl
C
6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl or heteroaryl-C 2
-C
6 -alkynyl, 15 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 68 ,
R
6 and R 66 are independently selected from hydrogen, OH, CF 3 , Cl-C 1 2 -alkyl, aryl-Cl-Ce alkyl, -C(=O)-R 6 9 , aryl or heteroaryl, wherein the alkyl groups may optionally be substituted 20 with one or more substituents selected from R 70 , and the aryl and heteroaryl groups may op tionally be substituted with one or more substituents independently selected from R 71 ; R 65 and R 66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or 25 two double bonds,
R
67 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 65 , and -NR 65
R
66 ,
R
6 8 is independently selected from halogen, -C(O)OR 5 , -CH 2 C(0)OR 6 s , -CH 2 0R 65 , -CN, 30 CF 3 , -OCF 3 , -NO 2 , -OR 6 5 , -NR 65
R
66 and Cl-C 6 -alkyl,
R
69 is independently selected from C 1
-C
6 -alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more CI-C 6 -alkyl, WO 2004/080480 PCT/DK2004/000158 44
R
70 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OCl 1
-C
6 -alkyl, -C(O)OC 1
-C
6 alkyl, -COOH and -NH 2 ,
R
71 is independently selected from halogen, -C(O)OCI-C6-alkyl, -COOH, -CN, -CF 3 , -OCF 3 , 5 NO 2 , -OH, -OC,-C 6 -alkyl, -NH 2 , C(=O) or C 1
-C
6 -alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. In another embodiment AA is arylene, heteroarylene or arylene-Cl.C 6 -alkylene, wherein the 10 alkylene is optionally substituted with one or more R 63 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 64 . In another embodiment AA is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 4 . In another embodiment AA is ArG1 or Hetl optionally substituted with one or more substitu 15 ents independently selected from R6. In another embodiment AA is ArG1 or Het2 optionally substituted with one or more substitu ents independently selected from R 64 . In another embodiment AA is ArG1 or Het3 optionally substituted with one or more substitu ents independently selected from R 64 20 In another embodiment AA is phenylene, naphtylene, anthrylene, carbazolylene, thienylene, pyridylene, or benzodioxylene optionally substituted with one or more substituents independ ently selected from R 64 In another embodiment AA is phenylene or naphtylene optionally substituted with one or more substituents independently selected from R 4 . 25 In another embodiment R 64 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 ,
-OR
65 , -NR 65
R
66 , C-C 6 -alkyl, -OC(O)R 6 5 , -OC 1 -Co-alkyl-C(O)OR 5 , aryl-C2-C 6 -alkenyl, aryloxy or aryl, wherein C-C 6 -alkyl is optionally substituted with one or more substituents independ ently selected from R 67 , and the cyclic moieties optionally are substituted with one or more substituents independently selected from R 68 . 30 In another embodiment R 64 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 5 ,
-NR
65
R
66 , methyl, ethyl, propyl, -OC(O)R 6 5 , -OCH 2
-C(O)OR
65 , -OCH 2
-CH
2
-C(O)OR
65 , phenoxy optionally substituted with one or more substituents independently selected from
R
6 8
.
WO 2004/080480 PCT/DK2004/000158 45 In another embodiment R 65 and R 6 " are independently selected from hydrogen, CFa, Cl-C1 2 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents inde pendently selected from R 71 . In another embodiment R" 5 and R 66 are independently hydrogen, Cl-C, 2 -alkyl, aryl, or het 5 eroaryl optionally substituted with one or more substituents independently selected from R 7 . In another embodiment R" 5 and R" are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Hetl optionally substituted with one or more substituents inde pendently selected from R 71 In another embodiment R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 10 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents inde pendently selected from R 7 . In another embodiment R 6 5 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1. or Het3 optionally substituted with one or more substituents inde pendently selected from R". 15 In another embodiment R 6 s and R 6 6 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R 71 independently; or isoxazolyl optionally substituted with one or more substituents independ ently selected from R 71 In another embodiment R 71 is halogen or Cl-C 6 -alkyl. 20 In another embodiment R 7 1 is halogen or methyl. In another embodiment Frgl consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. In another embodiment Frgl consists of 0 to 5 Gly. In another embodiment Frgl consists of 0 Gly. 25 In another embodiment Frgl consists of 1 Gly. In another embodiment Frgl consists of 2 Gly. In another embodiment Frgl consists of 3 Gly. In another embodiment Frgl consists of 4 Gly. In another embodiment Frgl consists of 5 Gly. 30 In another embodiment GB is of the formula B'-B 2 -C(O)-, B 1
-B
2
-SO
2 - or B'-B 2
-CH
2 -, wherein B' and B 2 are as defined in claim 1. In another embodiment GB is of the formula B'-B 2 -C(O)-, B'-B 2
-SO
2 - or B 1
-B
2 -NH-, wherein B' and B 2 are as defined in claim 1. In another embodiment GB is of the formula B'-B 2 -C(O)-, B 1
-B
2
-CH
2 - or B'-B 2 -NH-, wherein 35 B' and B 2 are as defined in claim 1.
WO 2004/080480 PCT/DK2004/000158 46 In another embodiment GB is of the formula B'-B 2
-CH
2 -, B'-B 2 -SO2- or B'-B 2 -NH-, wherein B' and B 2 are as defined in claim 1. In another embodiment GB is of the formula B'-B 2 -C(0)- or B'-B 2 -SO2-, wherein B' and B 2 are as defined in claim 1. 5 In another embodiment GB is of the formula B'-B 2 -C(O)- or B'-B 2
-CH
2 -, wherein B' and B 2 are as defined in claim 1. In another embodiment GB is of the formula B'-B 2 -C(0)- or B 1
-B
2 -NH-, wherein B' and B 2 are as defined in claim 1. In another embodiment G3 is of the formula B'-B 2
-CH
2 - or B'-B 2
-SO
2 -, wherein B' and B 2 are 10 as defined in claim 1. In another embodiment G B is of the formula B'-B 2 -NH- or B'-B 2
-SO
2 -, wherein B 1 and B 2 are as defined in claim 1. In another embodiment G B is of the formula B'-B 2
-CH
2 - or B'-B 2 -NH-, wherein B' and B 2 are as defined in claim 1. 15 In another embodiment GB is of the formula B'-B 2 -C(0)-. In another embodiment GB is of the formula B'-B 2
-CH
2 -. In another embodiment G B is of the formula B'-B 2 -SO2-. In another embodiment GB is of the formula B'-B 2 -NH-. In another embodiment B' is a valence bond, -0-, or -S-. 20 In another embodiment B' is a valence bond, -0-, or -N(R)-. In another embodiment B' is a valence bond, -S-, or -N(R6)-. In another embodiment B' is -0-, -S- or -N(Rr)-. In another embodiment B' is a valence bond or -0-. In another embodiment B' is a valence bond or -S-. 25 In another embodiment B' is a valence bond or -N(R 6 )-. In another embodiment B' is -O-or -S-. In another embodiment B' is -O-or -N(R6)-. In another embodiment B' is -S-or -N(R 6 )-. In another embodiment B' is a valence bond. 30 In another embodiment B' is -0-. In another embodiment B' is -S-. In another embodiment B' is -N(R 6 )-. In another embodiment B 2 is a valence bond, Cl-C, 8 -alkylene, C 2 -C1 8 -alkenylene, C 2 -C1 8 alkynylene, arylene, heteroarylene, -C,-C1 8 -alkyl-aryl-, -C(=0)-Cl-Cs-alkyl-C(=0)-, -C(=0) 35 Cl-C, 8 -alkyl-O-Cl-Cls-alkyl-C(=0)-, -C(=O)-Cl-C 1 is-alkyl-S-Cl-Cl, 8 -alkyl-C(=0)-, -C(=0)-Cl- WO 2004/080480 PCT/DK2004/000158 47 Cia-alkyl-NRP 6 -Cl-Cs-alkyl-C(=O)-; and the alkylene and arylene moieties are optionally sub stituted as defined in claim 1. In another embodiment B 2 is a valence bond, C 1
-C
8 is-alkylene, C 2
-C
1 8 -alkenylene, C 2
-C
18 alkynylene, arylene, heteroarylene, -Cl-C 1 8 -alkyl-aryl-, -C(=O)-C 1
-C
1 8 -alkyl-C(=O)-, -C(=O) 5 Cl-Cs-alkyl-O-C l
-C,
8 -alkyl-C(=O)-, and the alkylene and arylene moieties are optionally sub stituted as defined in claim 1. In another embodiment B 2 is a valence bond, Cl-C, 8 -alkylene, C 2 -CI-alkenylene, C 2
-C
1 5 8 alkynylene, arylene, heteroarylene, -C-C, 8 -alkyl-aryl-, -C(=O)-C 1
-C
18 -alkyl-C(=O0)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. 10 In another embodiment B 2 is a valence bond, Cl-C, 8 -alkylene, arylene, heteroarylene, -Cl
C
18 -alkyl-aryl-, -C(=O)-C 1
-C
18 -alkyl-C(=O)-, and the alkylene and arylene moieties are option ally substituted as defined in claim 1. In another embodiment B 2 is a valence bond, Cl-Cls-alkylene, arylene, heteroarylene, -C,
C
18 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in 15 claim 1. In another embodiment B 2 is a valence bond, Cl-C,-alkylene, arylene, -Cl-Ci 8 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment B 2 is a valence bond or -Cl-C 1 8 -alkylene, and the alkylene moieties are optionally substituted as defined in claim 1. 20 In another embodiment Frg2 comprises 1 to 16 positively charged groups. In another embodiment Frg2 comprises 1 to 12 positively charged groups. In another embodiment Frg2 comprises 1 to 10 positively charged groups. In another embodiment Frg2 is a fragment containing basic amino acids independently se lected from the group consisting of Lys and Arg and D-isomers of these. 25 In another embodiment the basic amino acid is Arg. In another embodiment X is -OH or -NH 2 . In another embodiment X is -NH 2 . In another embodiment the pharmaceutical preparation further comprises at least 3 phenolic molecules. 30 In another embodiment the acid-stabilised insulin is selected from the group consisting of A21G A21G, B28K, B29P A21G, B28D A21G, B28E 35 A21G, B3K, B29E WO 2004/080480 PCT/DK2004/000158 48 A21G, desB27 A21G, B9E A21G, B9D A21G, B10E 5 In another embodiment zinc ions are present in an amount corresponding to 10 to 40 ag Zn/100 U insulin In another embodiment zinc ions are present in an amount corresponding to 10 to 26 4g Zn/100 U insulin. In another embodiment the ratio between insulin and the zinc-binding ligand of the invention 10 is in the range from 99:1 to 1:99. In another embodiment the ratio between insulin and the zinc-binding ligand of the invention is in the range from 95:5 to 5:95 In another embodiment the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 80:20 to 20:80 15 In another embodiment the ratio between between insulin and the zinc-binding ligand of the invention is in the range from 70:30 to 30:70 In another aspect the invention relates to a method of preparing a zinc-binding ligand of the invention comprising the steps of * Identifying starter compounds that binds to the R-state HisBliO-Zn 2 + site 20 * optionally attaching a fragment consisting of 0 to 5 neutral a- or 3-amino acids Attaching a fragment comprising 1 to 20 positively charged groups independently se lected from amino or guanidino groups In another aspect the invention relates to a method of prolonging the action of an acid stabilised insulin preparation which comprises adding a zinc-binding ligand of the invention to 25 the acid-stabilised insulin preparation. In another aspect the invention relates to a method of treating type 1 or type 2 diabetes com prising administering to a patient in need thereof a theraputically effective amount of a phar maceutical preparation comprising 1. Acid-stabilised insulin 30 2. Zinc ions 3. A zinc-binding ligand that binds to the R-state HisB 1 o-Zn2+ site, where said ligand may be as described in the embodiments above. In another aspect the invention provides an embodiment 1,which is a pharmaceutical prepa 35 ration comprising WO 2004/080480 PCT/DK2004/000158 49 1. Acid-stabilised insulin 2. Zinc ions 3. A zinc-binding ligand of the following general formula (I) 5 CGr-Lnk-Frgl-Frg2-X (I) wherein: CGr is a chemical group which reversibly binds to a His B 0 Zn2+ site of an insulin hexamer; 10 Lnk is a linker selected from *a valence bond Sa chemical group Go of the formula -B 1
-B
2 -C(O)-, -B 1
-B
2 -SO2-, -B 1
-B
2
-CH
2 -, or -B 1 B 2 -NH-; wherein B1 is a valence bond, -0-, -S-, or -NR 6 -, 15 B2 is a valence bond, Cl-Ca 18 -alkylene, C 2
-C
18 -alkenylene, C 2
-C
1 8 -alkynylene, arylene, heteroarylene, -Cl-C, 18 -alkyl-aryl-, -C 2
-C
1 8 -alkenyl-aryl-, -C2-C1 8 -alkynyl-aryl-, -C(=O) Cj-C 1 8 -alkyl-C(=O)-, -C(=O)-Cj-C 18 -alkenyl-C(=O)-, -C(=O)-Cj-C 18 -alkyl-O-Cj-Cs 8 alkyl-C(=0)-, -C(=O)- Cl-C 1 8 -alkyl-S-C 1
-C
18 -alkyl-C(=O)-, -C(=O)-Cj-C, 8 -alkyl-NR 6 -Cj Cja-alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C(=O)-; 20 wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF 3 , -OR 6B , or -NR 6
BR
7 B and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR 6 B , -C(O)H, OCOR 6B , -SO 2 , -CN, -CF 3 ,
-OCF
3 , -NO 2 , -OR 6 B , -NR"BR 7 B,
C
1 -C18-alkyl, or Cl-C 1 a-alkanoyl;
R
6 ' and R 7 B are independently H, C-C 4 -alkyl; 25 FrgI is a fragment consisting of 0 to 5 neutral a- or p-amino acids Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups; and 30 X is -OH, -NH 2 or a diamino group, or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric forms.
WO 2004/080480 PCT/DK2004/000158 50 Embodiment 2. A pharmaceutical preparation according to embodiment 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano 1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5 5 mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbitu rates, naphthoic acids and salicylic acids. Embodiment 3. A pharmaceutical preparation according to embodiment 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2 napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4 10 cyano-1,2,3-triazoles. Embodiment 4. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is X H HN / 0 N O A / H or HN N-B or
R
3 A O/ i 4 N R HN R3A 0 2 0 R R IA 0 R2A wherein 15 Xis=0,=Sor=NH Y is -S-, -0- or -NH
R
1 , R 1 A and R 4 are independently selected from hydrogen or C 1
-C
6 -alkyl,
R
2 and R 2A are hydrogen or Cl-C 6 -alkyl or aryl, R 1 and R 2 may optionally be combined to 20 form a double bond, R1A and R2A may optionally be combined to form a double bond,
R
3 , R 3 A and R 5 are independently selected from hydrogen, halogen, aryl optionally substi tuted with one or more substituents independently selected from Rl 6 , CI-C 6 -alkyl, or -C(0)NR"R 12 , 25 A, A' and B are independently selected from Cl-C 6 -alkyl, aryl, aryl-Cl-C 6 -alkyl, -NR"-aryl, aryl-C 2
-C
6 -alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 6 and the aryl or heteroaryl is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 , A and R 3 may be connected through one or two valence bonds, B and R 5 may be connected 30 through one or two valence bonds,
R
6 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
,
WO 2004/080480 PCT/DK2004/000158 51
R
7 , R 8 , R 9 and R 10 are independently selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 11 , -NR 11
R
12 , -SR 11 , 5 -NR"S(O) 2
R
12 , -S(O) 2
NR"
1
R
12 , -S(O)NR 11
R
1 2 , -S(O)R", -S(O) 2 R", -OS(0) 2 RI", -C(0)NR"R 12 , -OC(O)NR"R 12 , -NR"C(O)R l , -CH 2
C(O)NR
1 1 2R,
-OC
1
-C
6 -alkyl-C(O)NR 1 1
R
2 , -CH 2 0R 11 , -CH20C(O)R 1 , -CH 2
NR
11
"R
1 2 , -OC(O)R 11 ,
-OC
1
-C
1 5 -alkyl-C(O)OR 1 1 , -OCl-C 6 -alkyl-OR 1 , -SC 1
-C
6 -alkyl-C(O)OR 11 "
-C
2
-C
6 -alkenyl-C(=O)OR", -N R 1 "-C(=O)-Cl-C 6 -alkyl-C(=O)OR 1 , 10 -NR 1
"-C(=O)-C
1
-C
6 -alkenyl-C(=O)OR 1 1 , -C(0)OR" 1 , C(O)R" 1 , or -C 2
-C
6 -alkenyl
C(=O)R
1 , =0, or -C 2
-C
6 -alkenyl-C(=O)-NR"R 1 2 , * C 1 -Ce-alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, each of which may optionally be substi tuted with one or more substituents independently selected from R 13 , 15 earyl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl
C
6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl, heteroaryl-C 2
-C
6 -alkynyl, or C 3
-C
6 cycloalkyl, 20 of which each cyclic moiety may optionally be substituted with one or more substitu ents independently selected from R",
R
1 1 and R 12 are independently selected from hydrogen, OH, CI-C 20 -alkyl, aryl-Cl-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents 25 independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 1 6 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 30
R
13 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 11 , -C(0)OR 1 , -NR"R 12, and -C(O)NR 11
R
12 ,
R
14 is independently selected from halogen, -C(O)OR 11 , -CH 2
C(O)OR
11 , -CH 2 0R 11 , -CN, 35 CF3, -OCF 3 , -NO 2 , -OR 11 , -NR 11
R
1 2 , -NR"C(O)R 1 , -S(O) 2
R
11 , aryl and C,-C 6 -alkyl, WO 2004/080480 PCT/DK2004/000158 52
R
15 is independently selected from halogen, -CN, -CF3, =0, -OCFa, -OC,-C 6 -alkyl, -C(0)OC,
C
6 -alkyl, -COOH and -NH 2 , 5 R 16 is independently selected from halogen, -C(O)OCI-C 6 -alkyl, -COOH, -CN, -CF3, -OCF 3 , NO 2 , -OH, -OCI-C 6 -alkyl, -NH 2 , C(=O) or Cl-Ce-alkyl, or any enantiomer, diastereomer, in cluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically accept able acid or base. Embodiment 5. A pharmaceutical composition according to embodiment 4 wherein X is =0 or 10 =S. Embodiment 6. A pharmaceutical composition according to embodiment 5 wherein X is =0. Embodiment 7. A pharmaceutical composition according to embodiment 5 wherein X is =S. Embodiment 8. A pharmaceutical composition according to any one of the embodiment s 4 to 7 wherein Y is -0- or -S-. 15 Embodiment 9. A pharmaceutical composition according to embodiment 8 wherein Y is -0-. Embodiment 10. A pharmaceutical composition according to embodiment 8 wherein Y is -NH-. Embodiment 11. A pharmaceutical composition according to embodiment 8 wherein Y is -S-. Embodiment 12. A pharmaceutical composition according to any one of the embodiment s 4 20 to 11 wherein A is aryl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or different. Embodiment 13. A pharmaceutical composition according to embodiment 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R'o which may be the same or different. 25 Embodiment 14. A pharmaceutical composition according to embodiment 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R 7 , R 8 , R , and Ro which may be the same or different. Embodiment 15. A pharmaceutical composition according to embodiment 14 wherein A is R9 R9 or
R
7 R S Rm 30 16. A pharmaceutical composition according to embodiment 14 wherein A is phenyl.
WO 2004/080480 PCT/DK2004/000158 53 Embodiment 17. A pharmaceutical composition according to any one of the embodimernt s 4 to 11 wherein A is heteroaryl optionally substituted with up to four substituents, R 7 , R", R 9 , and R 10 which may be the same or different. Embodiment 18. A pharmaceutical composition according to embodiment 17 wherein A is 5 selected from Hetl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or different. Embodiment 19. A pharmaceutical composition according to embodiment 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or different. 10 Embodiment 20. A pharmaceutical composition according to embodiment 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R 7 , R 8 , R", and R 10 which may be the same or different. Embodiment 21. A pharmaceutical composition according to embodiment 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl, 15 wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R9, and Ro which may be the same or different. Embodiment 22. A pharmaceutical composition according to embodiment 20 wherein A is benzofuranyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different. 20 Embodiment 23. A pharmaceutical composition according to embodiment 22 wherein A is R R 7 or R 8 or o R 7 24. A pharmaceutical composition according to embodiment 20 wherein A is carbazolyl op tionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different. 25 Embodiment 25. A pharmaceutical composition according to embodiment 24 wherein A is
R
9 RH N
R
7 WO 2004/080480 PCT/DK2004/000158 54 Embodiment 26. A pharmaceutical composition according to embodiment 20 wherein A is quinolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 o which may be the same or different. Embodiment 27. A pharmaceutical composition according to embodiment 26 wherein A is
R
9
R
9 N I or N 5
R
7
R
8 R Embodiment 28. A pharmaceutical composition according to embodiment 20 wherein A is indolyl optionally substituted with up to four substituents R 7 , R 8 , Rg, and R 1 o which may be the same or different. Embodiment 29. A pharmaceutical composition according to embodiment 28 wherein A is R R R8
R
9 R 8 NH Or NH 10 R7 R Embodiment 30. A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R' is hydrogen. Embodiment 31. A pharmaceutical composition according to any one of the embodiment s 4 to 30 wherein R 2 is hydrogen. 15 Embodiment 32. A pharmaceutical composition according to any one of the embodiment s 4 to 29 wherein R 1 and R 2 are combined to form a double bond. Embodiment 33. A pharmaceutical composition according to any one of the embodiment s 4 to 32 wherein R 3 is C 1
-C
6 -alkyl, halogen, or C(O)NR"R 7 . Embodiment 34. A pharmaceutical composition according to embodiment 33 wherein R 3 is 20 Cl-C 6 -alkyl or C(O)NR"R 17 . Embodiment 35. A pharmaceutical composition according to embodiment 34 wherein R 3 is methyl. Embodiment 36. A pharmaceutical composition according to any one of the embodiment s 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and 25 R 10 which may be the same or different. Embodiment 37. A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 wherein R 4 is hydrogen.
WO 2004/080480 PCT/DK2004/000158 55 Embodiment 38. A pharmaceutical composition according to any one of the embodiment s 4 to 11 or 36 to 37 wherein R 5 is hydrogen. Embodiment 39. A pharmaceutical composition according to any one of the embodiment s 4 to 38 wherein R 6 is aryl. 5 Embodiment 40. A pharmaceutical composition according to embodiment 39 wherein R 6 is phenyl. Embodiment 41. A pharmaceutical composition according to any one of the embodiment s 4 to 40 wherein R 7 , R 8 , R 9 and R 10 are independently selected from 10 hydrogen, halogen, -NO 2 , -OR 1 1 , -NR"R' 2 , -SR 11 , -NR 11
S(O)
2
R
12 , -S(O) 2
NR"R
12
-S(O)NR"R
12 , -S(O)R 11 , -S(O) 2
R
11 , -OS(O) 2
R
11 , -NR"C(O)R 12 , -CH 2 0R 1 1 , CH 2
OC(O)R
1 , -CH 2
NR"R
12 , -OC(O)R 11 , -OC,-C 6 -alkyl-C(O)OR 1 , -OC-Co alkyl-C(O)NR"R 12 , -OC,-C 6 -alkyl-OR 1 , -SC 1
-C
6 -alkyl-C(O)OR 1 , -C 2
-C
6 -alkenyl
C(=O)OR
1 , -C(O)OR 1 1 , or -C 2
-C
6 -alkenyl-C(=O)R 11 , 15 * Cl-C 6 -alkyl, C 2 -C6-alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted with one or more substituents independently selected from R 13 *aryl, aryloxy, aroyl, arylsulfanyl, aryl-Cl-C6-alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 20 C 6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroary-Cl-C 6 -alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 . Embodiment 42. A pharmaceutical composition according to embodiment 41 wherein R 7 , R 8 ,
R
9 and R 1 0 are independently selected from 25 hydrogen, halogen, -NO 2 , -OR 11 , -NR 11 R12, -SR 11 , -S(O) 2
R
11 , -OS(O) 2
R
11 , CH 2 OC(O)R", -OC(O)R", -OCI-C 6 -alkyl-C(O)OR 1 , -OCj-C 6 -alkyl-OR 1 , -SC 1
-C
6 alkyl-C(O)OR", -C(O)OR", or -C 2
-C
6 -alkenyl-C(=O)R 1 1 , 30 * C-C 6 -alkyl or Cl-C 6 -alkenyl which may each optionally be substituted with one or more substituents independently selected from R 13 * aryl, aryloxy, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-C l
-C
6 -alkyl, heteroaryl, WO 2004/080480 PCT/DK2004/000158 56 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 Embodiment 43. A pharmaceutical composition according to embodiment 42 wherein R', R 8 ,
R
9 and R 10 are independently selected from 5 *hydrogen, halogen, -NO 2 , -OR 11 , -NR"R 12 , -SR", -S(O) 2
R"
1 , -OS(O) 2
R"
1 , CH 2
OC(O)R
11 , -OC(O)R" 1 , -OCI-C 6 -alkyl-C(O)OR 1 , -OC 1
-C
6 -alkyl-OR 11 , -SC 1
-C
6 alkyl-C(O)OR 11 , -C(O)OR 1 , or -C 2
-C
6 -alkenyl-C(=O)R 1 , 10 *Cl-C 6 -alkyl or Cl-C 6 - which may each optionally be substituted with one or more substituents independently selected from R" 3 *aryl, aryloxy, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, 15 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 Embodiment 44. A pharmaceutical composition according to embodiment 43 wherein R 7 , R 8 ,
R
9 and R 1 0 are independently selected from *hydrogen, halogen, -OR", -OCI-C 6 -alkyl-C(O)OR" 1 , or -C(0)OR 1 , 20 * C-C6-alkyl which may each optionally be substituted with one or more substituents independently selected from R 13 *aryl, aryloxy, aryl-Cl-C 6 -alkoxy, 25 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 4 . Embodiment 45. A pharmaceutical composition according to embodiment 44 wherein R 7 , R 8 ,
R
9 and R 10 are independently selected from 30 * hydrogen, halogen, -OR 1 ", -OC 1
-C
6 -alkyl-C(O)OR 1 , or -C(O)OR 11 , * C 1
-C
6 -alkyl which may each optionally be substituted with one or more substituents independently selected from R 13 35 eArG1, ArGloxy, ArG1-C 1
-C
6 -alkoxy, WO 2004/080480 PCT/DK2004/000158 57 of which each of the cyclic moieties optionally may be substituted with one or more substitu ents independently selected from R 1 4 Embodiment 46. A pharmaceutical composition according to embodiment 45 wherein R 7 , R 8 , 5 R 9 and R 1 o are independently selected from *hydrogen, halogen, -OR 11 , -OC-C 6 -alkyl-C(O)OR" 1 , or -C(O)OR 1 , * Cl-C 6 -alkyl which may optionally be substituted with one or more substituents inde pendently selected from R1 3 10 * phenyl, phenyloxy, phenyl-C-C 6 -alkoxy, wherein each of the cyclic moieties option ally may be substituted with one or more substituents independently selected from
R
1 4 . Embodiment 47. A pharmaceutical composition according to any one of the embodiment s 4 15 to 46 wherein R 1 and R 1 2 are independently selected from hydrogen, C 1
-C
2 0 -alkyl, aryl or aryl-C 1
-C
6 -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substi tuted one or more substituents independently selected from R' 6 ; R" and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro 20 gen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds. Embodiment 48. A pharmaceutical composition according to embodiment 47 wherein R 1 1 and
R
12 are independently selected from hydrogen, Cl-C 2 0 -alkyl, aryl or aryl-Cl-Co-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently 25 selected from R 15 , and the aryl groups may optionally be substituted one or more substitu ents independently selected from R 16 . Embodiment 49. A pharmaceutical composition according to embodiment 48 wherein R 1 1 and
R
2 are independently selected from phenyl or phenyl-Cl-Ce-alkyl. Embodiment 50. A pharmaceutical composition according to embodiment 48 wherein one or 30 both of R 1 and R 1 2 are methyl. Embodiment 51. A pharmaceutical composition according to any one of the embodiment s 4 to 50 wherein R 13 is independently selected from halogen, CF 3 , OR" 1 or NR"R 2 . Embodiment 52. A pharmaceutical composition according to embodiment 51 wherein R1 3 is independently selected from halogen or OR".
WO 2004/080480 PCT/DK2004/000158 58 Embodiment 53. A pharmaceutical composition according to embodiment 52 wherein R 13 is
OR
11 . Embodiment 54. A pharmaceutical composition according to any one of the embodiment s 4 to 53 wherein R' 4 is independently selected from halogen, -C(O)OR 11 , -CN, -CF 3 , -OR" 1 , 5 S(O) 2
R
11 , and C 1
-C
8 -alkyl. Embodiment 55. A pharmaceutical composition according to embodiment 54 wherein R 14 is independently selected from halogen, -C(O)OR" 1 , or -OR". Embodiment 56. A pharmaceutical composition according to any one of the embodiment s 4 to 55 wherein R 15 is independently selected from halogen, -CN, -CF3, -C(O)OC 1
-C
6 -alkyl,and 10 -COOH. Embodiment 57. A pharmaceutical composition according to embodiment 56 wherein R 15 is independently selected from halogen or -C(O)OC,-C 6 -alkyl. Embodiment 58. A pharmaceutical composition according to any one of the embodiment s 4 to 57 wherein R 16 is independently selected from halogen, -C(O)OCl-C 6 -alkyl, -COOH, -NO 2 , 15 -OC-C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl. Embodiment 59. A pharmaceutical composition according to embodiment 58 wherein R 1 6 is independently selected from halogen, -C(O)OC,-C 6 -alkyl, -COOH, -NO 2 , or Cl-C 6 -alkyl. Embodiment 60. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is 17 N//N N D NR- 2 6 F or N E or NF" G N, 18 NN H R H 19 H O R/ H6 20 N-, . I 2 or N N D1-G' 20 H wherein
R
19 is hydrogen or C 1
-C
6 -alkyl,
R
20 is hydrogen or C 1 -Co-alkyl, 25 D, D' and F are a valence bond, Cl-C 6 -alkylene or C-C-alkenylene optionally substituted with one or more substituents independently selected from R 7 2 ,
R
72 is independently selected from hydroxy, CI-C6-alkyl, or aryl, WO 2004/080480 PCT/DK2004/000158 59 E is Cl-CB-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R 21 , R 2 2 and R 23 G and G 1 are C 1
-C
6 -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub 5 stituted with up to three substituents R 24 , R 25 and R 26 ,
R
17 , R'8, R 2 1 , R 22 , R 23 , R 24 , R 25 and R 26 are independently selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , 10 -OCF 2
CHF
2 , -S(O) 2
CF
3 , -SCF 3 , -NO 2 , =0, -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
S(O)
2
R
28 ,
-S(O)
2
NR
2 7
R
28 , -S(O)NR 27
R
28 , -S(O)R 2 7 , -S(O) 2
R
2 7 , -C(O)NR 27
R
28 , -OC(O)NR 27
R
28 ,
-NR
27
C(O)R
28 , -NR 27
C(O)OR
28 , -CH 2
C(O)NR
27
R
28 , -OCH 2
C(O)NR
27
R
2 8 , -CH 2 0R 27 ,
-CH
2
NR
27
R
28 , -OC(O)R 27 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC 1
-C
6 -alkyl-C(O)OR 27 , -C 2
-C
6 alkenyl-C(=O)OR 2 7 , -NR 27 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 27 , -NR 27
-C(=O)-C
1
-C
6
_
15 alkenyl-C(=O)OR 27 , -C(=O)NR 27 -C -C 6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 27 ,0or
-C(O)OR
27 * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, 20 which may optionally be substituted with one or more substituents independently se lected from R 29 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C6-alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2
-C
6 25 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 , 30
R
27 and R 28 are independently selected from hydrogen, C 1
-C
6 -alkyl, aryl-C 1
-C
6 -alkyl or aryl, or
R
27 and R 28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two 35 double bonds, WO 2004/080480 PCT/DK2004/000158 60
R
2 9 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 27 , and -NR 27
R
28
R
3 o is independently selected from halogen, -C(O)OR 27 , -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 27 , 5 -NR 27
R
2 8 and Cl-C 6 -alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 61. A pharmaceutical composition according to embodiment 60 wherein D is a valence bond. Embodiment 62. A pharmaceutical composition according to embodiment 60 wherein D is 10 Cl-C 6 -alkylene optionally substituted with one or more hydroxy, C 1
-C
6 -alkyl, or aryl. Embodiment 63. A pharmaceutical composition according to any one of the embodiment s 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 . Embodiment 64. A pharmaceutical composition according to embodiment 63 wherein E is 15 aryl optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 . Embodiment 65. A pharmaceutical composition according to embodiment 64 wherein E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 20 Embodiment 66. A pharmaceutical composition according to embodiment 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R 21 ,
R
22 and R 23 Embodiment 67. A pharmaceutical composition according to embodiment 66 wherein CGr is R 2 2 N 21 N R N R H 25 Embodiment 68. A pharmaceutical composition according to any one of the embodiment s 60 to 67 wherein R 2 1 , R2 and R 23 are independently selected from *hydrogen, halogen, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCHzCF 3 , -OCF 2
CHF
2 , -SCF 3 , NO 2 , -OR 27 , -NR 27
R
28 , -SR 27,
-C(O)NR
27
R
28 , -OC(O)NR 27 R", -NR 27
C(O)R
2 , 30 -NR 27
C(O)OR
28 , -CH 2
C(O)NR
27
R
28 , -OCH 2
C(O)NR
27
R
28 , -CH 2 0R 27 , -CH 2
NR
27
R
28 ,
-OC(O)R
27 , -OCl-C 6 -alkyl-C(O)OR 27 , -SCI-C 6 -alkyl-C(O)OR 2 7 , -C 2
-C
6 -alkenyl- WO 2004/080480 PCT/DK2004/000158 61
C(=O)
O R 2 7 , -NR27-C(=O)-Cs-C6-alkyl-C(=O)OR 27 ,
-NR
2 7
-C(=O)-C
1
-C
6 alkenyl-C(=O)OR 7 -, C(=O)NR27-C-6-alkyl-C(=O)OR27,-C 1
-C
6 -alkyl-C(=O)OR 2 7 , or -C(O)0R", 5 * C 1
-C
6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 29 10 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-Cs-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu 15 ents selected from R 30 . Embodiment 69. A pharmaceutical composition according to embodiment 68 wherein R 2 ,
R
22 and R 23 are independently selected from * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
2 8 , -SR 27 , -NR 27
C(O)R
28 , -NR 27 C(O)OR", 20 -OC(O)R 27 , -OCl-C6-alkyl-C(O)
O
R
27 , -SCI-C 6 -alkyl-C(O)OR 2 7 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 27 Cl -C 6 -alkyl-C(=O)OR 27 , -Cj-C 6 -alkyl-C(=O)OR 2 7 , or
-C(O)OR
27 * C 1
-C
6 -alkyl optionally substituted with one or more substituents independently se 25 lected from R 2 9 * aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-Ce alkyl, 30 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . Embodiment 70. A pharmaceutical composition according to embodiment 69 wherein R 21
R
22 and R 23 are independently selected from WO 2004/080480 PCT/DK2004/000158 62 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 2 7
R
2 8 , -SR 27 , -NR 2 7C(O)R28, -NR 27 C(0)OR 2 ,
-OC(O)R
27 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 27 -Cl-C-alkyl-C(=O)OR 27 , -C 1 -C6-alkyl-C(=O)OR 27 , or
-C(O)OR
7 , 5 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 2 9 * aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroary-Cl-C 6 10 alkyl of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R". Embodiment 71. A pharmaceutical composition according to embodiment 70 wherein R 21
R
2 and R 23 are independently selected from 15 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27 C(O)R8, -NR 27 C(0)OR 28 ,
-OC(O)R
27 , -OCI-C 6 -alkyl-C(O)OR 27 , -SC-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=0)NR 27
-C
1
-C
6 -alkyl-C(=0)OR 2 7 , -C 1 -Ce-alkyl-C(=0)OR 27 , or
-C(O)OR
2 7 20 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 * ArG1, ArG1-O-, ArG1-C(O)-, ArGl1-Cl-C 6 -alkoxy, ArG1-C-C 6 o-alkyl, Het3, Het3-Cj 25 C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . Embodiment 72. A pharmaceutical composition according to embodiment 71 wherein R 21 R2 and R 23 are independently selected from 30 * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
2 8 , -SR 27 , -NR 27
C(O)R
28 , -NR 27
C(O)OR
28
-OC(O)R
2 7 , -OCI-Co-alkyl-C(0)OR 27 , -SCI-C 6 -alkyl-C(0)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)N R 2 -C-C-alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=0)OR 7 , or
-C(O)OR
27 , 35 WO 2004/080480 PCT/DK2004/000158 63 * C-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 29 * phenyl, phenyloxy, phenyl-Cl-Cr-alkoxy, phenyl-Cl-C 6 -alkyl, 5 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30 . Embodiment 73. A pharmaceutical composition according to any one of the embodiment s 60 to 72 wherein R 19 is hydrogen or methyl. Embodiment 74. A pharmaceutical composition according to embodiment 73 wherein R9 is 10 hydrogen. Embodiment 75. A pharmaceutical composition according to any one of the embodiment s 60 to 74 wherein R 27 is Hydrogen, C 1
-C
6 -alkyl or aryl. Embodiment 76. A pharmaceutical composition according to embodiment 75 wherein R 27 is hydrogen or CI-C 6 -alkyl. 15 Embodiment 77. A pharmaceutical composition according to any one of the embodiment s 60 to 76 wherein R 28 is hydrogen or Cl-C 6 -alkyl. Embodiment 78. A pharmaceutical composition according to embodiment 60 wherein F is a valence bond. Embodiment 79. A pharmaceutical composition according to embodiment 60 wherein F is Cj 20 C-alkylene optionally substituted with one or more hydroxy, Cl-C 6 -alkyl, or aryl. Embodiment 80. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 79 wherein G is C 1
-C
6 -alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 . Embodiment 81. A pharmaceutical composition according to any one of the embodiment s 60 25 or 78 to 79 wherein G is Cl-C 6 -alkyl or ArG1, wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 . Embodiment 82. A pharmaceutical composition according to embodiment 80 wherein G is Cl-C6-alkyl. Embodiment 83. A pharmaceutical composition according to embodiment 82 wherein G is 30 phenyl optionally substituted with up to three substituents R 24 , R 2s and R 26 . Embodiment 84. A pharmaceutical composition according to any one of the embodiment s 60 to 83 wherein R 2 4 , R 25 and R 26 are independently selected from * hydrogen, halogen, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -SCF 3 , 35 NO 2 , -OR 27 , -NR 27
R
2 , -SR 27 , -C(O)NRR 28 , -OC(O)NRR 2 8 , -NR 2 7
C(O)R
8
,
WO 2004/080480 PCT/DK2004/000158 64
-NR
27
C(O)OR
28 , -CH 2
C(O)NR
27
R
28 , -OCH 2
C(O)NR
2 7
R
2 a , -CH 2 0R 27 , -CH 2 NR 2 7
R
2 8 ,
-OC(O)R
27 , -OCI-C 6 -alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -NR 27 -C(= O)-C-C 6 -alkyl-C(=O)OR 7 , -NRT-C(=O)-C -C 6 alkenyl-C(=O)OR 27 -, -C(=O)NR 27 -C 1
-C
6 -alkyl-C(=O)OR 27 , -Cl-Co-alkyl-C(=O)OR 27 , or 5 -C(O)OR, * CI-C6-alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se 10 lected from R 29 * aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, 15 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 Embodiment 85. A pharmaceutical composition according to embodiment 84 wherein R 24 ,
R
2 5 and R 2 6 are independently selected from 20 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
2 8 , -NR 27
C(O)OR
8 ,
-OC(O)R
27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC-C 6 -alkyl-C(O)OR 2 7 , -C 2 -C 6-alkenyl
C(=O)OR
2 7 , -C(=O)NR 27 -Cl-C 6 -alkyl-C(=O)OR 2 7 , -C 1
-C
6 -alkyl-C(=O)OR 2 7 , or
-C(O)OR
27 25 * CI-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 2 30 * aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-Cs-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C-C 6 -alkyl, heteroaryl-C 2
-C
6 alkenyl or heteroaryl-C 2
-C
6 -alkynyl, WO 2004/080480 PCT/DK2004/000158 65 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . Embodiment 86. A pharmaceutical composition according to embodiment 85 wherein R 24 ,
R
25 and R 26 are independently selected from 5 hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 ,
-NR
27
C(O)R
28 , -NR 2 7
C(O)OR
28 ,
-OC(O)R
2 7 , -OC 1
-C
6 -alkyl-C(O)OR 27 , -SC 1 -Ce-alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 2 7 _C -C 6 -alkyl-C(=O)OR 2 7, -C 1
-C
6 -alkyl-C(=O)OR 2 7 , or
-C(O)OR
27 , 10 * Cl-Ce-alkyl optionally substituted with one or more substituents independently se lected from R 29 * aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-C 1
-C
6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 15 alkyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . Embodiment 87. A pharmaceutical composition according to embodiment 86 wherein R 21 20 R 22 and R 23 are independently selected from * hydrogen, halogen, -OCF 3 , -OR 2 7 , -NR 2 7
R
2 8 , -SR 2 7 , -NR 27
C(O)R
28 , -NR 27
C(O)OR
2 8 ,
-OC(O)R
2 7 , -OCl-C 6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 2 7 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)N R 27 -Cl -C 6 -alkyl-C(=O)OR 2 7 , -CI-C 6 -alkyl-C(=O)OR 27 , or 25 -C(O)OR 2 , * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 30 *ArG1, ArG1-O-, ArG1-C(O)-, ArG1-Cl-C 6 -alkoxy, ArG1-C 1
-C
6 -alkyl, Het3, Het3-Cj
C
6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . Embodiment 88. A pharmaceutical composition according to embodiment 87 wherein R 21 , 35 R 22 and R 23 are independently selected from WO 2004/080480 PCT/DK2004/000158 66 * hydrogen, halogen, -OCFa, -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27
C(O)R
28 , -NR 27
C(O)OR
28 ,
-OC(O)R
27 , -OC 1 -C6-alkyl-C(O)OR 27 , -SCl-C 6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)NR 27
-C
1
-C
6 -alkyl-C(=O)OR 2 7 , -CI-C 6 -alkyl-C(=O)OR 27 , or 5 -C(O)OR 2 7 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 2 9 10 *ArG1, ArG1-O-, ArGI-C(O)-, ArG1-C-C6-alkoxy, ArG1-Cl-C 6 -alkyl, Het3, Het3-C
C
6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 ). Embodiment 89. A pharmaceutical composition according to embodiment 88 wherein R 21 , 15 R 22 and R 23 are independently selected from *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27
R
28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27
C(O)OR
2 8 ,
-OC(O)R
2 7 , -OCl-CB-alkyl-C(O)OR 2 7 , -SC 1
-C
6 -alkyl-C(O)OR 27 , -C 2
-C
6 -alkenyl
C(=O)OR
27 , -C(=O)N R 2 7 -Cl -Ce-alkyl-C(=O)OR 27 , -C 1
-C
6 -alkyl-C(=O)OR 27 , or 20 -C(O)OR 27 , * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 2 9 *ArG1, ArG1-O-, ArGl1-Cl-C 6 -alkoxy, ArGl-CI-C 6 -alkyl, 25 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30 . Embodiment 90. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 89 wherein R 20 is hydrogen or methyl. Embodiment 91. A pharmaceutical composition according to embodiment 90 wherein R 20 is 30 hydrogen. Embodiment 92. A pharmaceutical composition according to any one of the embodiment s60 or 78 to 91 wherein R 2 7 is hydrogen, Cl-C 6 -alkyl or aryl. Embodiment 93. A pharmaceutical composition according to embodiment 92 wherein R 27 is hydrogen or C 1
-C
6 -alkyl or ArG1.
WO 2004/080480 PCT/DK2004/000158 67 Embodiment 94. A pharmaceutical composition according to embodiment 93 wherein R 27 is hydrogen or Cl-Ce-alkyl. Embodiment 95. A pharmaceutical composition according to any one of the embodiment s 60 or 78 to 93 wherein R 28 is hydrogen or CI-C 6 -alkyl. 5 Embodiment 96. A pharmaceutical composition according to embodiment 60 wherein R" 7 and
R
18 are independently selected from *hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NOz, -OR 27 , -NR 27
R
28 , -SR 27 , -S(O)R 27 ,
-S(O)
2
R
27 , -C(O)N R 27
R
28 , -CH 2 0R 27 , -OC(O)R 27 , -OCl-C 6 -alkyl-C(O)OR 27 , -SC1-C 6 10 alkyl-C(O)OR, or -C(O)OR 7 , * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, optionally substituted with one or more substituents independently selected from R 29 15 *aryl, aryloxy, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-Cl-Co-alkyl, heteroaryl, heteroaryl-Cl-C6 alkyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 0 . 20 Embodiment 97. A pharmaceutical composition according to embodiment 96 wherein R1 7 and
R
18 are independently selected from *hydrogen, halogen, -CN, -CF3, -NO 2 , -OR, -NR 27
R
28 , or-C(O)OR 27 25 * Cl-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 2 *aryl, aryloxy, aroyl, aryl-C l
-C
6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C6 alkyl, 30 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 . Embodiment 98. A pharmaceutical composition according to embodiment 97 wherein R 17 and
R
18 are independently selected from 35 *hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 27
R
28 , or -C(O)OR 2 7 WO 2004/080480 PCT/DK2004/000158 68 . methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-C;-C6 alkyl 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 Embodiment 99. A pharmaceutical composition according to embodiment 98 wherein R' 7 and
R
18 are independently selected from *hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 2 7
R
28 , or -C(O)OR 27 10 methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 9 * ArG1, ArG1-O-, ArG1-C(O)-, ArG1-C 1
-C
6 -alkoxy, ArGl1-Cl-C 6 -alkyl, Het3, Het3-C
C
6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se 15 lected from R. Embodiment 100. A pharmaceutical composition according to embodiment 99 wherein R 17 and R' 8 are independently selected from * hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 2 7
R
28 , or -C(O)OR 27 * C 1
-C
6 -alkyl optionally substituted with one or more substituents independently se 20 lected from R 2 9 * phenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, phenyl-Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 . Embodiment 101. A pharmaceutical composition according to any one of the embodiment s 25 60 to 100 wherein R 27 is hydrogen or Cl-C 6 -alkyl. Embodiment 102. A pharmaceutical composition according to embodiment 101 wherein R 27 is hydrogen, methyl or ethyl. Embodiment 103. A pharmaceutical composition according to any one of the embodiment s 60 to 102 wherein R 28 is hydrogen or Cl-C 6 -alkyl. 30 Embodiment 104. A pharmaceutical composition according to embodiment 103 wherein R 2 " is hydrogen, methyl or ethyl. Embodiment 105. A pharmaceutical composition according to any one of the embodiment s 60 to 104 wherein R 72 is -OH or phenyl. Embodiment 106. A pharmaceutical composition according to embodiment 60 wherein CGr 35 is WO 2004/080480 PCT/DK2004/000158 69 N N H Embodiment 107. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein COr is of the form H-I-J 5 wherein H is 0 OH O OH HO or HO or HO N 10 H wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 15 I is selected from *a valence bond, * -CH 2
N(R
32 )- or -SO 2
N(R
33 )-, -ZL- N ]n * wherein Z' is S(O) 2 or CH 2 , Z 2 is -NH-, -O-or -S-, and n is 1 or 2, 20 J is * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R3 , *Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-Cl-C 6 -alkoxy-, aryl-C 1
-C
6 -alkyl-, aryl-C 2 C 6 -alkenyl-, aryl-C 2
-C
6 -alkynyl-, heteroaryl, heteroaryl-Cl-C 6 -alkyl-, heteroaryl-C 2
-C
6 25 alkenyl- or heteroaryl-C 2
-C
6 -alkynyl-, wherein the cyclic moieties are optionally substi tuted with one or more substituents selected from R 37 , * hydrogen,
R
3 1 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF3, -OCF 3 , 30 -OCHF 2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -S(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 3 5 , -C(O)R s , -NR 35
R
36 , -SR 3
,
WO 2004/080480 PCT/DK2004/000158 70 -NRaS(O) 2
R
36 , -S(O) 2
NR
5
R
36 , -S(O)NR 35
R
36 , -S(O)R 3 5 , -S(O) 2
R
35 , -C(O)NR 35
R
36 ,
-OC(O)NR"R
3 6 , -NR 35
C(O)R
3 , -CH 2
C(O)NR
35
R
3 8, -OCH 2
C(O)NRR
3 6 , -CH 2 0R 35 ,
-CH
2
NR
35
R
36 , -OC(O)R 5 , -OC,-C 6 -alkyl-C(O)OR 35 , -SCi-C 6 -alkyl-C(O)OR 35
-C
2
-C
6 -alkenyl
C(=O)OR
3 5 , -NR 3 s- 5
C(=O)-C
6 -C-alkyl-C(=O)OR 35 , -NR 3 -C(=O)-C-C-alkenyl-C(=O)OR 3 5 - , 5 C 1 -CB-alkyl, C 1
-C
6 -alkanoyl or -C(O)OR 35 , R1 2 and R 33 are independently selected from hydrogen, C 1
-C
6 -alkyl or C 1
-C
6 -alkanoyl,
R
34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR5R 3 , 10
R
35 and R 36 are independently selected from hydrogen, Cl-C6-alkyl, aryl-C 1 -CB-alkyl or aryl, or
R
3 5 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two 15 double bonds,
R
37 is independently selected from halogen, -C(O)OR 5 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , OR 3 5 , -NR 35
R
36 , C 1
-C
6 -alkyl or C 1
-C
6 -alkanoyl, 20 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 108. A pharmaceutical composition according to embodiment 107 wherein CGr is of the form H-1-J, wherein H is 0 HO OH HO OH Hor HO or HO HO ,o N H wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one 25 or more substituents independently selected from R 31 , I is selected from *a valence bond, * -CH2N(R32)- or -SOzN(R3)-, WO 2004/080480 PCT/DK2004/000158 71 -ZI-N' n Z2 * wherein Z' is S(O) 2 or CH 2 , Z 2 is N,-O-or -S-, and n is 1 or 2, J is * C 1 -Ce-alkyl, C2-C6-alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted 5 with one or more substituents selected from R3, *Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-Ci-C 6 -alkoxy-, aryl-C 1
-C
6 -alkyl-, aryl-C 2 C 6 -alkenyl-, aryl-C 2
-C
6 -alkynyl-, heteroaryl, heteroaryl-Cl-Ce-alkyl-, heteroaryl-C 2
-C
6 alkenyl- or heteroaryl-C 2
-C
6 -alkynyl-, wherein the cyclic moieties are optionally substi tuted with one or more substituents selected from R 37 , 10 e hydrogen,
R
3 1 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 ,
-OCHF
2 , -OCH 2
CF
3 , -OCF 2
CHF
2 , -S(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR s 35 , -C(O)R 3s , -NR R 3 ", -SR 3 ,
-NR
35
S(O)
2
R
36 , -S(O) 2
NR
3 5
R
3 6 , -S(O)NR 35
R
36 , -S(O)R 35 , -S(O) 2
R
35 , -C(O)NR 35 R 3 , 15 -OC(O)NR 35
R
36 , -NR 35
C(O)R
36 , -CH 2
C(O)NR
35
R
3 6 , -OCH 2
C(O)NR
35
R
36 , -CH 2 0R 35 ,
-CH
2
NR
5
R
36 , -OC(O)R 3 5 , -OC-C-alkyl-C(O)OR 3 5 , -SC 1
-C
6 -alkyl-C(O)OR 35
-C
2
-C
6 -alkenyl
C(=O)OR
35 , -NR 3 -C(=O)-Cs-C 6 -alkyl-C(=O)OR s , -NR 3 s-C(=O)-Cl-C 6 -alkenyl-C(=O)OR 35 -, Cl-C 6 -alkyl, Cl-Ce-alkanoyl or -C(O)OR, 20 R 32 and R 33 are independently selected from hydrogen, CI-C 6 -alkyl or Cl-C 6 -alkanoyl, R3 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR, and -NR 3 5
R
36 , R 35 and R 3 " are independently selected from hydrogen, C 1
-C
6 -alkyl, aryl-Cl-C 6 -alkyl or aryl, or 25 R 35 s and R 3 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 30 R 37 is independently selected from halogen, -C(O)OR 35 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , OR 35 , -NR 35
R
3 8 , C 1
-C
6 -alkyl or Cl-C 6 -alkanoyl, WO 2004/080480 PCT/DK2004/000158 72 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R 31 and J cannot both be hydrogen. Embodiment 109. A pharmaceutical composition according to any one of the embodiment s 5 107 or 108 wherein H is O 0 HOor Ho HO HO Embodiment 110. A pharmaceutical composition according to embodiment 109 wherein H is O HO HO Embodiment 111. A pharmaceutical composition according to embodiment 109 wherein H is 0 HO -' HO 10 Embodiment 112. A pharmaceutical composition according to any one of the embodiment s 107 to 111wherein I is a valence bond, -CH 2
N(R
32 )-, or-SO 2
N(R
3 3 )-. Embodiment 113. A pharmaceutical composition according to embodiment 112 wherein I is a valence bond. 15 Embodiment 114. A pharmaceutical composition according to any one of the embodiment s 107 to 113 wherein J is * hydrogen, * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents selected from 20 halogen, -CN, -CF 3 , -OCF 3 , -OR 3 5 , and -NR 35
R
36 , * aryl, or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 . Embodiment 115. A pharmaceutical composition according to embodiment 114 wherein J is * hydrogen, WO 2004/080480 PCT/DK2004/000158 73 earyl or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 . Embodiment 116. A pharmaceutical composition according to embodiment 114 wherein J is * hydrogen, 5 *ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 . Embodiment 117. A pharmaceutical composition according to embodiment 116 wherein J is * hydrogen, * phenyl or naphthyl optionally substituted with one or more substituents inde 10 pendently selected from R 37 . Embodiment 118. A pharmaceutical composition according to embodiment 117 wherein J is hydrogen. Embodiment 119. A pharmaceutical composition according to any one of the embodiment s 107 to 118 wherein R 32 and R 33 are independently selected from hydrogen or Cl-C 6 -alkyl. 15 Embodiment 120. A pharmaceutical composition according to any one of the embodiment s 107 to 119 wherein R3 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -OR 3 5 ,
-C(O)R
3 5 , -NR 35
R
36 , -SR 35 , -C(O)NR 35
R
3 6 , -OC(O)NR 3 5R 36 , -NR 35
C(O)R
3 6 , -OC(O)R 35 , -OC 1 C 6 -alkyl-C(O)OR 3 5 , -SCl-C 6 -alkyl-C(O)OR s or -C(O)OR 5 . Embodiment 121. A pharmaceutical composition according to embodiment 120 wherein R3 20 is hydrogen, halogen, -CF3, -NO 2 , -ORa, -NR 35
R
36 , -SR 5 , -NR 35
C(O)R
36 , or -C(O)OR3. Embodiment 122. A pharmaceutical composition according to embodiment 121 wherein R 34 is hydrogen, halogen, -CF3, -NO 2 , -ORa, -NR 35
R
36 , or -NR 3 5
C(O)R
36 . Embodiment 123. A pharmaceutical composition according to embodiment 122 wherein R3 is hydrogen, halogen, or -OR 5. 25 Embodiment 124. A pharmaceutical composition according to any one of the embodiment s 107 to 123 wherein R 35 and R 3 are independently selected from hydrogen, C 1
-C
6 -alkyl, or aryl. Embodiment 125. A pharmaceutical composition according to embodiment 124 wherein R 35 and R 36 are independently selected from hydrogen or CI-CB-alkyl. 30 Embodiment 126. A pharmaceutical composition according to any one of the embodiment s 107 to 125 wherein R 37 is halogen, -C(O)OR 35 , -CN, -CF 3 , -OR 35 , -NR 3 5
R
36 , Cl-C 6 -alkyl or Cl
C
6 -alkanoyl. Embodiment 127. A pharmaceutical composition according to embodiment 126 wherein R 37 is halogen, -C(O)ORa, -OR", -NR 35
R
36 , C 1
-C
6 -alkyl or C 1
-C
6 -alkanoyl.
WO 2004/080480 PCT/DK2004/000158 74 Embodiment 128. A pharmaceutical composition according to embodiment 127 wherein R 37 is halogen, -C(O)OR 35 or -OR 35 . Embodiment 129. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is /N HN -- ' K, \M___Q, \ / M N=N T wherein K is a valence bond, C 1
-C
6 -alkylene, -NH-C(=O)-U-, -CI-C 8 -alkyl-S-, -Cl-C 6 -alkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R, 10 U is a valence bond, Cl-C 6 -alkenylene, -Cl-C 6 -alkyl-O- or Cl-C 6 -alkylene wherein any C 1 C 6 -alkyl moiety is optionally substituted with Cl-C 6 -alkyl,
R
38 is Cl-C 6 -alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R 39 , 15
R
39 is independently selected from halogen, cyano, nitro, amino, M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 40 , 20
R
40 is selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 ,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 4 1 , -NR 4 1
R
42 , -SR 4 1
-NR
41
S(O)
2
R
4 2 , -S(0) 2
NR
4 1
R
42 , -S(O)NR 'R 42 , -S(O)R 41 , -S(O) 2
R
41 , -OS(0) 2
R
4 1 25 -C(O)NR 41
R
4 , -OC(O)NR 41
R
42 , -NR 41 C(O)R 42 , -CH 2
C(O)NR
4 1
R
42 , -OC-CB alkyl-C(O)NR 4 ' R 42 , -CH 2 0R 41 , -CH20C(O)R 41 , -CH 2
NR
41
R
42 , -OC(O)R 4 1, -OC-Ca alkyl-C(O)OR 41 , -OC-Cs-alkyl-OR 41 , -S-C 1
-C
5 -alkyl-C(O)OR 41 , -C 2
-C
6 -alkenyl
C(=O)OR
41 , -NR 4
'-C(=O)-C
1
-C
6 -alky)-C(=O)OR 41 , -NR 4 -C(=O)-Cl-C 6 alkenyl-C(=O)OR 41 , -C(O)OR 4 1 , -C 2
-C
6 -alkenyl-C(=O)R 41 , =O, -NH-C(=O)-O-Cl 30 C 6 -alkyl, or -NH-C(=O)-C(=O)-O-C-C 6 -alkyl, * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R 4
,
WO 2004/080480 PCT/DK2004/000158 75 *aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C 1 l-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C
C
6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl or heteroaryl-C 2
-C
6 -alkynyl, wherein the cyclic 5 moieties optionally may be substituted with one or more substituents selected from R",
R
4 1 and R 42 are independently selected from hydrogen, -OH, C 1
-C
6 -alkyl, CI-C 6 -alkenyl, aryl Cl-C 6 -alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more 10 substituents independently selected from R 45 , and the aryl moieties may optionally be substi tuted with one or more substituents independently selected from R 46 ; R 4 1 and R 42 when at tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double 15 bonds,
R
43 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 4 1 , and -NR 4 1
R
4 2 R4 is independently selected from halogen, -C(O)OR 41 , -CH 2
C(O)OR
41 , -CH 2 0R 4 1 , -CN, 20 CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 41
R
42 and Cl-Ce-alkyl,
R
4 5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -O-Cl-C 6 -alkyl, -C(O)-O-Cl
C
6 -alkyl, -COOH and -NH 2 ,
R
46 is independently selected from halogen, -C(O)OC,-C 6 -alkyl, -COOH, -CN, -CF3, -OCF 3 , NO 2 , -OH, -OCl-C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, 25 Q is a valence bond, C-C 6 -alkylene, -C 1
-C
6 -alkyl-O-, -C 1
-C
6 -alkyl-NH-, -NH-C l
-C
6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-C 1
-C
6 -alkyl, -C(=O)-, or -C 1
-C
6 -alkyl-C(=O)-N(R 4 7 )- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R 48 , 30
R
47 and R 48 are independently selected from hydrogen, C 1
-C
6 -alkyl, aryl optionally substituted with one or more R 49 ,
R
49 is independently selected from halogen and -COOH, 35 WO 2004/080480 PCT/DK2004/000158 76 T is * hydrogen, * CI-C6-alkyl, C 2 -Ce-alkenyl , C 2
-C
6 -alkynyl, C 1
-C
6 -alkyloxy-carbonyl, wherein the alkyl, 5 alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R 5 , *aryl, aryoxy, aryloxy-carbony), aryl-Cl-C 8 -alkyl, aroyl, aryl-C 1
-C
6 -alkoxy, aryl-C 2 C6-alkenyl, aryl-C 2
-C
6 -alkyny-, heteroaryl, heteroaryl-Cl-CB-alkyl, heteroaryl-C 2 C 6 -alkenyl, heteroaryl-Cz-C6-alkynyl, 10 wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 5 ,
R
5 0 is Cl-C 6 -alkyl, C -C 6 -alkoxy, aryl, aryloxy, aryl-Cl-C 6 -alkoxy, -C(=O)-NH-C,-C 6 -alkyl-aryl, 15 -C(=O)-NR50A-c1-C6-alkyl, -C(=O)-NH-(CH 2 CH20)mC-C 6 -alkyl-COOH, heteroaryl, het eroaryl-Cs-C 6 -alkoxy, -Cl-C 6 -alkyl-COOH, -O-Cl-C 6 -alkyl-COOH, -S(O) 2
R
1 ,
-C
2
-C
6 -alkenyl-COOH, -OR 5 , -NO 2 , halogen, -COOH, -CF 3 , -CN, =O, -N(Re'R 5 2 ), wherein m is 1, 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R 53 , and the alkyl moieties are optionally substituted with one or more R 5 OB. 20 Embodiment R50A and R51 are independently selected from -C(O)OCI-C 6 -alkyl, -COOH, -C
C
6 -alkyl-C(O)OC,-C 6 -alkyl, -CI-C 6 -alkyl-COOH, or C 1
-C
6 -alkyl,
R
5 ' and R 52 are independently selected from hydrogen and CI-C 5 -alkyl,
R
53 is independently selected from Cl-C 6 -alkyl, Cl-C 6 -alkoxy, -C,-C 5 -alkyl-COOH, -C2
C
6 -alkenyl-COOH, -OR 51 , -NO 2 , halogen, -COOH, -CF 3 , -ON, or -N(R 5 IR52), 25 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 130. A pharmaceutical composition according to embodiment 129 wherein K is a valence bond, C-C 6 -alkylene, -NH-C(=O)-U-, -Cs-C 6 -alkyl-S-, -C l
-C
6 -alkyl-O-, or -C(=O)-, 30 wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R 38 . Embodiment 131. A pharmaceutical composition according to embodiment 130 wherein K is a valence bond, C-C 6 -alkylene, -NH-C(=O)-U-, -C 1
-C
6 -alkyl-S-, or -C 1
-C
6 -alkyl-O, wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R 3
.
WO 2004/080480 PCT/DK2004/000158 77 Embodiment 132. A pharmaceutical composition according to embodiment 131 wherein K is a valence bond, CI-C 6 -alkylene, or -NH-C(=O)-U, wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R 3 8 . Embodiment 133. A pharmaceutical composition according to embodiment 132 wherein K is 5 a valence bond or Cl-C6-alkylene, wherein any C 1
-C
6 -alkyl moiety is optionally substituted with R 38 . Embodiment 134. A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or -NH-C(=O)-U. Embodiment 135. A pharmaceutical composition according to embodiment 133 wherein K is 10 a valence bond. Embodiment 136. A pharmaceutical composition according to any one of the embodiment s 129 to 135 wherein U is a valence bond or -C 1
-C
6 -alkyl-O-. Embodiment 137. A pharmaceutical composition according to embodiment 136 wherein U is a valence bond. 15 Embodiment 138. A pharmaceutical composition according to any one of the embodiment s 129 to 137 wherein M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from
R
40 Embodiment 139. A pharmaceutical composition according to embodiment 138 wherein M is 20 ArG1 or Hetl, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 4 . Embodiment 140. A pharmaceutical composition according to embodiment 139 wherein M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R4. 25 Embodiment 141. A pharmaceutical composition according to embodiment 140 wherein M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 4 . Embodiment 142. A pharmaceutical composition according to embodiment 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from 30 R 40 . Embodiment 143. A pharmaceutical composition according to embodiment 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from
R
4 0 Embodiment 144. A pharmaceutical composition according to embodiment 143 wherein M is WO 2004/080480 PCT/DK2004/000158 78
R
4 0 145. A pharmaceutical composition according to embodiment 141 wherein M is carbazolylene optionally substituted with one or more substituents independently selected from R. 5 Embodiment 146. A pharmaceutical composition according to embodiment 145 wherein M is
R
4 0 Embodiment 147. A pharmaceutical composition according to any one of the embodiment s 129 to 146 wherein R 4 0 is selected from hydrogen, halogen, -CN, -CF3, -OCF 3 , -NO 2 , -OR 41 , -NR 4 1R 42 , -SR 4 1 , -S(O) 2
R
41 ', 10 -NR 41
C(O)R
42 , -OCI-C 6 -alkyl-C(0)NR 41
R
42 , -C 2 -C6-alkenyl-C(=O)OR 41 , -C(O)OR 41 =0, -NH-C(=O)-O-Cs-C 6 -alkyl, or -NH-C(=O)-C(=O)-O-C 1
-C
6 -alkyl, Cl-C 6 -alkyl or C 2
-C
6 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 15 *oaryl, aryloxy, aryl-C 1
-C
6 -alkoxy, aryl-Cl-CB-alkyl, aryl-C 2
-C
6 -alkenyl, heteroaryl, het eroaryl-Cl-C 6 -alkyl, or heteroaryl-C 2
-C
6 -alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R". Embodiment 148. A pharmaceutical composition according to embodiment 147 wherein R 4 0 20 is selected from *hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 41
R
42 , -SR 41 , -S(0) 2
R
41 ,
-NR
41
C(O)R
4 2 , -OC 1
-C
6 -alkyl-C(0)NR 41
R
42 , -C 2
-C
6 -alkenyl-C(=O)OR 1 , -C(0)OR 41 =O, -NH-C(=O)-O-CI-C 6 -alkyl, or -NH-C(=O)-C(=O)-O-Cj-C 6 -alkyl, 25 Cl-C 6 -alkyl or C 2
-C
6 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 , *ArG1, ArG1-O-, ArGl-Cl-Ce-alkoxy, ArGl-C 1
-C
6 -alkyl, ArG1-C 2
-C
6 -alkenyl, Het3, Het3-Cl-C 6 -alkyl, or Het3-C 2
-C
6 -alkenyl, wherein the cyclic moieties optionally may be 30 substituted with one or more substituents selected from R 44
".
WO 2004/080480 PCT/DK2004/000158 79 Embodiment 149. A pharmaceutical composition according to embodiment 148 wherein R 4 0 is selected from *hydrogen, halogen, -CF 3 , -NO 2 , -OR 41 , -NR 41
R
4 2 , -C(O)OR 41 , =0, or -NR41C(O)R 42 , * C 1 -CB-alkyl, 5 ArG1. Embodiment 150. A pharmaceutical composition according to embodiment 149 wherein R 40 is hydrogen. Embodiment 151. A pharmaceutical composition according to embodiment 149 wherein R 4 is selected from 10 *halogen, -NO 2 , -OR 41 , -NR 4 1
R
42 , -C(O)OR 41 , or -NR 41C (
O)R
42 , * methyl, Sphenyl. Embodiment 152. A pharmaceutical composition according to any one of the embodiment s 129 to 151 wherein R 41 and R 42 are independently selected from hydrogen, Cl-C 6 -alkyl, or 15 aryl, wherein the aryl moieties may optionally be substituted with halogen or-COOH. Embodiment 153. A pharmaceutical composition according to embodiment 152 wherein R 4 1 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH. Embodiment 154. A pharmaceutical composition according to any one of the embodiment s 20 129 to 153 wherein Q is a valence bond, Cl-C 6 -alkylene, -Cl-C 6 -alkyl-0-, -Cj-C 6 -alkyl-NH-, -NH-Cl-C 6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-Cl-C 6 -alkyl, -C(=O)-, or -C,
C
6 -alkyl-C(=O)-N(R7)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R 48 Embodiment 155. A pharmaceutical composition according to embodiment 154 wherein Q is 25 a valence bond, -CH 2 -, -CH 2
-CH
2 -, -CH 2 -O-, -CH 2
-CH
2 -O-, -CH 2 -NH-, -CH 2
-CH
2 -NH-,
-NH-CH
2 -, -NH-CH 2
-CH
2 -, -NH-C(=O)-, -C(=O)-NH-, -O-CH 2 -, -O-CH 2
-CH
2 -, or -C(=O)-. Embodiment 156. A pharmaceutical composition according to any one of the embodiment s 129 to 155 wherein R 47 and R 48 are independently selected from hydrogen, methyl and phenyl. 30 Embodiment 157. A pharmaceutical composition according to any one of the embodiment s 129 to 156 wherein T is * hydrogen, * C,-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 5
,
WO 2004/080480 PCT/DK2004/000158 80 earyl, aryl-Cl-C 6 -alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 5 0 . Embodiment 158. A pharmaceutical composition according to embodiment 157 wherein T is * hydrogen, 5 * C-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 50 , *ArG1, ArG1-C 1
-C
6 -alkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are op tionally substituted with one or more substituents independently selected from R 5 0 . Embodiment 159. A pharmaceutical composition according to embodiment 158 wherein T is 10 * hydrogen, * Cl-C 6 -alkyl, optionally substituted with one or more substituents independently se lected from R 5 , ephenyl, phenyl-Cl-C 6 -alkyl, wherein the alkyl and phenyl moieties are optionally substituted with one or more substituents independently selected from R". 15 Embodiment 160. A pharmaceutical composition according to embodiment 159 wherein T is phenyl substituted with R 5 . Embodiment 161. A pharmaceutical composition according to any one of the embodiment s 129 to 160 wherein R 50 is CI-C 6 -alkyl, Cl-C 6 -alkoxy, aryl, aryloxy, aryl-C 1
-C
6 -alkoxy,
-C(=O)-NH-C,-C
6 -alkyl-aryl, -C(=O)-NRs50A 1 c -C 6 -alkyl, -C(=O)-NH-(CH 2
CH
2 0)mCl-C 6 -alkyl 20 COOH, heteroaryl, -Cl-C 6 -alkyl-COOH, -O-Cl-C 6 -alkyl-COOH, -S(O)2R 51
-C
2
-C
6 -alkenyl-COOH, -OR 5 1 , -NO 2 , halogen, -COOH, -CF 3 , -CN, =0, -N(R 51
R
5 2 ), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R 5 3 . Embodiment 162. A pharmaceutical composition according to embodiment 161 wherein R 50 is Cl-C 6 -alkyl, Cl-C 6 -alkoxy, aryl, aryloxy, -C(=O)-NR50A-C1-C6-alkyl, 25 -C(=O)-NH-(CH 2
CH
2 0)mCi-C6-alkyl-COOH, aryl-Cl-C 6 -alkoxy , -OR 5 1 , -NO 2 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 5 3 . Embodiment 163. A pharmaceutical composition according to embodiment 162 wherein R5 is Cl-C 6 -alkyl, aryloxy, -C(=O)-NRs0AC1 -C 6 -alkyl, -C(=O)-NH-(CH 2
CH
2 0)mCi-C 6 -alkyl-COOH, aryl-Cl-C 6 -alkoxy, -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substi 30 tuted with one or more R3. Embodiment 164. A pharmaceutical composition according to embodiment 163 wherein R 5 0 is Cl-C 6 -alkyl, ArG1-O-, -C(=O)-NR50A ci-C6-alkyl, -C(=O)-NH-(CH2
C
H20)mC,-C 6 -alkyl COOH, ArG1-Cl-C 6 -alkoxy , -OR 5 1 , halogen, -COOH, -CF 3 , wherein any aryl moiety is op tionally substituted with one or more R5.
WO 2004/080480 PCT/DK2004/000158 81 Embodiment 165. A pharmaceutical composition according to embodiment 164 wherein R" is -C(=O)-NR50A CH 2 , -C(=O)-NH-(CH 2
CH
2 0) 2
CH
2 1-COOH, or -C(=O)-NR50A CH 2
CH
2 . Embodiment 166. A pharmaceutical composition according to embodiment 164 wherein R s 50 is phenyl, methyl or ethyl. 5 Embodiment 167. A pharmaceutical composition according to embodiment 166 wherein R 5 0 is methyl or ethyl. Embodiment 168. A pharmaceutical composition according to any one of the embodiment s 129 to 167 wherein m is 1 or 2. Embodiment 169. A pharmaceutical composition according to any one of the embodiment s 10 129 to 168 wherein R 51 is methyl. Embodiment 170. A pharmaceutical composition according to any one of the embodiment s 129 to 169 wherein R 53 is Ci-C 6 -alkyl, Co-Co-alkoxy, -OR 51 , halogen,or -CF 3 . Embodiment 171. A pharmaceutical composition according to any one of the embodiment s 129 to 170 wherein R 5 OA is -C(O)OCH 3 , -C(O)OCH 2
CH
3 -COOH, -CH 2
C(O)OCH
3 , 15 CH 2
C(O)OCH
2
CH
3 , -CH 2
CH
2
C(O)OCH
3 , -CH 2
CH
2
C(O)OCH
2
CH
3 , -CH2COOH, methyl, or ethyl. Embodiment 172. A pharmaceutical composition according to any one of the embodiment s 129 to 171 wherein RO s B is -C(O)OCH 3 , -C(O)OCH 2
CH
3 -COOH, -CH 2
C(O)OCH
3 , CH 2
C(O)OCH
2
CH
3 , -CH 2
CH
2
C(O)OCH
3 , -CH 2
CH
2
C(O)OCH
2
CH
3 , -CH 2 COOH, methyl, or 20 ethyl. Embodiment 173. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is N'N HN' S 25 wherein V is Cl-Co-alkyl, aryl, heteroaryl, aryl-Cs 6 -alkyl- or aryl-C 2 -6-alkenyl-, wherein the al kyl or alkenyl is optionally substituted with one or more substituents independently selected from R4, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 5 , 30 R' is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
R
s5 is independently selected from WO 2004/080480 PCT/DK2004/000158 82 *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCHZCFa,
-OCF
2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 8 , -NRseR 5 7 , -SR 5 ,
-NR
56
S(O)
2
R
57 , -S(O) 2
NR"R
, 7 , -S(O)NRR 57 , -S(O)R 56 , -S(O) 2
R
56 , -OS(O) 2 Rm, -C(O)NR R s 7 , -OC(O)N R 56
R
57 , -NR 56
C(O)R
6 7 , -CH 2
C(O)NR
56
R
57 , -OC 1
-C
6 5 alkyl-C(O)NR"R 5 7 , -CH 2 0R 5 , -CH 2
OC(O)R
s6 , -CH 2
NRR
57 , -OC(O)R 5 , -OC-C alkyl-C(O)OR 6 , -OC-Ce-alkyl-OR 5 ", -SC.-Co-alkyl-C(O)OR 5 0, -C 2
-C
6 -alkenyl C(=O)OR, -NR 5 6-C(=O)-Cl-C 6 -alkyl-C(=O)OR 56, -NR6-C(=O)-C 1 l-C 6 alkenyl-C(=O)OR 5 6 , -C(O)OR 5 6 , or -C 2 -CB-alkenyl-C(=O)R 6 , 10 * C 1 -C6-alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, which may optionally be substituted with one or more substituents selected from R 58 , earyl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C,-C 6 -alkoxy, aryl-C 1
-C
6 -alkyl, 15 aryl-C2-C 6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C,
C
6 -alkyl, heteroaryl-C 2 -C6-alkenyl or heteroaryl-C 2
-C
6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R s , 20
R
56 and R 57 are independently selected from hydrogen, OH, CF3,; C-C 1 2 -alkyl, aryl-CI-C alkyl, -C(=O)-Cj-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 6 0 , and the aryl groups may option ally be substituted with one or more substituents independently selected from R 6 ; R 5 6 and 25 R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 30 R5 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 5 6 , and -NR 56
R
57 ,
R
59 is independently selected from halogen, -C(O)OR s 56 , -CH 2
C(O)OR
56 , -CH 2 0R 56 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR 56 , -NR 56
R
57 and Cl-C 6 -alkyl, WO 2004/080480 PCT/DK2004/000158 83
R
6 o is independently selected from halogen, -CN, -CF3, -OCF 3 , -OC,-C-alkyl, -C(O)OC 1
-C
6 alkyl, -C(=O)-R 62 , -COOH and -NH 2 ,
R
61 is independently selected from halogen, -C(O)OC-C6-alkyl, -COOH, -CN, -CF 3 , -OCF 3 , 5 NO 2 , -OH, -OCl-C 6 -alkyl, -NH 2 , C(=O) or CI-C 6 -alkyl,
R
6 2 is C,-C 6 -alkyl, aryl optionally substituted with one or more substituents independently se lected from halogen, or heteroaryl optionally substituted with one or more Cl-C 6 -alkyl inde pendently, 10 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 174. A pharmaceutical composition according to embodiment 173 wherein V is aryl, heteroaryl, or aryl-C 1 .e-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R5, and the aryl or heteroaryl is optionally substituted 15 with one or more substituents independently selected from R 5 5 . Embodiment 175. A pharmaceutical composition according to embodiment 174 wherein V is aryl, Hetl, or aryl-C 1 .e-alkyl-, wherein the alkyl is optionally substituted with one or more sub stituents independently selected from R5, and the aryl or heteroaryl moiety is optionally sub stituted with one or more substituents independently selected from R 55 . 20 Embodiment 176. A pharmaceutical composition according to embodiment 175 wherein V is aryl, Het2, or aryl-C l .6-alkyl-, wherein the alkyl is optionally substituted with one or more sub stituents independently selected from R", and the aryl or heteroaryl moiety is optionally sub stituted with one or more substituents independently selected from R' 5 . Embodiment 177. A pharmaceutical composition according to embodiment 176 wherein V is 25 aryl, Het3, or aryl-C 1 6-alkyl-, wherein the alkyl is optionally substituted with one or more sub stituents independently selected from R5, and the aryl or heteroaryl moiety is optionally sub stituted with one or more substituents independently selected from R 5 5 . Embodiment 178. A pharmaceutical composition according to embodiment 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R 55 . 30 Embodiment 179. A pharmaceutical composition according to embodiment 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R 55 s . Embodiment 180. A pharmaceutical composition according to embodiment 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independ ently selected from R ss 5 5
.
WO 2004/080480 PCT/DK2004/000158 84 Embodiment 181. A pharmaceutical composition according to embodiment 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R 5 5 . Embodiment 182. A pharmaceutical composition according to any one of the embodiment s 173 to 181 wherein R 55 is independently selected from 5 *halogen, Cl-Cs-alkyl, -CN, -OCF 3
,-CF
3 , -NO 2 , -OR 5 6 , -NR 56
R
5 7 , -NR 56
C(O)R
57
-SR
5 6 , -OC 1 -Ce-alkyl-C(O)OR 56 , or -C(O)OR", * C,-Cs-alkyl optionally substituted with one or more substituents independently se lected from R9 8 * aryl, aryl-Cl-C 6 -alkyl, heteroaryl, or heteroaryl-C,-C 6 -alkyl 10 of which the cyclic moieties optionally may be substituted with one or more substitu ents independently selected from R 59 . Embodiment 183. A pharmaceutical composition according to embodiment 182 wherein R' 5 is independently selected from *halogen, C,-Co-alkyl, -CN, -OCF 3
,-CF
3 , -NO 2 , -OR ' 6 , -NR6R 57 , -NR5"C(O)R 5 7 15 -SR 56 , -OC 1
-C
8 -alkyl-C(O)ORm, or -C(O)OR 5 6 * C,-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 58 * ArG1, ArGl-Cl-C 5 -alkyl, Het3, or Het3-C,-C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substitu 20 ents independently selected from R 5 9 . Embodiment 184. A pharmaceutical composition according to embodiment 183 wherein R 55 is independently selected from halogen, -OR 6 , -NR 56 Rs 7 , -C(O)OR 1 6 , -OC 1
-C
8 alkyl-C(O)OR 5 6 , -NR 56
C(O)R
57 or Cl-C 6 -alkyl. Embodiment 185. A pharmaceutical composition according to embodiment 184 wherein R s 5 25 is independently selected from halogen, -OR 56 , -NR 56
R
57 , -C(O)OR 5 6 , -OC 1 -C8 alkyl-C(O)OR", -NR"C(O)R 57 , methyl or ethyl. Embodiment 186. A pharmaceutical composition according to any one of the embodiment s 173 to 185 wherein R 56 and R 57 are independently selected from hydrogen, CF 3 , C 1
-C
1 2 -alkyl, or -C(=O)-C 1
-C
6 -alkyl; R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 30 8 membered heterocyclic ring with the said nitrogen atom. Embodiment 187. A pharmaceutical composition according to embodiment 186 wherein Ree and R 5 7 are independently selected from hydrogen or C-C 12 -alkyl, R 56 and R 57 when at tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
WO 2004/080480 PCT/DK2004/000158 85 Embodiment 188. A pharmaceutical composition according to embodiment 187 wherein R s 56 and R 5 7 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. 5 Embodiment 189. A pharmaceutical composition according to any one of the embodiment s 1 to 3 wherein CGr is N H N N AA 10 wherein AA is Cl-C 6 -alkyl, aryl, heteroaryl, aryl-Cl 6 -alkyl- or aryl-C 2
.
6 -alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 64 15 R 63 is independently selected from halogen, -CN, -CF3, -OCF 3 , aryl, -COOH and -NH 2 ,
R
64 is independently selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2
CF
3 , 20 -OCF 2
CHF
2 , -S(O) 2
CF
3 , -OS(O) 2
CF
3 , -SCF 3 , -NO 2 , -OR 65 es , -NR 6
*R
66 , -SR 6 5,
-NR
6 5
S(O)
2
R
66 , -S(0) 2
NR
65
R
66 , -S(O)NR 5
R
6 6 , -S(O)R 6 5 , -S(O) 2
R
65 , -OS(0) 2
R
6 5 , -C(O)NRe 6
R
66 , -OC(O)NR 6 5
R
66 , -NR 6 sC(O)R 66 , -CH 2 C(O)NRe 65
R
66 , -OCs-C6 alkyl-C(O)NR 65
R
66 , -CH 2 0R 65 , -CH 2
OC(O)R
6 5 , -CH 2
NR
65
R
66 , -OC(O)R 65 s , -OCo-Cs alkyl-C(O)OR 65 , -OC,-C 6 -alkyl-OR 65 , -SC-C6-alkyl-C(O)OR 65 , -C 2
-C
6 -alkenyl 25 C(=O)OR 6 5 , -NR 6 5 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 6 5 , -NR 65 -C(=O)-Cl-C alkenyl-C(=O)OR 6 5 , -C(O)OR 6 5 , or -C 2
-C
6 -alkenyl-C(=O)R 65 , * Cl-C 6 -alkyl, C 2
-C
6 -alkenyl or C 2
-C
6 -alkynyl, each of which may optionally be substi tuted with one or more substituents selected from R1 7 , 30 WO 2004/080480 PCT/DK2004/000158 86 Saryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-C 1
-C
6 -alkyl, aryl-C 2
-C
6 -alkenyl, aroyl-C 2
-C
6 -alkenyl, aryl-C 2
-C
6 -alkynyl, heteroaryl, heteroaryl-C
C
6 -alkyl, heteroaryl-C 2
-C
6 -alkenyl or heteroaryl-C 2 -C6-alkynyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 68 ,
R
65 and R 66 are independently selected from hydrogen, OH, CF 3 , Cl-C 1 2 -alkyl, aryl-C 1
-C
6 alkyl, -C(=O)-R 69 , aryl or heteroaryl, wherein the alkyl groups may optionally be substituted 10 with one or more substituents selected from R 7 0 , and the aryl and heteroaryl groups may op tionally be substituted with one or more substituents independently selected from R 71 ; R 65 and R 66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or 15 two double bonds,
R
67 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 6 5 , and -NR 65
R
66 ,
R
68 is independently selected from halogen, -C(O)OR 6 5 , -CH 2
C(O)OR
65 , -CH 2 0R 65 , -CN, 20 CF 3 , -OCF 3 , -NO 2 , -OR" 5 , -NR 65
R
6 6 and Cl-C 6 -alkyl,
R
6g is independently selected from CI-C 5 -alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more Cl-C 6 -alkyl, 25 R 7 0 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OC 1
-C
6 -alkyl, -C(O)OCI-C6 alkyl, -COOH and -NH 2 ,
R
71 is independently selected from halogen, -C(O)OC-C 6 -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , NO 2 , -OH, -OC-C 6 -alkyl, -NH 2 , C(=O) or C 1
-C
6 -alkyl, 30 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a-salt thereof with a pharmaceutically acceptable acid or base. Embodiment 190. A pharmaceutical composition according to embodiment 189 wherein AA is aryl, heteroaryl or aryl-Cl6-alkyl-, wherein the alkyl is optionally substituted with one or WO 2004/080480 PCT/DK2004/000158 87 more R 6 3 , and the aryl or heteroaryl is optionally substituted with one or more substitLuents independently selected from R6. Embodiment 191. A pharmaceutical composition according to embodiment 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently se 5 lected from R 64. Embodiment 192. A pharmaceutical composition according to embodiment 191 wherein AA is ArG1 or Hetl optionally substituted with one or more substituents independently selected from R 64 . Embodiment 193. A pharmaceutical composition according to embodiment 192 wherein AA 10 is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R6. Embodiment 194. A pharmaceutical composition according to embodiment 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 6 " . 15 Embodiment 195. A pharmaceutical composition according to embodiment 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R 64 Embodiment 196. A pharmaceutical composition according to embodiment 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently se 20 lected from R6. Embodiment 197. A pharmaceutical composition according to any one of the embodiment s 189 to 196 wherein R 6 4 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 ,
-OR
65 , -NR 5
R
66 , Ci-C 6 -alkyl , -OC(O)R 65 , -OC 1
-C
6 -alkyl-C(O)OR 6 5 , aryl-C 2
-C
6 -alkenyl, ary loxy or aryl, wherein C 1
-C
6 -alkyl is optionally substituted with one or more substituents inde 25 pendently selected from R 67 , and the cyclic moieties optionally are substituted with one or more substituents independently selected from R 68 . Embodiment 198. A pharmaceutical composition according to embodiment 197 wherein R " is independently selected from halogen, -CF 3 , -OCF 3 , -OR 6 5 , -NRs 65
R
66 , methyl, ethyl, propyl,
-OC(O)R
6 5 , -OCH 2
-C(O)OR
6 5 , -OCH 2
-CH
2
-C(O)OR
6 5 , phenoxy optionally substituted with one 30 or more substituents independently selected from R 8 . Embodiment 199. A pharmaceutical composition according to any one of the embodiment s 189 to 198 wherein R 65 and R 6 6 are independently selected from hydrogen, CF 3 , C,-C 1 2 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently se lected from R 71
.
WO 2004/080480 PCT/DK2004/000158 88 Embodiment 200. A pharmaceutical composition according to embodiment 199 whereinr R 6 5 and R 66 are independently hydrogen, C 1
-C
1 2 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R 71 . Embodiment 201. A pharmaceutical composition according to embodiment 200 wherein R 6 * 5 5 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Hetl optionally substituted with one or more substituents independently selected from R 71 Embodiment 202. A pharmaceutical composition according to embodiment 201 wherein R 65 and R 6 6 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 71 . 10 Embodiment 203. A pharmaceutical composition according to embodiment 202 wherein R 6 5 and R 6 6 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 71 . Embodiment 204. A pharmaceutical composition according to embodiment 203 wherein R 6 5 and R 6 6 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, 15 naphtyl, thiadiazolyl optionally substituted with one or more R 71 independently; or isoxazolyl optionally substituted with one or more substituents independently selected from R 71 . Embodiment 205. A pharmaceutical composition according to any one of the embodiment s 189 to 204 wherein R 71 is halogen or C 1
-C
6 -alkyl. Embodiment 206. A pharmaceutical composition according to embodiment 205 wherein R 71 20 is halogen or methyl. Embodiment 207. A pharmaceutical preparation according to any one of the embodiment s 1 to 205 wherein FrgI consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. Embodiment 208. A pharmaceutical preparation according to embodiment 207 wherein Frgl 25 consists of 0 to 5 Gly. Embodiment 209. A pharmaceutical preparation according to embodiment 208 wherein Frgl consists of 0 Gly. Embodiment 210. A pharmaceutical preparation according to embodiment 208 wherein Frgl consists of 1 Gly. 30 Embodiment 211. A pharmaceutical preparation according to embodiment 208 wherein Frgl consists of 2 Gly. Embodiment 212. A pharmaceutical preparation according to embodiment 208 wherein Frgl consists of 3 Gly. Embodiment 213. A pharmaceutical preparation according to embodiment 208 wherein Frgl 35 consists of 4 Gly.
WO 2004/080480 PCT/DK2004/000158 89 Embodiment 214. A pharmaceutical preparation according to embodiment 208 wherein Frgl consists of 5 Gly. Embodiment 215. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein GE is of the formula B'-B 2 -C(O)-, B 1
-B
2
-SO
2 - or B'-B 2
-CH
2 -, wherein B 1 and 5 B 2 are as defined in embodiment 1. Embodiment 216. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G is of the formula B'-B 2 -C(O)-, B 1
-B
2
-SO
2 - or B 1
-B
2 -NH-, wherein B and B 2 are as defined in embodiment 1. Embodiment 217. A pharmaceutical preparation according to any one of the embodiment s 1 10 to 214 wherein G B is of the formula B'-B 2 -C(O)-, B 1
-B
2
-CH
2 - or B 1
-B
2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1. Embodiment 218. A pharmaceutical preparation according to any one of the embodiment s 1 to 214 wherein G B is of the formula B 1
-B
2
-CH
2 -, B'-B 2
-SO
2 - or B 1 -B 2 -NH-, wherein B1 and B 2 are as defined in embodiment 1. 15 Embodiment 219. A pharmaceutical preparation according to any one of the embodiment s 215 or 216 wherein GB is of the formula B 1
-B
2 -C(O)- or B'-B 2
-SO
2 -, wherein B 1 and B 2 are as defined in embodiment 1. Embodiment 220. A pharmaceutical preparation according to any one of the embodiment s 215 or 217 wherein GB is of the formula B 1
-B
2 -C(O)- or B'-B 2
-CH
2 -, wherein B 31 and B 2 are as 20 defined in embodiment 1. Embodiment 221. A pharmaceutical preparation according to any one of the embodiment s 216 or 217 wherein GB is of the formula B'-B -C(O)- or B'-B 2 -NH-, wherein B 1 and B2 are as defined in embodiment 1. Embodiment 222. A pharmaceutical preparation according to any one of the embodiment s 25 215 or 218 wherein GB is of the formula B'-B 2
-CH
2 - or B'-B 2 -SO2-, wherein B 1 and B 2 are as defined in embodiment 1. Embodiment 223. A pharmaceutical preparation according to any one of the embodiment s 216 or 218 wherein G3 is of the formula B 1
-B
2 -NH- or B3 1 -B 2
-SO
2 - , wherein B1 and B 2 are as defined in embodiment 1. 30 Embodiment 224. A pharmaceutical preparation according to any one of the embodiment s 217 or 218 wherein G B is of the formula B'-B 2
-CH
2 - or B 1 -B -NH- , wherein B' and B2 are as defined in embodiment 1. Embodiment 225. A pharmaceutical preparation according to any one of the embodiment s 219, 220, or 221 wherein GB is of the formula B'-B 2
-C(O)-.
WO 2004/080480 PCT/DK2004/000158 90 Embodiment 226. A pharmaceutical preparation according to any one of the embodiment s 220, 222 or 224 wherein GB is of the formula B 1
-B
2
-CH
2 -. Embodiment 227. A pharmaceutical preparation according to any one of the embodiment s 220, 222 or 223 wherein GB is of the formula B 1
-B
2
-SO
2 -. 5 Embodiment 228. A pharmaceutical preparation according to any one of the embodiment s 221, 223 or 224 wherein GB is of the formula B'-B 2 -NH-. Embodiment 229. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is a valence bond, -0-, or -S-. Embodiment 230. A pharmaceutical preparation according to any one of the embodiment s 1 10 to 228 wherein B 1 is a valence bond, -0-, or -N(R 6 B)-. Embodiment 231. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is a valence bond, -S-, or -N(R 6 B)-. Embodiment 232. A pharmaceutical preparation according to any one of the embodiment s 1 to 228 wherein B 1 is -0-, -S- or -N(R6B)-. 15 Embodiment 233. A pharmaceutical preparation according to any one of the embodiment s 229 or 230 wherein B' is a valence bond or -0-. Embodiment 234. A pharmaceutical preparation according to any one of the embodiment s 229 or 231 wherein B' is a valence bond or -S-. Embodiment 235. A pharmaceutical preparation according to any one of the embodiment s 20 230 or 231 wherein B' is a valence bond or -N(R 6 ")-. Embodiment 236. A pharmaceutical preparation according to any one of the embodiment s 229 or 232 wherein B' is -O-or -S-. Embodiment 237. A pharmaceutical preparation according to any one of the embodiment s 230 or 232 wherein B 1 is -O-or -N(R 6 B)-. 25 Embodiment 238. A pharmaceutical preparation according to any one of the embodiment s 231 or 232 wherein B 1 is -S-or -N(R6B)-. Embodiment 239. A pharmaceutical preparation according to any one of the embodiment s 233, 234 or 235 wherein B 1 is a valence bond. Embodiment 240. A pharmaceutical preparation according to any one of the embodiment s 30 233, 236 or 237 wherein B 1 is -0-. Embodiment 241. A pharmaceutical preparation according to any one of the embodiment s 234, 236 or 238 wherein B' is -S-. Embodiment 242. A pharmaceutical preparation according to any one of the embodiment s 235, 237 or 238 wherein B' is -N(R 6
B)_.
WO 2004/080480 PCT/DK2004/000158 91 Embodiment 243. A pharmaceutical preparation according to any one of the embodiment s 1 to 242 wherein B 2 is a valence bond, C 1
-C
1 8 -alkylene, C 2
-C
1 8 -alkenylene, C 2
-C
18 -alkynylene, arylene, heteroarylene, -C l
-C
18 -alkyl-aryl-, -C(=O)-C 1
-C
18 -alkyI-C(=O)-, -C(=O)-C 1
-C
1 8 -alkyl O-C-Cj-alkyl-C(=O)-, -C(=O)-Cl-C 1 8 -alkyl-S-C-C 8 -alkyl-C(=O)-, -C(=O)-C 1
-C
18 -alkyl-NR 5 Cl-C, 8 -alkyl-C(=O)-; and the alkylene and arylene moieties are optionally substituted as de fined in embodiment 1. Embodiment 244. A pharmaceutical preparation according to embodiment 243 wherein B 2 is a valence bond, Cl-C 1 8 -alkylene, C 2 -Cl8-alkenylene, C 2
-C
1 8 -alkynylene, arylene, heteroary lene, -Cl-C 18 -alkyl-aryl-, -C(=O)-C-C 18 -alkyl-C(=O)-, -C(=O)-Cl-C 18 -alkyl-O-Cl-Cs-alkyl 10 C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in em bodiment 1. Embodiment 245. A pharmaceutical preparation according to embodiment 244 wherein B 2 is a valence bond, Cl-C 1 8 -alkylene, C 2
-C
18 -alkenylene, C 2
-C,
8 -alkynylene, arylene, heteroary lene, -C 1
-C
1 8-alkyl-aryl-, -C(=O)-Cl-Cj 8 -alkyl-C(=O)-, and the alkylene and arylene moieties 15 are optionally substituted as defined in embodiment 1. Embodiment 246. A pharmaceutical preparation according to embodiment 245 wherein B 2 is a valence bond, Cl-C, 8 -alkylene, arylene, heteroarylene, -Cl-C, 8 -alkyl-aryl-, -C(=O)-C-C 8 alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1. 20 Embodiment 247. A pharmaceutical preparation according to embodiment 246 wherein B 2 is a valence bond, C 1
-C
18 -alkylene, arylene, heteroarylene, -C 1
-C
18 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1. Embodiment 248. A pharmaceutical preparation according to embodiment 247 wherein B 2 is a valence bond, C 1
-C
1 8 -alkylene, arylene, -Cl-C 8 -alkyl-aryl-, and the alkylene and arylene 25 moieties are optionally substituted as defined in embodiment 1. Embodiment 249. A pharmaceutical preparation according to embodiment 248 wherein B 2 is a valence bond or -Cl-C, 8 -alkylene, and the alkylene moieties are optionally substituted as defined in embodiment 1. Embodiment 250. A pharmaceutical preparation according to any one of the embodiment s 1 30 to 249 whereiin Frg2 comprises 1 to 16 positively charged groups. Embodiment 251. A pharmaceutical preparation according to embodiment 250 wherein Frg2 comprises 1 to 12 positively charged groups. Embodiment 252. A pharmaceutical preparation according to embodiment 251 wherein Frg2 comprises 1 to 10 positively charged groups.
WO 2004/080480 PCT/DK2004/000158 92 Embodiment 253. A pharmaceutical preparation according to any one of the embodiment s 1 to 249 wherein Frg2 comprises 10 to 20 positively charged groups. Embodiment 254. A pharmaceutical preparation according to embodiment 253 wherein Frg2 comprises 12 to 20 positively charged groups. 5 Embodiment 255. A pharmaceutical preparation according to embodiment 254 wherein Frg2 comprises 16 to 20 positively charged groups. Embodiment 256. A pharmaceutical preparation according to any one of the embodiment s 250 to 255 wherein the positively charged groups of Frg2 are basic amino acids independ ently selected from the group consisting of Lys and Arg and D-isomers of these. 10 Embodiment 257. A pharmaceutical preparation according to embodiment 256 wherein the basic amino acids are all Arg. Embodiment 258. A pharmaceutical preparation according to any one of the embodiment s 250 to 257, wherein Frg2 comprises one or more neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser. 15 Embodiment 259. A pharmaceutical preparation according to embodiment 258, wherein Frg2 comprises one or more Gly. Embodiment 260. A pharmaceutical preparation according to any one of the embodiment s 1 to 259 wherein X is -OH or -NH 2 . Embodiment 261. A pharmaceutical preparation according to embodiment 260 wherein X is 20 NH 2 . Embodiment 262. A pharmaceutical preparation according to any one of the embodiment s 1 to 261 which further comprises at least 3 phenolic molecules per putative insulin hexamer. Embodiment 263. A pharmaceutical preparation according to any one of the embodiment s 1 to 262 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, 25 Gin, Glu, Gly, His, lie, Leu, Met, Phe, Ser, Thr, Trp, Tyr, Val, and hSer. Embodiment 264. A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gly, lie, Leu, Phe, Ser, Thr, Val, and hSer. Embodiment 265. A pharmaceutical preparation according to embodiment 264 wherein the 30 acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala or Gly. Embodiment 266. A pharmaceutical preparation according to embodiment 265 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Gly. Embodiment 267. A pharmaceutical preparation according to any one of the embodiment s 263 to 266 wherein the acid-stabilised insulin is an analogue of human insulin further modi 35 fied by exchange or deletion of one or more amino acid residues according to the following: WO 2004/080480 PCT/DK2004/000158 93 B3 is selected from Thr, Ser, Lys or Ala A18 is Gin B28 is Lys, Asp or Glu B29 is Pro or Glu 5 B9 is Glu or Asp B10 is Glu B25 is deleted B30 is deleted. Embodiment 268. A pharmaceutical preparation according to embodiment 267 wherein the 10 acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys or Asp. Embodiment 269. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Asp. 15 Embodiment 270. A pharmaceutical preparation according to embodiment 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys. Embodiment 271. A pharmaceutical preparation according any one of the embodiment s 263 to 270 wherein the acid-stabilised insulin is an analogue of human insulin further modified by 20 exchange of B29 to Pro. Embodiment 272. A pharmaceutical preparation according any one of the embodiment s 263 to 271 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B3 to Lys or Ala. Embodiment 273. A pharmaceutical preparation according any one of the embodiment s 263 25 to 272 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of A18 to Gin. Embodiment 274. A pharmaceutical preparation according any one of the embodiment s 263 to 273 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B25. 30 Embodiment 275. A pharmaceutical preparation according any one of the embodiment s 263 to 274 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B30. Embodiment 276. A pharmaceutical preparation according to embodiment 263 wherein the acid-stabilised insulin is selected from the group 35 A21G WO 2004/080480 PCT/DK2004/000158 94 A21G, B28K, B29P A21G, B28D A21G, B28E A21G, B3K, B29E 5 A21G,desB27 A21G, B9E A21G, B9D A21G, B10E A21G, desB25 10 A21G,desB30 A21G, B28K, B29P, desB30 A21G, B28D, desB30 A21G, B28E, desB30 A21G, B3K, B29E, desB30 15 A21G, desB27, desB30 A21G, B9E, desB30 A21G, B9D, desB30 A21 G, B10OE, desB30 A21G, desB25, desB30. 20 Embodiment 277. A pharmaceutical preparation according to any one of the embodiment s 1 to 276 wherein zinc ions are present in an amount corresponding to 10 to 40 gg Zn/100 U insu lin. Embodiment 278. A pharmaceutical preparation according to embodiment 277 wherein zinc ions are present in an amount corresponding to 10 to 26 pg Zn/100 U insulin. 25 Embodiment 279. A pharmaceutical preparation according to any one of the embodiment s 1 to 278 wherein the ratio between insulin and the zinc-binding ligand according to any one of the embodiment s 1 to 261 is in the range from 99:1 to 1:99. Embodiment 280. A pharmaceutical preparation according to embodiment 279 wherein the ratio between insulin and the zinc-binding ligand according to any one of the embodiment s 1 30 to 261 is in the range from 95:5 to 5:95. Embodiment 281. A pharmaceutical preparation according to embodiment 280 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the em bodiment s 1 to 261 is in the range from 80:20 to 20:80.
WO 2004/080480 PCT/DK2004/000158 95 Embodiment 282. A pharmaceutical preparation according to embodiment 281 wherein the ratio between between insulin and the zinc-binding ligand according to any one of the em bodiment s 1 to 261 is in the range from 70:30 to 30:70. Embodiment 283. A pharmaceutical preparation according to any one of the embodiment s 1 5 to 282 wherein the concentration of insulin is 60 to 3000 nmol/ml. Embodiment 284. A pharmaceutical preparation according to embodiment 283 wherein the concentration of insulin is 240 to 1200 nmol/ml. Embodiment 285. A pharmaceutical preparation according to embodiment 284 wherein the concentration of insulin is about 600 nmol/ml. 10 Embodiment 286. A method of preparing a zinc-binding ligand according to embodiment 1 comprising the steps of * Identifying starter compounds that binds to the R-state HisBlo-Zn 2 + site . optionally attaching a fragment consisting of 0 to 5 neutral a- or 13-amino acids * attaching a fragment comprising 1 to 20 positively charged groups independently se 15 lected from amino or guanidino groups. Embodiment 287. Method of prolonging the action of an acid-stabilised insulin preparation which comprises adding a zinc-binding ligand according to any of embodiment s 1 to 261 to the acid-stabilised insulin preparation. Embodiment 288. A method of treating type 1 or type 2 diabetes comprising administering to 20 a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the embodiment s 1 to 282. Embodiment 289. Use of a preparation according to any one of the embodiment s 1 to 282 for the preparation of a medicament for treatment of type 1 or type 2 diabetes. 25 PHARMACEUTICAL PREPARATIONS The present invention also relates to a pharmaceutical preparation for the treatment of diabe tes in a patient in need of such a treatment comprising an R-state hexamer of insulin accord ing to the invention together with a pharmaceutically acceptable carrier. 30 In one embodiment of the invention the insulin preparation comprises 60 to 3000 nmol/ml of in sulin. In another embodiment of the invention the insulin preparation comprises 240 to 1200 nmol/ml of insulin.
WO 2004/080480 PCT/DK2004/000158 96 In another embodiment of the invention the insulin preparation comprises about 600 nmol/rnml of insulin. Zinc ions may be present in an amount corresponding to 10 to 40 lag Zn/1 00 U insulin, more preferably 10 to 26 lig Zn/100 U insulin. 5 Insulin formulations of the invention are usually administered from multi-dose containers where a preservative effect is desired. Since phenolic preservatives also stabilize the R-state hexamer the formulations may contain up to 50 mM of phenolic molecules. The phenolic molecules in the insulin formulation may be selected from the group consisting of phenol, m cresol, chloro-cresol, thymol, 7-hydroxyindole or any mixture thereof. 10 In one embodiment of the invention 0.5 to 4.0 mg/ml of phenolic compound may be employed. In another embodiment of the invention 0.6 to 4.0 mg/ml of m-cresol may be employed. In another embodiment of the invention 0.5 to 4.0 mg/ml of phenol may be employed. In another embodiment of the invention 1.4 to 4.0 mg/ml of phenol may be employed. In another embodiment of the invention 0.5 to 4.0 mg/ml of a mixture of m-cresol or phenol 15 may be employed. In another embodiment of the invention 1.4 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed. The pharmaceutical preparation may further comprises a buffer substance, such as a TRIS, phosphate, glycine or glycylglycine (or another zwitterionic substance) buffer, an isotonicity 20 agent, such as NaCI, glycerol, mannitol and/or lactose. Chloride would be used at moderate concentrations (e.g. up to 50 mM) to avoid competition with the zinc-site ligands of the pre sent invention. The action of insulin may further be slowed down in vivo by the addition of physiologically acceptable agents that increase the viscosity of the pharmaceutical preparation. Thus, the 25 pharmaceutical preparation according to the invention may furthermore comprise an agent which increases the viscosity, such as polyethylene glycol, polypropylene glycol, copolymers thereof, dextrans and/or polylactides. In a particular embodiment the insulin preparation of the invention comprises between 0.001 % by weight and 1 % by weight of a non-ionic surfactant, for example tween 20 or Polox 188. 30 A nonionic detergent can be added to stabilise insulin against fibrillation during storage and handling. The insulin preparation of the present invention may have a pH value in the range of 2.5 to 4.5, more preferably pH 3 to 4.
WO 2004/080480 PCT/DK2004/000158 97 In one embodiment the preparations of the invention are used in connection with insulin pumps. The insulin pumps may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Insulin pumps may be skin-mounted or car ried, and the path of the insulin preparation from the storage compartment of the pump to the 5 patient may be more or less tortuous. Non-limiting examples of insulin pumps are disclosed in US 5,957,895, US 5,858,001, US 4,468,221, US 4,468,221, US 5,957,895, US 5,858,001, US 6,074,369, US 5,858,001, US 5,527,288, and US 6,074,369. In another embodiment the preparations of the invention are used in connection with pen-like 10 injection devices, which may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Non-limiting examples of pen-like injection de vices are FlexPen®, InnoLeto, InDuoTM, Innovo
®
. In a further embodiment preparations of the invention are used in connection with devices for 15 pulmonary administration of aqueous insulin preparations, a non-limiting example of which is the AerX8 device. COMBINATION TREATMENT 20 The invention furthermore relates to treatment of a patient in which the pharmaceutical preparation of the invention, i.e. comprising zinc ions, acid-stabilised insulin analogue and a ligand for the R-state His B10 Zn 2 + site, is combined with another form of treatment. In one aspect of the invention, treatment of a patient with the pharmaceutical prepa ration of the invention is combined with diet and/or exercise. 25 In another aspect of the invention the pharmaceutical preparation of the invention is administered in combination with one or more further active substances in any suitable ratios. Such further active substances may e.g. be selected from antiobesity agents, antidiabetics, antihypertensive agents, agents for the treatment of complications resulting from or associ ated with diabetes and agents for the treatment of complications and disorders resulting from 30 or associated with obesity. Thus, in a further aspect of the invention the pharmaceutical preparation of the in vention may be administered in combination with one or more antiobesity agents or appetite regulating agents.
WO 2004/080480 PCT/DK2004/000158 98 Such agents may be selected from the group consisting of CART (cocaine am phetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melano cortin 4) agonists, MC3 (melanocortin 3) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releas 5 ing factor binding protein) antagonists, urocortin agonists, p3 adrenergic agonists such as CL-316243, AJ-9677, GW-0604, LY362884, LY377267 or AZ-40140, MSH (melanocyte stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake inhibitors such as fluoxetine, seroxat or cita lopram, serotonin and noradrenaline re-uptake inhibitors, mixed serotonin and noradrenergic 10 compounds, 5HT (serotonin) agonists, bombesin agonists, galanin antagonists, growth hor mone, growth factors such as prolactin or placental lactogen, growth hormone releasing compounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin agonists, DA agonists (bromocriptin, doprexin), lipase/amylase in hibitors, PPAR (peroxisome proliferator-activated receptor) modulators, RXR (retinoid X re 15 ceptor) modulators, TR P agonists, AGRP (Agouti related protein) inhibitors, H3 histamine antagonists, opioid antagonists (such as naltrexone), exendin-4, GLP-1 and ciliary neurotro phic factor. In one embodiment of the invention the antiobesity agent is leptin. In another embodiment the antiobesity agent is dexamphetamine or amphetamine. 20 In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine. In still another embodiment the antiobesity agent is sibutramine. In a further embodiment the antiobesity agent is orlistat. In another embodiment the antiobesity agent is mazindol or phentermine. In still another embodiment the antiobesity agent is phendimetrazine, diethylpropion, 25 fluoxetine, bupropion, topiramate or ecopipam. The orally active hypoglycemic agents comprise imidazolines, sulphonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, insulin sensitizers, insulin secretagogues such as glimepride, a.-glucosidase inhibitors, agents acting on the ATP 30 dependent potassium channel of the p-cells eg potassium channel openers such as those disclosed in WO 97/26265, WO 99/03861 and WO 00/37474 (Novo Nordisk A/S) which are incorporated herein by reference, or mitiglinide, or a potassium channel blocker, such as BTS-67582, nateglinide, glucagon antagonists such as those disclosed in WO 99/01423 and WO 00/39088 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorpo 35 rated herein by reference, GLP-1 agonists such as those disclosed in WO 00/42026 (Novo WO 2004/080480 PCT/DK2004/000158 99 Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorporated herein by refer ence, DPP-IV (dipeptidyl peptidase-IV) inhibitors, PTPase (protein tyrosine phosphatase) in hibitors, inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or gly cogenolysis, glucose uptake modulators, GSK-3 (glycogen synthase kinase-3) inhibitors, 5 compounds modifying the lipid metabolism such as antilipidemic agents, compounds lower ing food intake, PPAR (peroxisome proliferator-activated receptor) and RXR (retinoid X re ceptor) agonists, such as ALRT-268, LG-1268 or LG-1069. In a further embodiment of the invention the pharmaceutical preparation of the in vention is administered in combination with a sulphonylurea e.g. tolbutamide, chlorpropa 10 mide, tolazamide, glibenclamide, glipizide, glimepiride, glicazide or glyburide. In another embodiment of the invention the pharmaceutical preparation of the inven tion is administered in combination with a biguanide, e.g. metformin. In yet another embodiment of the invention the pharmaceutical preparation of the in vention is administered in combination with a meglitinide eg repaglinide or nateglinide. 15 In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a thiazolidinedione insulin sensitizer, e.g. trogli tazone, ciglitazone, pioglitazone, rosiglitazone, isaglitazone, darglitazone, englitazone, CS 011/CI-1 037 or T 174 or the compounds disclosed in WO 97/41097, WO 97/41119, WO 97/41120, WO 00/41121 and WO 98/45292 (Dr. Reddy's Research Foundation), which are 20 incorporated herein by reference. In still another embodiment of the invention the pharmaceutical preparation of the invention may be administered in combination with an insulin sensitizer, e.g. such as GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297, GW-409544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516 or the compounds disclosed in 25 WO 99/19313, WO 00150414, WO 00/63191, WO 00/63192, WO 00/63193 (Dr. Reddy's Re search Foundation) and WO 00/23425, WO 00/23415, WO 00/23451, WO 00/23445, WO 00/23417, WO 00/23416, WO 00/63153, WO 00/63196, WO 00/63209, WO 00/63190 and WO 00/63189 (Novo Nordisk A/S), which are incorporated herein by reference. In a further embodiment of the invention the pharmaceutical preparation of the in 30 vention is administered in combination with an cL-glucosidase inhibitor, e.g. voglibose, ernigli tate, miglitol or acarbose. In another embodiment of the invention the pharmaceutical preparation of the inven tion is administered in combination with an agent acting on the ATP-dependent potassium channel of the p-cells, e.g. tolbutamide, glibenclamide, glipizide, glicazide, BTS-67582 or re 35 paglinide.
WO 2004/080480 PCT/DK2004/000158 100 In yet another embodiment of the invention the pharmaceutical preparation of the in vention may be administered in combination with nateglinide. In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with an antilipidemic agent, e.g. cholestyramine, 5 colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine. In another aspect of the invention, the pharmaceutical preparation of the invention is administered in combination with more than one of the above-mentioned compounds, e.g. in combination with metformin and a sulphonylurea such as glyburide; a sulphonylurea and 10 acarbose; nateglinide and metformin; acarbose and metformin; a sulphonylurea, metformin and troglitazone; metformin and a sulphonylurea; etc. Furthermore, the pharmaceutical preparation of the invention may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents are P-blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and 15 metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and a blockers such as doxazosin, urapidil, prazosin and terazosin. The pharmaceutical prepara tion of the invention may also be combined with NEP inhibitors such as candoxatril. 20 Further reference can be made to Remington: The Science and Practice of Phar macy, 1 9 th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995. It should be understood that any suitable combination of the compounds according to the invention with diet and/or exercise, one or more of the above-mentioned compounds and optionally one or more other active substances are considered to be within the scope of 25 the present invention. EXAMPLES The following examples and general procedures refer to intermediate compounds and final 30 products identified in the specification and in the synthesis schemes. The preparation of the compounds of the present invention is described in detail using the following examples, but the chemical reactions described are disclosed in terms of their general applicability to the preparation of compounds of the invention. Occasionally, the reaction may not be applicable as described to each compound included within the disclosed scope of the invention. The WO 2004/080480 PCT/DK2004/000158 101 compounds for which this occurs will be readily recognised by those skilled in the art. In these cases the reactions can be successfully performed by conventional modifications known to those skilled in the art, that is, by appropriate protection of interfering groups, by changing to other conventional reagents, or by routine modification of reaction conditions. 5 Alternatively, other reactions disclosed herein or otherwise conventional will be applicable to the preparation of the corresponding compounds of the invention. In all preparative methods, all starting materials are known or may easily be prepared from known starting materials. All temperatures are set forth in degrees Celsius and unless otherwise indicated, all parts and percentages are by weight when referring to yields and all parts are by volume when refer 10 ring to solvents and eluents. HPLC-MS (Method A) The following instrumentation was used: * Hewlett Packard series 1100 G1312A Bin Pump 15 * Hewlett Packard series 1100 Column compartment * Hewlett Packard series 1100 G13 15A DAD diode array detector * Hewlett Packard series 1100 MSD The instrument was controlled by HP Chemstation software. 20 The HPLC pump was connected to two eluent reservoirs containing: A: 0.01% TFA in water B: 0.01% TFA in acetonitrile The analysis was performed at 40 OC by injecting an appropriate volume of the sample (pref erably 1 fL) onto the column, which was eluted with a gradient of acetonitrile.
WO 2004/080480 PCT/DK2004/000158 102 The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table. Column Waters Xterra MS C-18 X 3 mm id Gradient 10% - 100% acetonitrile lineary during 7.5 min at 1.0 mL/min Detection UV: 210 nm (analog output from DAD) MS lonisation mode: API-ES Scan 100-1000 amu step 0.1 amu 5 HPLC-MS (Method B) The following instrumentation was used: Sciex API 100 Single quadropole mass spectrometer Perkin Elmer Series 200 Quard pump Perkin Elmer Series 200 autosampler 10 Applied Biosystems 785A UV detector Sedex 55 evaporative light scattering detector A Valco column switch with a Valco actuator controlled by timed events from the pump. The Sciex Sample control software running on a Macintosh PowerPC 7200 computer was 15 used for the instrument control and data acquisition. The HPLC pump was connected to four eluent reservoirs containing: A: Acetonitrile B: Water C: 0.5% TFA in water D: 0.02 M ammonium acetate The requirements for samples are that they contain approximately 500 pIg/mL of the com 20 pound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.) The analysis was performed at room temperature by injecting 20 giL of the sample solution 25 on the column, which was eluted with a gradient of acetonitrile in either 0.05% TFA or 0.002 WO 2004/080480 PCT/DK2004/000158 103 M ammonium acetate. Depending on the analysis method varying elution conditions were used. The eluate from the column was passed through a flow splitting T-connector, which passed 5 approximately 20 pL/min through approx. 1 m. 75 pi fused silica capillary to the API interface of API 100 spectrometer. The remaining 1.48 mL/min was passed through the UV detector and to the ELS detector. 10 During the LC-analysis the detection data were acquired concurrently from the mass spec trometer, the UV detector and the ELS detector. The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following table. 15 Column YMC ODS-A 120A s - 5p 3 mm x 50 mm id Gradient 5% - 90% acetonitrile in 0.05% TFA linearly during 7.5 min at 1.5 mL/min Detection UV: 214 nm ELS: 40 0 C MS Experiment: Start: 100 amu Stop: 800 amu Step: 0.2 amu Dwell: 0.571 msec Method: Scan 284 times = 9.5 min HPLC-MS (Method C) The following instrumentation is used: * Hewlett Packard series 1100 G1312A Bin Pump * Hewlett Packard series 1100 Column compartment 20 * Hewlett Packard series 1100 G1315A DAD diode array detector * Hewlett Packard series 1100 MSD * Sedere 75 Evaporative Light Scattering detector The instrument is controlled by HP Chemstation software. The HPLC pump is connected to two eluent reservoirs containing: A 0.01% TFA in water B 0.01% TFA in acetonitrile 25 WO 2004/080480 PCT/DK2004/000158 104 The analysis is performed at 40 0C by injecting an appropriate volume of the sample (pref erably 1 pl) onto the column which is eluted with a gradient of acetonitrile. The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table. Column Waters Xterra MS C-18 X 3 mm id 5pm Gradient 5% - 100% acetonitrile linear during 7.5 min at 1.5 ml/min Detection 210 nm (analogue output from DAD) ELS (analogue output from ELS) MS ionisation mode API-ES Scan 100-1000 amu step 0.1 amu 5 After the DAD the flow is divided yielding approximately 1 ml/min to the ELS and 0.5 ml/min to the MS. HPLC-MS (Method D) 10 The following instrumentation was used: Sciex API 150 Single Quadropole mass spectrometer Hewlett Packard Series 1100 G1312A Bin pump Gilson 215 micro injector Hewlett Packard Series 1100 G1315A DAD diode array detector 15 Sedex 55 evaporative light scattering detector A Valco column switch with a Valco actuator controlled by timed events from the pump. The Sciex Sample control software running on a Macintosh Power G3 computer was used for the instrument control and data acquisition. The HPLC pump was connected to two eluent reservoirs containing: A: Acetonitrile containing 0.05% TFA B: Water containing 0.05% TFA 20 The requirements for the samples are that they contain approximately 500 pg/ml of the com pound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.) WO 2004/080480 PCT/DK2004/000158 105 The analysis was performed at room temperature by injecting 20 .l of the sample solution on the column, which was eluted with a gradient of acetonitrile in 0.05% TFA The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 pl/min through approx. 1 m 75 g fused silica capillary to the API interface of 5 API 150 spectrometer. The remaining 1.48 ml/min was passed through the UV detector and to the ELS detector. During the LC-analysis the detection data were acquired concurrently from the mass spec trometer, the UV detector and the ELS detector. The LC conditions, detector settings and mass spectrometer settings used for the different 10 methods are given in the following table. Column Waters X-terra C18 5p 3 mm x 50 mm id Gradient 5% - 90% acetonitrile in 0.05% TFA linearly during 7.5 min at 1.5 ml/min Detection UV: 214 nm ELS: 40 0 C MS Experiment: Start: 100 amu Stop: 800 amu Step: 0.2 amu Dwell: 0.571 msec Method: Scan 284 times = 9.5 min WO 2004/080480 PCT/DK2004/000158 106 EXAMPLES 0102-0000-0273Example 1 HBOL 1H-Benzotriazole N -0 N H 5 010 2 -0000-0274Example 2 HBOL 5,6-Dimethyl-1 H-benzotriazole N, !CH 3 N\ N CH H 3 10 0 1 0 2 -0000-0275Example 3 HBOL 1H-Benzotriazole-5-carboxylic acid H ,N N OH 010 2 -0000-0280Example 4 HBOL 15 4-Nitro-1H-benzotriazole H N Np O O 0102-0000-0284Example 5 HBOL 5-Amino-1H-benzotriazole N
NH
2 N" N 20
H
WO 2004/080480 PCT/DK2004/000158 107 0102-0000-0287Example 6 HBOL 5-Chloro-1 H-benzotriazole N Cl N H 5 0102-0000-0286Example 7 HBOL 5-Nitro-1 H-benzotriazole H N N NO I O 0102-0000-3015Example 8 PEM 10 4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid H NI "N N, 0 OH O 0 4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid methyl ester (5.2 g, 17.6 mmol) was dissolved in THF (60 mL) and methanol (10 mL) was added followed by 1N sodium hydrox ide (35 mL). The mixture was stirred at room temperature for 16 hours and then 1N hydro 15 chloric acid (45 mL) was added. The mixture was added water (200 mL) and extracted with ethyl acetate (2 x 500 mL). The combined organic phases were evaporated in vacuo to afford 0.44 g of 4-[(1H-benzotriazole-5-carbonyl)amino]benzoic acid. By filtration of the aqueous phase a further crop of 4-[(1H-benzotriazole-5-carbonyl)amino]benzoic acid was isolated (0.52 g). 20 1 H-NMR (DMSO-d 6 ): 67.97 (4H, s), 8.03 (2H, m), 8.66 (1H, bs), 10.7 (1H, s), 12.6 (1H, bs); HPLC-MS (Method A): m/z: 283 (M+1); Rt = 1.85 min.
WO 2004/080480 PCT/DK2004/000158 108 General procedure (A) for preparation of compounds of general formula 11: 0 0 U N OH H N E H N + HN E N 119 Nil N :01: R H H 11 wherein D, E and R 1 9 are as defined above, and E is optionally substituted with up to three substituents R 21 , R 22 and R 23 independently as defined above. 5 The carboxylic acid of 1 H-benzotriazole-5-carboxylic acid is activated, ie the OH functionality is converted into a leaving group L (selected from eg fluorine, chlorine, bromine, iodine, 1 imidazolyl, 1,2,4-triazolyl, 1-benzotriazolyloxy, 1-(4-aza benzotriazolyl)oxy, pentafluoro phenoxy, N-succinyloxy 3,4-dihydro-4-oxo-3-(1,2,3-benzotriazinyl)oxy, benzotriazole 5-COO, 10 or any other leaving group known to act as a leaving group in acylation reactions. The acti vated benzotriazole-5-carboxylic acid is then reacted with R 2
-(CH
2 )n-B ' in the presence of a base. The base can be either absent (i.e. R 2
-(CH
2 )n-B' acts as a base) or triethylamine, N ethyl-N,N.-diisopropylamine, N-methylmorpholine, 2,6-lutidine, 2,2,6,6-tetramethylpiperidine, potassium carbonate, sodium carbonate, caesium carbonate or any other base known to be 15 useful in acylation reactions. The reaction is performed in a solvent solvent such as THF, di oxane, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reaction is performed between 0 oC and 80 oC, preferably between 20 oC and 40 oC. When the acylation is complete, the product is isolated by extraction, filtration, chromatography or other methods known to those skilled in the art. 20 The general procedure (A) is further illustrated in the following example: 0102-0000-1020Example 9 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid phenylamide H ,N N I H N N 0 25 Benzotriazole-5-carboxylic acid (856 mg), HOAt (715 mg) and EDAC (1.00 g) were dissolved in DMF (17.5 mL) and the mixture was stirred at room temperature 1 hour. A 0.5 mL aliqot of this mixture was added to aniline (13.7 pL, 0.15 mmol) and the resulting mixture was vigor ously shaken at room temperature for 16 hours. 1N hydrochloric acid (2 mL) and ethyl ace tate (1 mL) were added and the mixture was vigorously shaken at room temperature for 2 WO 2004/080480 PCT/DK2004/000158 109 hours. The organic phase was isolated and concentrated in vacuo to afford the title com pound. HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min. 5 The compounds in the following examples were similarly made. Optionally, the compounds may be isolated by filtration or by chromatography. 0102-0000-1019Example 10 (General Procedure (A))PEM 10 1H-Benzotriazole-5-carboxylic acid (4-methoxyphenyl)amide H HPLC-MS (Method A): m/z: 269 (M+1) & 291 (M+23); Rt = 2.41 min HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min. 15 0102-0000-1021 Example 11 (General Procedure (A))PEM (4-[(1H-Benzotriazole-5-carbonyl)amino]phenyl}carbamic acid tert-butyl ester H NI H H 0 CH 3 HPLC-MS (Method B): m/z: 354 (M+1); Rt = 4.58 min. 20 0102-0000-1022Example 12 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (4-acetylaminophenyl)amide H N "N Na N kCH, H HPLC-MS (Method B): m/z: 296 (M+1); Rt = 3.32 min. 25 0102-0000-1023Example 13 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (3-fluorophenyl)amide WO 2004/080480 PCT/DK2004/000158 110 H NI H - N F HPLC-MS (Method B): m/z: 257 (M+1); Rt = 4.33 min. 0102-0000-1024Example 14 (General Procedure (A))PEM 5 1H-Benzotriazole-5-carboxylic acid (2-chlorophenyl)amide H H CI "N 0- N HPLC-MS (Method B): m/z: 273 (M+1); Rt = 4.18 min. 0102-0000-1025Example 15 (General Procedure (A))PEM 10 4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid methyl ester H NI N 0 N , , O c H 3 0 HPLC-MS (Method A):m/z: 297 (M+1); Rt: 2,60 min. HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.30 min. 0102-0000-1026Example 16 (General Procedure (A))PEM 15 1H-Benzotriazole-5-carboxylic acid (4-butylphenyl)amide H N 0 HPLC-MS (Method B): m/z: 295 (M+1); Rt = 5.80 min. 0102-0000-1027Example 17 (General Procedure (A))PEM 20 1H-Benzotriazole-5-carboxylic acid (1-phenylethyl)amide H ,N N O
CH
3 HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.08 min.
WO 2004/080480 PCT/DK2004/000158 111 0102-0000-1028Example 18 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid benzylamide H IN N 0 HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.88 min. 5 0102-0000-1029Example 19 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 4-chlorobenzylamide H N D'... _ aj d ,H H y C N N 0 HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.40 min. 10 0102-0000-1030Example 20 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid 2-chlorobenzylamide H N . N' H o ci O CI HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.25 min. 15 0102-0000-1031Example 21 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 4-methoxybenzylamide N I H - O ' CH, N N . 0 HPLC-MS (Method B): m/z: 283 (M+1); Rt = 3.93 min. 20 0102-0000-1032Example 22 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 3-methoxybenzylamide N N o CH, 0 HPLC-MS (Method B): m/z: 283 (M+1); Rt = 3.97 min. 25 WO 2004/080480 PCT/DK2004/000158 112 0102-0000-1033Example 23 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (1,2-diphenylethyl)amide N'h H HPLC-MS (Method B): m/z: 343 (M+I); Rt = 5.05 min. 5 0102-0000-1034Example 24 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 3-bromobenzylamide H N N N Br O HPLC-MS (Method B): m/z: 331 (M+1); Rt = 4.45 min. 10 0102-0000-1035Example 25 (General Procedure (A))PEM 4-{[(1 H-Benzotriazole-5-carbonyl)amino]methyl}benzoic acid 0 o HPLC-MS (Method B): m/z: 297 (M+1); Rt = 3.35 min. 15 0102-0000-1036Example 26 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid phenethylamide H NH N N v HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.08 min. 20 0102-0000-1037Example 27 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(4-chlorophenyl)ethyl]amide H N HPLC-MS (Method 13): m/z: 301 (M+1); Rt = 4.50 rain.
WO 2004/080480 PCT/DK2004/000158 113 0102-0000-1038Example 28 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(4-methoxyphenyl)ethyl]amide N N N OCH, 5 HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.15 min. 0102-0000-1039Example 29 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(3-methoxyphenyl)ethyl]amide o 10 HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.13 min. 0102-0000-1040Example 30 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(3-chlorophenyl)ethyl]amide H N H N N CI 0 N 15 HPLC-MS (Method B): mlz: 301 (M+I); Rt = 4.55 min. 0102-0000-1041 Example 31 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (2,2-diphenylethyl)amide H N "N N N :C N 20 HPLC-MS (Method B): m/z: 343 (M+1); Rt = 5.00 min. 0102-0000-1042Example 32 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid (3,4-dichlorophenyl)methylamide H ,N H3 "N - N ci 0 "N ci WO 2004/080480 PCT/DK2004/000158 114 HPLC-MS (Method B): m/z: 321 (M+1); Rt = 4.67 min. 0102-0000-1043Example 33 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid methylphenylamide H N IQ C 0 N0: 5 HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.82 min. 0102-0000-1044Example 34 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid benzylmethylamide H N N,-, 10 o HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.05 min. 0102-0000-1045Example 35 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(3-chloro-4-methoxyphenyl)ethyl]methyl-amide H N5 0 HPLC-MS (Method B): m/z: 345 (M+1); Rt = 4.37 min. 0102-0000-1046Example 36 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid methylphenethylamide H N, N 20 HPLC-MS (Method B): m/z: 281 (M+1); Rt = 4.15 min. 0102-0000-1047Example 37 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(3,4-dimethoxyphenyl)ethyl]methylamide WO 2004/080480 PCT/DK2004/000158 115 H N I H H "N -r N_- 0X 0 N .CH 3 HPLC-MS (Method B): m/z: 341 (M+1); Rt = 3.78 min; 0102-0000-1048Example 38 (General Procedure (A))PEM 5 1H-Benzotriazole-5-carboxylic acid (2-hydroxy-2-phenylethyl)methylamide HPLC-MS (Method B): m/z: 297 (M+1); Rt = 3.48 min. Example 39 (General procedure (A)) 10 1H-Benzotriazole-5-carboxylic acid (3-bromophenyl)amide 0 N 'Ne NN Br NI H N H HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.19 min. Example 40 (General procedure (A)) 15 1H-Benzotriazole-5-carboxylic acid (4-bromophenyl)amide O Br N N N N N I H H HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.18 min. Example 41 (General procedure (A)) 20 {4-[(1H-Benzotriazole-5-carbonyl)amino]benzoylamino}acetic acid _~ ~ ~ .N--,TrOH HOH , N NO H 0 N 'N H H HPLC-MS (Method A): m/z: 340 (M+1); Rt = 1.71 min.
WO 2004/080480 PCT/DK2004/000158 116 Example 42 (General procedure (A)) {4-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}acetic acid N OH *N H H 5 HPLC-MS (Method A): m/z: 297 (M+1); Rt = 2.02 min. Example 43 (General procedure (A)) 3-{4-[(1H-Benzotriazole-5-carbonyl)amino]phenyl}acrylic acid 0 OH .' N N I& H H 10 HPLC-MS (Method A): m/z: 309 (M+1); Rt = 3.19 min. Example 44 (General procedure (A)) {3-[(1 H-Benzotriazole-5-carbonyl)amino]phenyl}acetic acid N' N OH H H N H 15 HPLC-MS (Method A): m/z: 297 (M+1); Rt = 2.10 min. Example 45 (General procedure (A)) 2-({4-[(1 H-Benzotriazole-5-carbonyl)amino]phenoxy}-2-methylpropionic acid 0 N" I N H 3 C CH 3 0 H H 20 HPLC-MS (Method A): m/z: 341 (M+1); Rt = 2.42 min. Example 46 (General procedure (A)) 3-{4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoylamino}propionic acid WO 2004/080480 PCT/DK2004/000158 117 0 0 r -. J N'-. OH N NNOH N I IH .N Ne H HPLC-MS (Method A): m/z: 354 (M+1); Rt = 1.78 min. Example 47 (General procedure (A)) 5 3-{4-[(1H-Benzotriazole-5-carbonyl)amino]phenyl}propionic acid
N
OH NN I, H N & H HPLC-MS (Method A): m/z: 311 (M+1); Rt = 2.20 min. Example 48 (General procedure (A)) 10 1H-Benzotriazole-5-carboxylic acid (4-benzyloxyphenyl)amide N N H HPLC-MS (Method A): m/z: 345 (M+1); Rt = 3.60 min. Example 49 (General procedure (A)) 15 1H-Benzotriazole-5-carboxylic acid (3-chloro-4-methoxyphenyl)amide N0 Cl'CH3 N I H c *N H H HPLC-MS (Method A): m/z: 303 (M+1); Rt = 2.88 min. Example 50 (General procedure (A)) 20 1H-Benzotriazole-5-carboxylic acid (4-phenoxyphenyl)amide WO 2004/080480 PCT/DK2004/000158 118 N* N N~I HH *N H HPLC-MS (Method A): m/z: 331 (M+1); Rt = 3.62 min. Example 51 (General procedure (A)) 5 1 H-Benzotriazole-5-carboxylic acid (4-butoxyphenyl)amide O ra' " O ' c H3 N , H C-I N lf H HPLC-MS (Method A): m/z: 311 (M+1); Rt = 3.59 min. Example 52 (General procedure (A)) 10 1H-Benzotriazole-5-carboxylic acid (3-bromo-4-trifluoromethoxyphenyl)amide O N Br F N' I a H N e H HPLC-MS (Method A): m/z: 402 (M+1); Rt = 3.93 min. Example 53 (General procedure (A)) 15 1 H-Benzotriazole-5-carboxylic acid (3,5-dichloro-4-hydroxyphenyl)amide Cl 0 OH NJ:N Cl N' N Ne N' H HPLC-MS (Method A): m/z: 323 (M+1); Rt = 2.57 min. Example 54 (General procedure (A)) 20 4-{[(1H-Benzotriazole-5-carbonyl)amino]methyl}benzoic acid WO 2004/080480 PCT/DK2004/000158 119 0 N N NOH HO HPLC-MS (Method A): m/z: 297 (M+1); Rt = 1.86 min. Example 55 (General procedure (A)) 5 {4-[(1H-Benzotriazole-5-carbonyl)amino]phenylsulfanyl}acetic acid SOH *N NI H N H HPLC-MS (Method A): m/z: 329 (M+1); Rt = 2.34 min. Example 56 10 N-(1H-Benzotriazol-5-yl)acetamide H N N ''CH3 H HPLC-MS (Method A): m/z: 177 (M+1); Rt = 0.84 min. Example 57 (General Procedure (A)) 15 1H-Benzotriazole-5-carboxylic acid 4-nitrobenzylamide H N N_ 0 The following compound is prepared according to general procedure (N) as described below: Example 58 (General procedure (N)) 20 1H-Benzotriazole-5-carboxylic acid 4-chlorobenzylamide 0 'N N ' C NI H H, Cl H HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.40 min.
WO 2004/080480 PCT/DK2004/000158 120 Example 59 2-[(1H-Benzotriazol-5-ylimino)methyl]-4,6-dichlorophenol H N ND N S OH Cl 5 Example 60 Diethyl 2-[(1H-benzotriazol-6-ylamino)methylidene]malonate
H
3 C) H H NN O N 0
CH
3 Example61 Nl-(1H-Benzotriazol-5-yl)-3-chlorobenzamide HN ~ 0 N.. HN N H CI 10 Example62 Nl-(1H-Benzotriazol-5-yl)-3,4,5-trimethoxybenzamide H
CH
3 0 N O1N N 0 N (:- -N H3Co H H3C '0 WO 2004/080480 PCT/DK2004/000158 121 Example 63 N2-(1H-Benzotriazol-5-yl)-3-chlorobenzo[b]thiophene-2-carboxamide 0 'N N °I H
S~
j' N NN C1 Example 64 6-Bromo-1 H-benzotriazole N Br N 5 H 5 Example 65 2-[(1 H-Benzotriazol-5-ylimino)methyl]-4-bromophenol H N NN Br OH 10 General procedure (B) for preparation of compounds of general formula 12: x X HN+ O A H HN A ' HN) + J H 0 0 R 3 12 wherein X, Y, A and R 3 are as defined above and A is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 as defined above. 15 The chemistry is well known (eg Lohray et al., J. Med. Chem., 1999, 42, 2569-81) and is generally performed by reacting a carbonyl compound (aldehyde or ketone) with the hetero cyclic ring (eg thiazolidine-2,4-dione (X = O; Y = S), rhodanine (X = Y = S) and hydantoin (X WO 2004/080480 PCT/DK2004/000158 122 = O; Y = NH) in the presence of a base, such as sodium acetate, potassium acetate, ammo nium acetate, piperidinium benzoate or an amine (eg piperidine, triethylamine and the like) in a solvent (eg acetic acid, ethanol, methanol, DMSO, DMF, NMP, toluene, benzene) or in a mixture of two or more of these solvents. The reaction is performed at room temperature or 5 at elevated temperature, most often at or near the boiling point of the mixture. Optionally, azeotropic removal of the formed water can be done. This general procedure (B) is further illustrated in the following example: Example 66 (General procedure (B)) 10 5-(3-Phenoxybenzylidene)thiazolidine-2,4-dione 0 HO o 00 A solution of thiazolidine-2,4-dione (90%, 78 mg, 0.6 mmol) and ammonium acetate (92 mg, 1.2 mmol) in acetic acid (1 mL) was added to 3-phenoxybenzaldehyde (52 pL, 0.6 mmol) and the resulting mixture was shaken at 115 OC for 16 hours. After cooling, the mixture was con 15 centrated in vacuo to afford the title compound. HPLC-MS (Method A): m/z: 298 (M+1); Rt = 4.54 min. The compounds in the following examples were similarly prepared. Optionally, the com 20 pounds can be further purified by filtration and washing with water, ethanol and / or heptane instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or heptane, or by chromatography, such as preparative HPLC. Example 67 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)thiazolidine-2,4-dione O CH HN N CH3 25 0 HPLC-MS (Method C): m/z: 249 (M+1); Rt = 4.90 min Example 68 (General procedure (B)) 5-Naphthalen-1 -ylmethylenethiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 123 HN o I HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,16 min. Example 69 (General procedure (B)) 5 5-Benzylidene-thiazolidine-2,4-dione 0 HN O HPLC-MS (Method A): m/z: 206 (M+1); Rt = 4,87 min. Example 70 (General procedure (B)) 10 5-(4-Diethylaminobenzylidene)thiazolidine-2,4-dione O
CH
3 o K HN N
CH
3 HN 0 HPLC-MS (Method A): m/z: 277 (M+1); Rt = 4.73 min. Example 71 (General procedure (B)) 15 5-(4-Methoxy-benzylidene)-thiazolidine-2,4-dione HN
CH
3 0 HPLC-MS (Method A): m/z: 263 (M+1); Rt = 4,90 min. Example 72 (General procedure (B)) 20 5-(4-Chloro-benzylidene)-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 124 0 HN Cl O HPLC-MS (Method A): m/z: 240 (M+1); Rt = 5,53 min. Example 73 (General procedure (B)) 5 5-(4-Nitro-benzylidene)-thiazolidine-2,4-dione O + 0 0+ HNO 0 HPLC-MS (Method A): m/z: 251 (M+1); Rt = 4,87 min. Example 74 (General procedure (B)) 10 5-(4-Hydroxy-3-methoxy-benzylidene)-thiazolidine-2,4-dione HN OH O 0 HPLC-MS (Method A): m/z: 252 (M+1); Rt = 4,07 min. Example 75 (General procedure (B)) 15 5-(4-Methylsulfanylbenzylidene)thiazolidine-2,4-dione O HN SCH3 0 HPLC-MS (Method A): m/z: 252 (M+1); Rt = 5,43 min. Example 76 (General procedure (B)) 20 5-(2-Pentyloxybenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 125
CH
3 O HNN 0 HPLC-MS (Method C): m/z: 292 (M+1); Rt = 4.75 min. 1 H NMR (DMSO-d 6 ): 6 = 0.90 (3H, t), 1.39 (4H, m), 1.77 (2H, p), 4.08 (2H, t), 7.08 (1H, t), 7.14 (1H, d), 7.43 (2H, m), 8.03 (1H, s), 12.6 (1H, bs). 5 Example 77 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)thiazolidine-2,4-dione HN O'CH O F 0 HPLC-MS (Method A): m/z: 354 (M+1); Rt = 4,97 min. 10 Example 78 (General procedure (B)) 5-(4-tert-Butylbenzylidene)thiazolidine-2,4-dione
H
3 C
CH
3 HN
CH
3 0 HPLC-MS (Method A): m/z: 262 (M+1); Rt = 6,70 min. 15 Example 79 (General procedure (B)) N-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acetamide 0 H H N CH 3 HNO - 0 0 HPLC-MS (Method A): m/z: 263 (M+1); Rt = 3,90 min. 20 WO 2004/080480 PCT/DK2004/000158 126 Example 80 (General procedure (B)) 5-Biphenyl-4-ylmethylene-thiazolidine-2,4-dione 0 oF HN 0 HPLC-MS (Method A): m/z: 282 (M+1); Rt = 4,52 min. 5 Example 81 (General procedure (B)) 5-(4-Phenoxy-benzylidene)-thiazolidine-2,4-dione 0 HN 0 HPLC-MS (Method A): m/z: 298 (M+1); Rt = 6,50 min. 10 Example 82 (General procedure (B)) 5-(3-Benzyloxybenzylidene)thiazolidine-2,4-dione 0 HNN 00 HPLC-MS (Method A): m/z: 312 (M+1); Rt = 6,37 min. 15 Example 83 (General procedure (B)) 5-(3-p-Tolyloxybenzylidene)thiazolidine-2,4-dione o N z
CH
3 HPL-MS (Method A): m/z: 312 (M); Rt = 6,87 in. 20 HPLC-MS (Method A): mlz: 312 (M+1); Rt =6,87 min. 20 Example 84 (General procedure (B)) 5-Naphthalen-2-ylmethylene-thiazolidine-2,4-dione 0 HN 0 WO 2004/080480 PCT/DK2004/000158 127 HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4.15 min. Example 85 (General procedure (B)) 5-Benzo[1,3]dioxol-5-ylmethylenethiazolidine-2,4-dione 0 O HN > 50 5 0 HPLC-MS (Method A): m/z: 250 (M+1), Rt = 3.18 min. Example 86 (General procedure (B)) 5-(4-Chlorobenzylidene)-2-thioxothiazolidin-4-one H-s C l HN 10 O HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,51 min. Example 87 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)-2-thioxothiazolidin-4-one CHz H N' CH3 15 0 HPLC-MS (Method A): m/z: 265 (M+1); Rt = 5,66 min. Example 88 (General procedure (B)) 5-(4-Nitrobenzylidene)-2-thioxothiazolidin-4-one O HN 20 0 HPLC-MS (Method A): m/z: 267 (M+1); Rt = 3,94 min.
WO 2004/080480 PCT/DK2004/000158 128 Example 89 (General procedure (B)) 5-(4-Methylsulfanylbenzylidene)-2-thioxothiazolidin-4-one HNs S'CH3 0 HPLC-MS (Method A): m/z: 268 (M+1); Rt = 6,39 min. 5 Example 90 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)-2-thioxothiazolidin-4-one s H N ' c H 3 N K 0 HPLC-MS (Method A): m/z: 270 (M+1); Rt = 5,52 min. 10 Example 91 (General procedure (B)) 5-Naphthalen-2-ylmethylene-2-thioxothiazolidin-4-one S O 0 HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,75 min. 15 Example 92 (General procedure (B)) 5-(4-Diethylaminobenzylidene)-2-thioxothiazolidin-4-one r
CH
3 H N CH3 HN ' NcH 0 HPLC-MS (Method A): m/z: 293 (M+1); Rt = 5,99 min. 20 Example 93 (General procedure (B)) 5-Biphenyl-4-ylmethylene-2-thioxothiazolidin-4-one WO 2004/080480 PCT/DK2004/000158 129 Ss HN O HPLC-MS (Method A): m/z: 298 (M+1); Rt = 7,03 min. Example 94 (General procedure (B)) 5 5-(3-Phenoxybenzylidene)-2-thioxothiazolidin-4-one S HN O O HPLC-MS (Method A): m/z: 314 (M+I); Rt = 6,89 min. Example 95 (General procedure (B)) 10 5-(3-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one o HPLC-MS (Method A): m/z: 328 (M+1); Rt = 6,95 min. Example 96 (General procedure (B)) 15 5-(4-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one s HN NIa 0 HPLC-MS (Method A): m/z: 328 (M+1); RT = 6,89 min. Example 97 (General procedure (B)) 20 5-Naphthalen-1 -ylmethylene-2-thioxothiazolidin-4-one S HN 0H HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,43 min.
WO 2004/080480 PCT/DK2004/000158 130 Example 98 (General procedure (B)) 5-(3-Methoxybenzyl)thiazolidine-2,4-dione 0 HN 0oCH 3 O 5 HPLC-MS (Method A): m/z: 236 (M+1); Rt = 3,05 min. Example 99 (General procedure (D)) 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid ethyl ester O 10 HPLC-MS (Method A): m/z: 392 (M+23), Rt = 4.32 min. Example 100 (General procedure (D)) 4 -[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phenoxy]-butyric acid 0 HN I # v v Br 0 HOO 15 HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3,35 min. Example 101 (General procedure (B)) 5-(3-Bromobenzylidene)thiazolidine-2,4-dione H Br O 0 20 HPLC-MS (Method A): mlz: 285 (M+1); Rt = 4.01 min. Example 102 (General procedure (B)) 5-(4-Bromobenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 131 0 HN Br O HPLC-MS (Method A): m/z: 285 (M+1); Rt = 4.05 min. Example 103 (General procedure (B)) 5 5-(3-Chlorobenzylidene)thiazolidine-2,4-dione 0 HN C O HPLC-MS (Method A): m/z: 240 (M+1); Rt = 3.91 min. Example 104 (General procedure (B)) 10 5-Thiophen-2-ylmethylenethiazolidine-2,4-dione 0 HNO O 0 HPLC-MS (Method A): m/z: 212 (M+1); Rt = 3.09 min. Example 105 (General procedure (B)) 15 5-(4-Bromothiophen-2-ylmethylene)thiazolidine-2,4-dione O S Br HN S 0 HPLC-MS (Method A): m/z: 291 (M+1); Rt = 3.85 min. Example 106 (General procedure (B)) 20 5-(3,5-Dichlorobenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 132 O- CI Cl O HPLC-MS (Method A): m/z: 274 (M+1); Rt = 4.52 min. Example 107 (General procedure (B)) 5 5-(1-Methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione 0 PH 3 HN 0O HPLC-MS (Method A): m/z: 259 (M+1); Rt = 3.55 min. Example 108 (General procedure (B)) 10 5-(1 H-Indol-3-ylmethylene)thiazolidine-2,4-dione HN NH O 0 HPLC-MS (Method A): m/z: 245 (M+1); Rt = 2.73 min. Example 109 (General procedure (B)) 15 5-Fluoren-9-ylidenethiazolidine-2,4-dione O HN 0 HPLC-MS (Method A): mlz: 280 (M+1); Rt =4.34 min.
WO 2004/080480 PCT/DK2004/000158 133 Example 110 (General procedure (B)) 5-(1-Phenylethylidene)thiazolidine-2,4-dione O YS HN O CH 3 HPLC-MS (Method A): m/z: 220 (M+1); Rt = 3,38 min. 5 Example 111 (General procedure (B)) 5-[1 -(4-Methoxyphenyl)-ethylidene]-thiazolidine-2,4-dione
CH
3 HN O CH 3 HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3.55 min. 10 Example 112 (General procedure (B)) 5-(1-Naphthalen-2-yl-ethylidene)-thiazolidine-2,4-dione O HN O CH 3 HPLC-MS (Method A): m/z: 270 (M+1); Rt = 4,30 min. 15 Example 113 (General procedure (B)) 5-[1-(4-Bromophenyl)-ethylidene]-thiazolidine-2,4-dione X~S'NBr HN O
CH
3 HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,18 min. 20 WO 2004/080480 PCT/DK2004/000158 134 Example 114 (General procedure (B)) 5-(2,2-Diphenylethylidene)-thiazolidine-2, 4 -dione 0 HN HPLC-MS (Method A): m/z: 296 (M+1); Rt = 4,49 min. 5 Example 115 (General procedure (B)) 5-[1-(3-Methoxyphenyl)-ethylidene]-thiazolidine-2,4-dione 0 HN 0oCH O CH, HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3,60 min. 10 Example 116 (General procedure (B)) 5-[1-(6-Methoxynaphthalen-2-yl)-ethylidene]-thiazolidine-2,4-dione o CH 3 - - 0 tiN O CH 3 HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,26 min. 15 Example 117 (General procedure (B)) 5-[1-(4-Phenoxyphenyl)-ethylidene]-thiazolidine-2,4-dione 0 O CH 3 HPLC-MS (Method A): m/z: 312 (M+1); Rt = 4,68 min. 20 Example 118 (General procedure (B)) 5-[1-(3-Fluoro-4-methoxyphenyl)ethylidene]thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 135 H '1CHz O CH 3 HPLC-MS (Method A): m/z: 268 (M+1); Rt = 3,58 min. Example 119 (General procedure (B)) 5 5-[1-(3-Bromophenyl)-ethylidene]-thiazolidine-2,4-dione 0 HN H a Br O
CH
3 HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,13 min. Example 120 (General procedure (B)) 10 5-Anthracen-9-ylmethylenethiazolidine-2,4-dione HN 0 I o HPLC-MS (Method A): m/z: 306 (M+1); Rt = 4,64 min. Example 121 (General procedure (B)) 15 5-(2-Methoxynaphthalen-1 -ylmethylene)-thiazolidine-2,4-dione
CH
3 OS HN 0 HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4,02 min. Example 122 (General procedure (B)) 20 5-(4-Methoxynaphthalen-1 -ylmethylene)-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 136 0 HNs '0CH 3 HN ' O HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4,31 min. Example 123 (General procedure (B)) 5 5-(4-Dimethylaminonaphthalen-1 -ylmethylene)-thiazolidine-2,4-dione O CH 3 HN S N'CH 3 HN 0 HPLC-MS (Method A): m/z: 299 (M+1); Rt = 4,22 min. Example 124 (General procedure (B)) 10 5-(4-Methylnaphthalen-1-ylmethylene)-thiazolidine-2,4-dione 0 HNS
CH
3 0/ HPLC-MS (Method A): m/z: 270 (M+1); Rt = 4,47 min. Example 125 (General procedure (B)) 15 5-Pyridin-2-ylmethylene-thiazolidine-2,4-dione 0 HN N O Example 126 5-Pyridin-2-ylmethyl-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 137 O S HN N 0 5-Pyridin-2-ylmethylene-thiazolidine-2,4-dione (5 g) in tetrahydrofuran (300 ml) was added 10% Pd/C (1 g) and the mixture was hydrogenated at ambient pressure for 16 hours. More 10% Pd/C (5 g) was added and the mixture was hydrogenated at 50 psi for 16 hours. After 5 filtration and evaporation in vacuo, the residue was purified by column chromatography eluting with a mixture of ethyl acetate and heptane (1:1). This afforded the title compound (0.8 g, 16%) as a solid. TLC: Rf = 0.30 (SiO 2 ; EtOAc: heptane 1:1) 10 Example 127 (General procedure (B)) 5-(1 H-Imidazol-4-ylmethylene)-thiazolidine-2,4-dione O-S N HN ,,NH O 0 15 Example 128 (General procedure (B)) 5-(4-Benzyloxy-benzylidene)-thiazolidine-2,4-dione HN o 0 HPLC-MS (Method A): m/z: 6,43 min; 99 % (2A) 20 Example 129 (General procedure (B)) 5-[4-(4-Fluorobenzyloxy)benzylidene]-2-thioxothiazolidin-4-one OF 0 WO 2004/080480 PCT/DK2004/000158 138 Example 130 (General procedure (B)) 5-(4-Butoxybenzylidene)-2-thioxothiazolidin-4-one CH3
O
0 H 0 5 Example 131 (General procedure (B)) 5-(3-Methoxybenzylidene)thiazolidine-2,4-dione 0 HN;OCH3 O 10 HPLC-MS (Method A): m/z: 236 (M+1); Rt = 4,97 min Example 132 (General procedure (B)) 15 5-(3-Methoxybenzylidene)imidazolidine-2,4-dione -N oNOH HNI H O.CH3 HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.43 min. Example 133 (General procedure (B)) 20 5-(4-Methoxybenzylidene)imidazolidine-2,4-dione
O\
H - 0. HN O CH 3 HPLC-MS (Method A): m/z: 219 (M+0); Rt = 2.3 min. HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.38 min.
WO 2004/080480 PCT/DK2004/000158 139 Example 134 (General procedure (B)) 5-(2,3-Dichlorobenzylidene)thiazolidine-2,4-dione 0 HIN ClI O Cl 5 Example 135 (General procedure (B)) 5-Benzofuran-7-ylmethylenethiazolidine-2,4-dione 0 0
-
-~I HN -b 0 HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4,57 min. 10 Example 136 (General procedure (B)) 5-Benzo[1,3]dioxol-4-ylmethylenethiazolidine-2,4-dione o-O 0 rO HN 0 HPLC-MS (Method C): m/z: 250 (M+1); Rt = 4,00 min. 15 Example 137 (General procedure (B)) 5-(4-Methoxy-2,3-dimethylbenzylidene)thiazolid ine-2,4-dione 0
CH
3 H O HN CH 3 O CH 3 HPLC-MS (Method C): m/z: 264 (M+1); Rt = 5,05 min. 20 Example 138 (General procedure (B)) 5-( 2 -Benzyloxy-3-methoxybenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 140 0 HN H3JaH 06 HPLC-MS (Method C): m/z: 342 (M+1); Rt = 5,14 min. Example 139 (General procedure (B)) 5 5-(2-Hydroxybenzylidene)thiazolidine-2,4-dione O HN O OH HPLC-MS (Method C): m/z: 222 (M+I1); Rt = 3,67 min. Example 140 (General procedure (B)) 10 5-(2,4-Dichlorobenzylidene)thiazolidine-2,4-dione O Cl HN O Cl 'H-NMR (DMSO-d 6 ): 7.60 (2H, "s"), 7.78 (1H, s), 7.82 (1H, s). Example 141 (General procedure (B)) 15 5-(2-Chlorobenzylidene)thiazolidine-2,4-dione H O CI 1 H-NMR (DMSO-d 6 ): 7.40 (1H, t), 7.46 (1H, t), 7.57 (1H, d), 7.62 (1H, d), 7.74 (1H, s). Example 142 (General procedure (B)) 20 5-(2-Bromobenzylidene)thiazolidine-2,4-dione 0 HN O Br 'H-NMR (DMSO-d6): 7.33 (1H, t), 7.52 (1H, t), 7.60 (1H, d), 7.71 (1H, s), 7.77 (1H, d).
WO 2004/080480 PCT/DK2004/000158 141 Example 143 (General procedure (B)) 5-(2,4-Dimethoxybenzylidene)thiazolidine-2,4-dione CH3 O 0
HNL
O O'CH3 5 HPLC-MS (Method C): m/z: 266 (M+1) Rt = 4,40 min. Example 144 (General procedure (B)) 5-(2-Methoxybenzylidene)thiazolidine-2,4-dione 0 -s " H N 0 O'CH3 10 HPLC-MS (Method C): m/z: 236 (M+1); Rt = 4,17 min. Example 145 (General procedure (B)) 5-(2,6-Difluorobenzylidene)thiazolidine-2,4-dione HN o F 15 HPLC-MS (Method C): m/z: 242 (M+1); Rt = 4,30 min. Example 146 (General procedure (B)) 5-(2,4-Dimethylbenzylidene)thiazolidine-2,4-dione 0 O H 3 HN O H 0 CH3 20 HPLC-MS (Method C): m/z: 234 (M+1); Rt = 5,00 min. Example 147 (General procedure (B)) 5-(2,4,6-Trimethoxybenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 142 OH CH, o s 0 0 HN O O
O'CH
3 HPLC-MS (Method C): m/z: 296 (M+1); Rt = 4,27 min. Example 148 (General procedure (B)) 5 5-(4-Hydroxy-2-methoxybenzylidene)thiazolidine-2,4-dione O OH 0
O-CH
3 HPLC-MS (Method C): m/z: 252 (M+1); Rt = 3,64 min. Example 149 (General procedure (B)) 10 5-(4-Hydroxynaphthalen-1-ylmethylene)thiazolidine-2,4-dione O OH HN 0 'H-NMR (DMSO-d): 6 = 7.04 (1H, d), 7.57 (2H, m), 7.67 (1H, t), 8.11 (1H, d), 8.25 (1H, d), 8.39 (1H, s) 11.1 (1H, s), 12.5 (1H, bs). HPLC-MS (Method C): m/z: 272 (M+1); Rt = 3.44 mm. 15 Example 150 (General procedure (B)) 5-(2-Trifluoromethoxybenzylidene)thiazolidine-2,4-dione HN -s ' ,I O HN- F F HPLC-MS (Method C): mlz: 290 (M+1); Rt = 4,94 min. 20 Example 151 (General procedure (B)) 5-Biphenyl-2-ylmethylenethiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 143 O HN s O HPLC-MS (Method C): m/z: 282 (M+1); Rt = 5,17 min. Example 152 (General procedure (B)) 5 5-(2-Benzyloxybenzylidene)thiazolidine-2,4-dione 0 HNrj l~. H NO oH0 HPLC-MS (Method C): m/z: 312 (M+1); Rt = 5,40 min. Example 153 (General procedure (B)) 10 5-Adamantan-2-ylidenethiazolidine-2,4-dione O HN HPLC-MS (Method A): m/z: 250 (M+1); Rt = 4,30 min. Example 154 (General Procedure (B)) 15 5-[3-(4-Nitrophenyl)allylidene]thiazolidine-2,4-dione 0 %I 0 NO O N HN ' 0 HPLC-MS (Method C): m/z: 277 (M+1); Rt = 3.63 min. Example 155 (General Procedure (B)) 20 5-[3-(2-Methoxyphenyl)allylidene]thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 144
CH
3 HN s 0
'
0 HPLC-MS (Method C): m/z: 262 (M+1); Rt = 3.81 min. Example 156 (General Procedure (B)) 5 5-[3-(4-Methoxyphenyl)allylidene]thiazolidine-2,4-dione CH3 0 ~0 HN O 0 HPLC-MS (Method C): m/z: 262 (M+1); Rt = 3.67 min. Example 157 (General procedure (B)) 10 5-(4-Hydroxybenzylidene)thiazolidine-2,4-dione HO H N S Example 158 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)pyrimidine-2,4,6-trione 55 1NH N0 N 0 H 15 CH 3 HPLC-MS (Method C): m/z = 260 (M+1) Rt = 2,16 min. Example 159 (General procedure (B)) 5-(9-Ethyl-9H-carbazol-2-ylmethylene)-pyrimidine-2,4,6-trione WO 2004/080480 PCT/DK2004/000158 145 H 0oy No 0 HN \ -\ N
SH
3 C HPLC-MS (Method C): m/z = 334 (M+I1); Rt = 3,55 min. Example 160 (General procedure (B)) 5 5-(4-Hexyloxynaphthalen-1 -ylmethylene)thiazolidine-2,4-dione O 1 0 ,
CH
3 H N 0 HPLC-MS (Method C): m/z = 356 (M+1); Rt = 5.75 min. Example 161 (General procedure (B)) 10 5-(4-Decyloxynaphthalen-1 -ylmethylene)thiazolidine-2,4-dione O HNN 0 HPLC-MS (Method C): m/z = 412 (M+1); Rt = 6.44 min. Example 162 (General procedure (B)) 15 5-[4-(2-Aminoethoxy)-naphthalen-1 -ylmethylene]-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 146 o HN S I
NH
2 0 HPLC-MS (Method C): m/z = 315 (M+1); Rt = 3,24 min. Example 163 (General procedure (B)) 5 5-(2,4-Dimethyl-9H-carbazol-3-ylmethylene)-pyrimidine-2,4,6-trione
H
3 C H 0 N 0 NH HN \~ O CH 3 HPLC-MS (Method C): m/z = 334 (M+1); Rt = 3,14 min. Example 164 (General procedure (B)) 10 4-(4-Hydroxy-3-methoxybenzylidine)hydantoin 0 OO NH HO H O Example 165 (General procedure (B)) 5-Benzylidenehydantoin 0 NH 15 O General procedure (C) for preparation of compounds of general formula 12: WO 2004/080480 PCT/DK2004/000158 147 X 0 A, x_ _y O' A H HN HN + ' H HN AH R O R 0 12 wherein X, Y, A, and R 3 are as defined above and A is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 as defined above. 5 This general procedure (C) is quite similar to general procedure (B) and is further illustrated in the following example: Example 166 (General procedure (C)) 5-(3,4-Dibromobenzylidene)thiazolidine-2,4-dione O~s T 'T B r 0 HN Br 10 0 A mixture of thiazolidine-2,4-dione (90%, 65 mg, 0.5 mmol), 3,4-dibromobenzaldehyde (132 mg, 0.5 mmol), and piperidine (247 pL, 2.5 mmol) was shaken in acetic acid (2 mL) at 11 0 OC for 16 hours. After cooling, the mixture was concentrated to dryness in vacuo. The resulting crude product was shaken with water, centrifuged, and the supernatant was 15 discarded. Subsequently the residue was shaken with ethanol, centrifuged, the supernatant was discarded and the residue was further evaporated to dryness to afford the title com pound. 1 H NMR (Acetone-d 6 ): 6H 7.99 (d,1H), 7.90 (d,1H), 7.70 (s,1H), 7.54 (d,1H); HPLC-MS 20 (Method A): m/z: 364 (M+1); Rt = 4.31 min. The compounds in the following examples were similarly prepared. Optionally, the com pounds can be further purified by filtration and washing with water instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or 25 heptane, or by preparative HPLC. Example 167 (General procedure (C)) 5-(4-Hydroxy-3-iodo-5-methoxybenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 148 HNO OH N . o-oCH 3 O Mp = 256 oC; 1 H NMR (DMSO-d) 6 = 12.5 (s,broad,1H), 10.5 (s,broad,1H), 7.69 (s,IH), 7.51 (d,1H), 7.19 (d,1H)3.88 (s,3H), 1 3 C NMR (DMSO-d 6 ) 6c = 168.0, 167.7,149.0, 147.4, 133.0, 131.2, 126.7, 121.2, 113.5, 85.5, 56.5; HPLC-MS (Method A): m/z: 378 (M+1); Rt = 3.21 min. 5 Example 168 (General procedure (C)) 5-(4-Hydroxy-2,6-dimethylbenzylidene)thiazolidine-2,4-dione O _S 3"H~C OH HN OH O
CH
3 10 HPLC-MS (Method C): m/z: 250 (M+1); Rt.= 2.45 min. Example 169 (General procedure (C)) 4-[5-Bromo-6-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-2-yloxymethyl]-benzoic acid OH o oo O Br 15 HPLC-MS (Method C): m/z: 506 (M+23); Rt.= 4.27 min. Example 170 (General procedure (C)) 5-(4-Bromo-2,6-dichlorobenzylidene)thiazolidine-2,4-dione H C ! Br O C B HN N 20 0 Cl HPLC-MS (Method C): m/z: 354 (M+1); Rt.= 4.36 min.
WO 2004/080480 PCT/DK2004/000158 149 Example 171 (General procedure (C)) 5-(6-Hydroxy-2-naphthylmethylene) thiazolidine-2,4-dione 0 H OH HN X 0 Mp 310-314 oC, 'H NMR (DMSO-d): 6 H = 12.5 (s,broad,1H), 8.06(d,1H), 7.90 5 7.78(m,2H),7.86 (s, 1H), 7.58 (dd,1 H),7.20 7.12 (m,2H). 1 3 C NMR (DMSO-d): 6c = 166.2, 165.8, 155.4, 133.3, 130.1, 129.1, 128.6, 125.4, 125.3, 125.1,124.3, 120.0, 117.8, 106.8; HPLC-MS (Method A): m/z: 272 (M+1); Rt = 3.12 min. Preparation of the starting material, 6-hydroxy-2-naphtalenecarbaldehyde: 10 6-Cyano-2-naphthalenecarbaldehyde (1.0 g, 5.9 mmol) was dissolved in dry hexane (15 mL) under nitrogen. The solution was cooled to -60 oC and a solution of diisobutyl aluminium hy dride (DIBAH) (15 mL, 1M in hexane) was added dropwise. After the addition, the solution was left at room temperature overnight. Saturated ammonium chloride solution (20 mL) was 15 added and the mixture was stirred at room temperature for 20 min, subsequently aqueous
H
2
SO
4 (10% solution, 15 mL) was added followed by water until all salt was dissolved. The resulting solution was extracted with ethyl acetate (3x), the combined organic phases were dried with MgSO 4 , evaporated to dryness to afford 0.89 g of 6-hydroxy-2 naphtalenecarbaldehyde. 20 Mp.: 153.5-156.5 Oc; HPLC-MS (Method A): m/z: 173 (M+1); Rt = 2.67 min; ' H NMR (DMSO d 6 ): 6 H = 10.32(s,1H), 8.95 (d,1H), 10.02 (s,1H), 8.42 (s,broad,1H), 8.01 (d,1H), 7.82-7.78 (m,2H), 7.23-7.18 (m,2H). 25 Alternative preparation of 6-hydroxy-2-naphtalenecarbaldehyde: To a stirred cooled mixture of 6-bromo-2-hydroxynaphthalene (25.3 g, 0.113 mol) in THF (600 mL) at -78 OC was added n-BuLi (2.5 M, 100 mL, 0.250 mol) dropwise. The mixture turned yellow and the temperature rose to -64 OC. After ca 5 min a suspension appeared. After addition, the mixture was maintained at -78 oC. After 20 minutes, a solution of DMF 30 (28.9 mL, 0.373 mol) in THF (100 mL) was added over 20 minutes. After addition, the mi; ture was allowed to warm slowly to room temperature. After 1 hour, the mixture was poured in ice/water (200 mL). To the mixture citric acid was added to a pH of 5. The mixture was stirred for 0.5 hour. Ethyl acetate (200 mL) was added and the organic layer was separated WO 2004/080480 PCT/DK2004/000158 150 and washed with brine (100 mL), dried over Na 2
SO
4 and concentrated. To the residue was added heptane with 20% ethyl acetate (ca 50 mL) and the mixture was stirred for 1 hour. The mixture was filtered and the solid was washed with ethyl acetate and dried in vacuo to afford 16 g of the title compound. 5 Example 172 (General procedure (C)) 5-(3-lodo-4-methoxybenzylidene)thiazolidiene-2,4-dione 0 0 CH3 HN 0 1 H NMR (DMSO-d 6 ): 6 H 12.55 (s,broad,1H), 8.02 (d,1H), 7.72 (s,1H), 7.61 (d,1H)7.18(d,1H), 10 3.88 (s,3H); 13 C NMR (DMSO-d): 6 c 168.1, 167.7, 159.8, 141.5, 132.0, 130.8, 128.0, 122.1, 112.5, 87.5, 57.3. HPLC-MS (Method A): m/z: 362 (M+1); Rt = 4.08 min. Preparation of the starting material, 3-iodo-4-methoxybenzaldehyde: 15 4-Methoxybenzaldehyde (0.5 g, 3.67 mmol) and silver trifluoroacetate (0.92 g, 4.19 mmol) were mixed in dichloromethane (25 mL). Iodine (1.19 g, 4.7 mmol) was added in small por tions and the mixture was stirred overnight at room temperature under nitrogen. The mixture was subsequently filtered and the residue washed with DCM. The combined filtrates were treated with an acqueous sodium thiosulfate solution (1 M) until the colour disappeared. 20 Subsequent extraction with dichloromethane (3 x 20 mL) followed by drying with MgSO 4 and evaporation in vacuo afforded 0.94 g of 3-iodo-4-methoxybenzaldehyde. Mp 104-107 °C; HPLC-MS (Method A): m/z:263 (M+1); Rt = 3.56 min.;'H NMR (CDCl 3 ): 6H = 8.80 (s,1H), 8.31 (d,lH), 7.85 (dd,lH) 6.92 (d,1H), 3.99 (s, 3H). 25 Example 173 (General procedure (C)) 5-(1-Bromonaphthalen-2-ylmethylene)thiazolidine-2,4-dione HN O Br HPLC-MS (Method A): m/z: =336 (M+1); Rt = 4.46 min. 30 WO 2004/080480 PCT/DK2004/000158 151 Example 174 (General procedure (C)) 1-[5-(2,4-Dioxothiazolidin-5-ylidenemethyl)thiazol-2-yl]piperidine-4-carboxylic acid ethyl ester H33 0o ON SHN O H 'H NMR (DMSO-d 6 ): 6 H = 7.88 (s,1H), 7.78 (s,1H), 4.10 (q,2H), 4.0-3.8 (m,2H), 3.40-3.18 5 (m,2H), 2.75-2.60 (m,1H), 2.04-1.88 (m,2H), 1.73-1.49 (m,2H), 1.08 (t,3H); HPLC-MS (Method A): m/z: 368 (M+1); Rt = 3.41 min. Example 175 (General procedure (C)) 5-(2-Phenyl-[1,2,3]triazol-4-ylmethylene) thiazolidine-2,4-dione 0 H N HN _N 'N 10 o 'H NMR (DMSO-d 6 ): 6 H = 12.6 (s,broad,lH), 8.46 (s,1H), 8.08 (dd,2H), 7.82 (s,1 H), 7.70-7.45 (m, 3H). HPLC-MS (Method A): m/z: 273 (M+1); Rt = 3.76 min. Example 176 (General procedure (C)) 15 5-(Quinolin-4-ylmethylene)thiazolidine-2,4-dione 0 O-S IN HN I 0 O HPLC-MS (Method A): m/z: 257 (M+1); Rt = 2.40 min. Example 177 (General procedure (C)) 20 5-(6-Methylpyridin-2-ylmethylene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 152 O
CH
3 SN HN o 1 H NMR (DMSO-d 6 ): 6 H = 12.35 (s,broad,1H), 7.82 (t,1H), 7.78 (s,1H), 7.65 (d,1H), 7.18 (d,1H), 2.52 (s,3 H); HPLC-MS (Method A): m/z: 221 (M+1); Rt = 3.03 min. 5 Example 178 (General procedure (C)) 5-(2,4-dioxothiazolidin-5-ylidenemethyl)-furan-2-ylmethylacetate 0 H O , o CH3 O N\f 0 0 0 1 H NMR (DMSO-d): bH = 12.46 (s,broad,1H), 7.58 (s,1H), 7.05 (d,1H), 6.74 (s,1 H), 5.13 (s,2H), 2.10 (s,3H). HPLC-MS (Method A): m/z: 208 (M-CHaCOO); Rt = 2.67 min. 10 Example 179 (General procedure (C)) 5-(2,4-Dioxothiazolidin-5-ylidenemethyl)furan-2-sulfonic acid o-s HN / 0 H o 0 HPLC-MS (Method A): m/z:276 (M+1); Rt = 0.98 min. 15 Example 180 (General procedure (C)) 5-(5-Benzyloxy-1 H-pyrrolo[2,3-c]pyridin-3-ylmethylene)-thiazolidine-2,4-dione HNo O0 HPLC-MS (Method A): m/z: 352 (M+1); Rt = 3.01 min. 20 Example 181 (General procedure (C)) 5-(Quinolin-2-ylmethylene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 153 O HNN 0 HPLC-MS (Method A): m/z: 257 (M+1); Rt = 3.40 min. Example 182 (General procedure (C)) 5 5-(2,4-Dioxothiazolidin-5-ylidenemethyl)thiophene-2-carboxylic acid 0 HN o O OH HPLC-MS (Method A): m/z: 256 (M+1); Rt = 1.96 min. Example 183 (General procedure (C)) 10 5-(2-Phenyl-1 H-imidazol-4-ylmethylene)thiazolidine-2,4-dione H H N HN!:,3 N O HPLC-MS (Method A): m/z: 272 (M+1); Rt = 2.89 min. Example 184 (General procedure (C)) 15 5-(4-Imidazol-1-yl-benzylidene)thiazolidine-2,4-dione s HN N O 0 HPLC-MS (Method A): m/z: 272 (M+1); Rt = 1.38 min. Example 185 (General procedure (C)) 20 5-(9-Ethyl-9H-carbazol-3-ylmethylene)thiazolidine-2,4-dione rCH 3 HN N 0 WO 2004/080480 PCT/DK2004/000158 154 HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.52 min. Example 186 (General procedure (C)) 5-(1,4-Dimethyl-9H-carbazol-3-ylmethylene)thiazolidine-2,4-dione 0 H 3 C H o 5 0 CH 3 HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.35 min. Example 187 (General procedure (C)) 5-(2-Methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione O HC H HN 10 O HPLC-MS (Method A): m/z: 259 (M+1); Rt = 3.24 min. Example 188 (General procedure (C)) 5-(2-Ethylindol-3-ylmethylene)thiazolidine-2,4-dione
OCH
3 HN 15 o 2-Methylindole (1.0 g, 7.6mmol) dissolved in diethyl ether (100 mL) under nitrogen was treated with n-Butyl lithium (2 M in pentane, 22.8 mmol) and potassium tert-butoxide (15.2 mmol) with stirring at RT for 30 min. The temperature was lowered to -70 C and methyl lo 20 dide (15.2 mmol) was added and the resulting mixture was stirred at -70 for 2 h. Then 5 drops of water was added and the mixture allowed to warm up to RT. Subsequently, the mix ture was poured into water (300 mL), pH was adjusted to 6 by means of 1N hydrochloric acid and the mixture was extracted with diethyl ether. The organic phase was dried with Na 2
SO
4 and evaporated to dryness. The residue was purified by column chromatography on silica gel 25 using heptane/ether(4/1) as eluent. This afforded 720 mg (69 %) of 2-ethylindole.
WO 2004/080480 PCT/DK2004/000158 155 1 H NMR (DMSO-d 6 ): 8 = 10.85 (1H,s); 7.39 (1H,d); 7.25 (1H,d); 6.98(1H,t); 6.90(1H,t); 6.10 (1H,s); 2.71 (2H,q); 1.28 (3H,t). 2-Ethylindole (0.5 g, 3.4mmol) dissolved in DMF (2 mL) was added to a cold (0 OC) premixed 5 (30 minutes) mixture of DMF (1.15 mL) and phosphorous oxychloride (0.64 g, 4.16 mmol). After addition of 2-ethylindole, the mixture was heated to 40 OC for 1 h, water (5 mL) was added and the pH adjusted to 5 by means of 1 N sodium hydroxide.The mixture was subse quently extracted with diethyl ether, the organic phase isolated, dried with MgSO 4 and evapo rated to dryness affording 2-ethylindole-3-carbaldehyde (300 mg). 10 HPLC-MS (Method C): m/z:174 (M+1); Rt. =2.47 min. 2-Ethylindole-3-carbaldehyde (170 mg) was treated with thiazolidine-2,4-dione using the general procedure (C) to afford the title compound (50 mg). 15 HPLC-MS (Method C):m/z: 273 (M+1); Rt.= 3.26 min. Example 189 (General procedure (C)) 5-[2-(4-Bromophenylsulfanyl)-1-methyl-1 H-indol-3-ylmethylene]thiazolidine-2,4-dione Br C H3 HN 20 O HPLC-MS (Method A): m/z: 447 (M+1); Rt = 5.25 min. Example 190 (General procedure (C)) 5-[2-(2,4-Dichlorobenzyloxy)-naphthalen-1 -ylmethylene]thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 156 CI O /CI 0 o HN /\ o HPLC-MS (Method A): (anyone 1) m/z: 430 (M+1); Rt = 5.47 min. Example 191 (General procedure (C)) 5 5-{4-[3-(4-Bromophenyl)-3-oxopropenyl]-benzylidene}thiazolidine-2,4-dione 0 oF s HN Br 0 O HPLC-MS (Method A): m/z: 416 (M+1); Rt = 5.02 min. Example 192 (General procedure (C)) 10 5-(4-Pyridin-2-ylbenzylidene)thiazolidine-2,4-dione o HN N 0 HPLC-MS (Method A): m/z: 283 (M+1), Rt = 2.97 min. Example 193 (General procedure (C)) 15 5-(3,4-Bisbenzyloxybenzylidene)thiazolidine-2,4-dione 0 HN 0 HPLC-MS (Method A): m/z: 418 (M+1); Rt = 5.13 min. Example 194 (General procedure (C)) 20 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 157 0 II, 00 HN o HPLC-MS (Method A): m/z: 357 (M+1); Rt = 4.45 min. Example 195 (General procedure (C)) 5 5-(2-Phenyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione o /~ HNN HN I o HPLC-MS (Method A): m/z: 321 (M+1); Rt = 3.93 min. Example 196 (General procedure (C)) 10 5-(5-Benzyloxy-1H-indol-3-ylmethylene)thiazolidine-2,4-dione 0 H 00 ob HPLC-MS (Method A): m/z: 351 (M+1); Rt = 4.18 min. Example 197 (General procedure (C)) 15 5-(4-Hydroxybenzylidene)thiazolidine-2,4-dione HN OH O 0 HPLC-MS (Method A): m/z: 222 (M+1); Rt = 2.42 min. Example 198 (General procedure (C)) 5-(1-Methyl-lH-indol-2-ylmethylene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 158 0 N o CH 3 1 H NMR (DMSO-d 6 ): 6 H = 12.60 (s,broad,1H), 7.85 (s,1H), 7.68 (dd,lH), 7.55 (dd,1H), 7.38 (dt, 1H), 7.11 (dt, 1H) 6.84 (s, 1H), 3.88 (s,3H); HPLC-MS (Method A): m/z: 259 (M+1); Rt = 4.00 min. 5 Example 199 (General procedure (C)) 5-(5-Nitro-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione O H H N N HN + oNo C O .Mp 330-333 oC, 'H NMR (DMSO-d 6 ): 6H = 12.62 (s,broad,1lH), 8.95 (d,1IH), 8.20 (s,IH), 8.12 10 (dd,IH), 7.98 (s,broad,1H), 7.68 (d,1H); HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.18 min. Example 200 (General procedure (C)) 5-(6-Methoxynaphthalen-2-ylmethylene)thiazolidine-2,4-dione O H 3C HNN
.
-. 15 O HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4.27 min. Example 201 (General procedure (C)) 5-(3-Bromo-4-methoxybenzylidene)thiazolidine-2,4-dione O CH 3 HN 'aBr 20 O HPLC-MS (Method A): m/z: 314 (M+1), Rt = 3.96 min. Example 202 (General procedure (C)) 3-{(2-Cyanoethyl)-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]amino)propionitrile WO 2004/080480 PCT/DK2004/000158 159 N HN I o N HPLC-MS (Method A): m/z: 327 (M+1); Rt = 2.90 min. Example 203 (General procedure (C)) 5 3 -(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid methyl ester o H N HN ~9\0 O
O-CH
3 HPLC-MS (Method A): m/z: 303 (M+1); Rt = 3.22-3-90 min. Example 204 10 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid pentyl ester. O H HN 0 0 0 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid methyl ester (example 203, 59 mg; 0.195mmol) was stirred in pentanol (20 mL) at 145 °C for 16 hours. The mixture was evaporated to dryness affording the title compound (69 mg). 15 HPLC-MS (Method C): m/z: 359 (M+1); Rt.= 4.25 min. Example 205 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-7-carboxylic acid O HHO HNO 20 0 HPLC-MS (Method A): m/z: 289 (M+1); Rt = 2.67 min. Example 206 (General procedure (C)) 5-(1 -Benzylindol-3-ylmethylene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 160 0 S N HN 0 HPLC-MS (Method A): m/z: 335 (M+1); Rt = 4.55 min. Example 207 (General procedure (C)) 5 5-(1-Benzenesulfonylindol-3-ylmethylene)thiazolidine.-2,4-dione N H NO o N° HPLC-MS (Method A): m/z: = 385 (M+1); Rt = 4.59 min. Example 208 (General procedure (C)) 10 5-(4-[1,2,3]Thiadiazol-4-ylbenzylidene)thiazolidine-2,4-dione 0 N:N HN . I . O 0 HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.45 min. Example 209 (General procedure (C)) 15 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dione Q+ OO 0 HN K 0 HPLC-MS (Method A): m/z: 357 (M+1); Rt - 4.42 min.
WO 2004/080480 PCT/DK2004/000158 161 Example 210 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-1l-carboxylic acid ethyl ester oH 3 CvO-€O '-s NN HN I I 0 HPLC-MS (Method A): m/z: 317 (M+1); Rt = 4.35 min. 5 Example 211 (General procedure (C)) 5-[2-(4-Pentylbenzoyl)-benzofuran-5-ylmethylene]thiazolidine-2,4-dione o HN _ I
CH
3 O HPLC-MS (Method A): m/z: 420 (M+1); Rt = 5.92 min. 10 Example 212 (General procedure (C)) 5-[1-(2-Fluorobenzyl)-4-nitroindol-3-ylmethylene]thiazolidine-2,4-dione F HN O% HPLC-MS (Method A): (Anyone 1) m/z: 398 (M+1); Rt = 4.42 min. 15 Example 213 (General procedure (C)) 5-(4-Benzyloxyindol-3-ylmethylene)thiazolidine-2,4-dione O H S N O HN ~I 0 -0 6 HPLC-MS (Method A): m/z: 351 (M+1); Rt = 3.95 min. 20 WO 2004/080480 PCT/DK2004/000158 162 Example 214 (General procedure (C)) 5-(4-Isobutylbenzylidene)-thiazolidine-2,4-dione
H
3 C CH 3 0 HPLC-MS (Method A): m/z: 262 (M+1); Rt = 4.97 min. 5 Example 215 (General procedure (C)) Trifluoromethanesulfonic acid 4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yl ester F F o OO HN O HPLC-MS (Method A): m/z: 404 (M+1); Rt = 4.96 min. 10 Preparation of starting material: 4-Hydroxy-1-naphthaldehyde (10 g, 58 mmol) was dissolved in pyridin (50 ml) and the mix ture was cooled to 0-5 °C. With stirring, trifluoromethanesulfonic acid anhydride (11.7 ml, 70 mmol) was added drop-wise. After addition was complete, the mixture was allowed to warm 15 up to room temperature, and diethyl ether (200 ml) was added. The mixture was washed with water (2 x 250 ml), hydrochloric acid (3N, 200 ml), and saturated aqueous sodium chloride (100 ml). After drying (MgSO4), filtration and concentration in vacuo, the residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 8.35 g (47%) trifluoromethanesulfonic acid 4-formylnaphthalen-1-yl ester, 20 mp 44-46.6 'C. Example 216 (General procedure (C)) 5-(4-Nitroindol-3-ylmethylene)-thiazolidine-2,4-dione O H HN 25 HPLC-MS (Method A): moz: 290 (M+1); Rt = 3.14 m. 25 HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.14 min.
WO 2004/080480 PCT/DK2004/000158 163 Example 217 (General procedure (C)) 5-(3,5-Dibromo-4-hydroxy-benzylidene)thiazolidine-2,4-dione H Br OOH HN I B 0 5 1 H NMR (DMSO-d 6 ): 6 H = 12.65 (broad,1H), 10.85 (broad,1H), 7.78 (s,2H), 7.70 (s,1H); HPLC-MS (Method A): m/z: 380 (M+1); Rt = 3.56 min. Example 218 (General procedure (C)) O HNN 0 10 HPLC-MS (Method A): m/z: 385 (M+1); Rt = 5.08 min. General procedure for preparation of starting materials for examples 218 - 221: Indole-3-carbaldehyde (3.8 g, 26 mmol) was stirred with potassium hydroxide (1.7 g) in ace tone (200 mL) at RT until a solution was obtained indicating full conversion to the indole po 15 tassium salt. Subsequently the solution was evaporated to dryness in vacuo. The residue was dissolved in acetone to give a solution containing 2.6 mmol/20 mL. 20 mL portions of this solution were mixed with equimolar amounts of arylmethylbromides in acetone (10 mL). The mixtures were stirred at RT for 4 days and subsequently evaporated to 20 dryness and checked by HPLC-MS. The crude products, 1-benzylated indole-3 carbaldehydes, were used for the reaction with thiazolidine-2,4-dione using the general pro cedure C. Example 219 (General procedure (C)) 25 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-ylmethyl]benzoic acid methyl ester WO 2004/080480 PCT/DK2004/000158 164 -- cH, HN HPLC-MS (Method A): m/z: 393 (M+1); Rt = 4.60 min. Example 220 (General procedure (C)) 5 5-[1 -(9,10-Dioxo-9,10-dihydroanthracen-2-ylmethyl)-1 H-indol-3-ylmethylene]thiazolidine-2,4 dione o HN 0 0 HPLC-MS (Method A): m/z: 465 (M+1); Rt = 5.02 min. 10 Example 221 (General procedure (C)) 4'-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-ylmethyl]biphenyl-2-carbonitrile N HNN 0 HPLC-MS (Method A): m/z: 458 (M+23); Rt = 4.81 min. 15 Example 222 (General procedure (C)) 3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-methylindol-1 -ylmethyl]benzonitrile. H S H C p N O N / 2-Methylindole-3-carbaldehyde (200 mg, 1.26 mmol) was added to a slurry of 3 bromomethylbenzenecarbonitrile (1.26 mmol) followed by sodium hydride, 60%, (1.26 mmol) 20 in DMF (2 mL). The mixture was shaken for 16 hours, evaporated to dryness and washed WO 2004/080480 PCT/DK2004/000158 165 with water and ethanol. The residue was treated with thiazolidine-2,4-dione following the general procedure C to afford the title compound (100 mg). HPLC-MS (Method C): m/z: 374 (M+1); Rt. = 3.95 min. 5 Example 223. (General procedure (C)) 5-(1-Benzyl-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione. O 3C HNO 0 This compound was prepared in analogy with the compound described in example 222 from 10 benzyl bromide and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine 2,4-dione resulting in 50 mg of the title compound. HPLC-MS (Method C): m/z: 349 (M+1); Rt. = 4.19 min. 15 Example 224 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-methylindol-1 -ylmethyl]benzoic acid methyl es ter CH, O H3C S N ~I HN 0 1 This compound was prepared in analogy with the compound described in example 222 from 20 4-(bromomethyl)benzoic acid methyl ester and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine-2,4-dione. HPLC-MS (Method C): m/z: 407 (M+1); Rt.= 4.19 min.
WO 2004/080480 PCT/DK2004/000158 166 Example 225 (General procedure (C)) 5-(2-Chloro-1-methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione O CH HN C N
HN--
0 HPLC-MS (Method A): m/z: 293 (M+1); Rt = 4.10 min. 5 Example 226 (General procedure (C)) 5-(4-Hydroxy-3,5-diiodo-benzylidene)-thiazolidine-2,4-dione O H S OH 01 HN O HPLC-MS (Method A): m/z: 474 (M+1); Rt = 6.61 min. 10 Example 227 (General procedure (C)) 5-(4-Hydroxy-3-iodobenzylidene)thiazolidine-2,4-dione 0 .~OH HN " 1 0 HPLC-MS (Method C): m/z: 348 (M+1); Rt. = 3.13 min 15 1 H-NMR: (DMSO-d 6 ): 11.5 (1H,broad); 7.95(1H,d); 7.65(1H,s); 7.45 (1H,dd); 7.01(1H,dd); 3.4 (1H,broad). Example 228 (General procedure (C)) 20 5-(2,3,6-Trichlorobenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 167 OCI S C , HN 0 Cl H PLC-MS (Method C): m/z: 309 (M+1); Rt.= 4.07 min 5 Example 229 (General procedure (C)) 5-(2,6-Dichlorobenzylidene)thiazolidine-2,4-dione 0 -S HN O CI Mp. 152-154 0 C. HPLC-MS (Method C): m/z: 274 (M+1), Rt.= 3.70 min 10 1 H-NMR: (DMSO-do): 12.8 (1H, broad); 7.72 (1H,s); 7.60 (2H,d); 7.50 (1H,t). Example 230 (General procedure (C)) 5-[1-(2,6-Dichloro-4-trifluoromethylphenyl)-2,5-dimethyl-1 H-pyrrol-3-ylmethylene]thiazolidine 2,4-dione FF F C H-N - /CH 3 15 0 HPLC-MS (Method C): m/z: 436 (M+1); Rt. 4.81 min Example 231 (General procedure (C)) 5-[1-(3,5-Dichlorophenyl)-5-(4-methanesulfonylphenyl)-2-methyl-1H-pyrrol-3-ylmethylene] 20 thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 168 cil 0 H3 c HN N O o cH 3 HPLC-MS (Method C): m/z: 508 (M+1); Rt. = 4.31 min Example 232 (General procedure (C)) 5 5-[1 -(2,5-Dimethoxyphenyl)-5-(4-methanesulfonylphenyl)-2-methyl-1H-pyrrol-3-ylmethylene] thiazolidine-2,4-dione
H
3 C, OCH 3 o H3C - o HN O/ O
CH
3 HPLC-MS (Method C): m/z: 499 (M+1); Rt. = 3.70 min 10 Example 233 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2,5-dimethylpyrrol-1-yl]benzoic acid HO 0 0 ; SH3C Ns HN CH3 0 HPLC-MS (Method C): m/z:342 (M+1); Rt.= 3.19 min 15 Example 234 (General procedure (C)) 5-(4-Hydroxy-2,6-dimethoxybenzylidene)thiazolidine-2,4-dione CH3 O H H OOH 0,'CH3 HPLC-MS (Method C): m/z:282( M+1); Rt.= 2.56, mp=331-333 "C 20 WO 2004/080480 PCT/DK2004/000158 169 Example 235 (General procedure (C)) 5-(2,6-Dimethylbenzylidene)thiazolidine-2,4-dione HN O
CH
3 M.p: 104-105 °C 5 HPLC-MS (Method C): m/z: 234 (M+1); Rt.= 3.58 min, Example 236 (General procedure (C)) 5-(2,6-Dimethoxybenzylidene)thiazolidine-2,4-dione HN o
OCH
3 10 Mp: 241-242 OC HPLC-MS (Method C): m/z: 266 (M+1); Rt.= 3.25 min; Example 237 (General procedure (C)) 5-[4-(2-Fluoro-6-nitrobenzyloxy)-2,6-dimethoxybenzylidene]thiazolidine-2,4-dione 0 H 3 F HN C 15
CH
3 Mp: 255-256 C HPLC-MS (Method C): m/z: 435 (M+1), Rt 4.13 min, Example 238 (General procedure (C)) 20 5-Benzofuran-2-ylmethylenethiazolidine-2,4-dione 00 HON 0 HPLC-MS (Method C): m/z:246 (M+1); Rt.= 3.65 min, mp =265-266 *C.
WO 2004/080480 PCT/DK2004/000158 170 Example 239 (General procedure (C)) 5-[3-(4-Dimethylaminophenyl)allylidene]thiazolidine-2,4-dione 0 YH 3 ON H HN N, ' C H 0 5 HPLC-MS (Method C): m/z:276(M+1); Rt.= 3.63, mp = 259-263 'C 'H-NMR: (DMSO-d) 6= 12.3 (1H,broad); 7.46 (2H,d); 7.39 (1H,d); 7.11 (1H,d); 6.69 (2H,d); 6.59 (1H, dd); 2.98 (3H,s). Example 240 (General procedure (C)) 10 5-(2-Methyl-3-phenylallylidene)thiazolidine-2,4-dione 0 HN O H 3 HN 0 Mp: 203-210 'C HPLC-MS (Method C): m/z: 246 (M+1); Rt = 3.79 min. 15 Example 241 (General procedure (C)) 5-(2-Chloro-3-phenylallylidene)thiazolidine-2,4-dione HN N 0 O Mp: 251-254 °C HPLC-MS (Method C): m/z: 266 (M+1; Rt = 3.90 min 20 Example 242 (General procedure (C)) 5-(2-Oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione H 02 0 N HN -. 0 25 Mp: 338-347 *C WO 2004/080480 PCT/DK2004/000158 171 HPLC-MS (Method C): m/z: 273 (M+1); Rt. = 2.59 min. Example 243 (General procedure (C)) 5-(2,4,6-Tribromo-3-hydroxybenzylidene)thiazolidine-2,4-dione. O OH HN Br zBr HN Br 5 O Br HPLC-MS (Method C): m/z: 459 (M+1);Rt.= 3.65 min. Example 244 (General procedure (C)) 5-(5-Bromo-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione. 0\ H C H HN 0 10 Br HPLC-MS (Method C): mlz: 339 (M+1); Rt = 3.37min. Example 245 (General procedure (C)) 5-(7-Bromo-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione. 0H 15 Br HN 15 HPLC-MS (Method C): m/z: 319 (M+1); Rt = 3.48min. Example 246 (General procedure (C)) 5-(6-Bromoindol-3-ylmethylene)thiazolidine-2,4-dione. HN Br 20 o HPLC-MS (Method C): m/z: 325 (M+1); Rt = 3.54 min. Example 247 (General procedure (C)) 5-(8-Methyl-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
WO 2004/080480 PCT/DK2004/000158 172 S H CH 3 O HN O N. HPLC-MS (Method C): m/z: 287 (M+1); Rt = 2.86 min. 5 Example 248 (General procedure (C)) 5-(6-Methoxy-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione. O H 0. O_ N. A O
CH
3 HPLC-MS (Method C): m/z: 303 (M+1); Rt = 2.65 min. 10 Example 249 (General procedure (C)) 5-Quinolin-3-ylmethylenethiazolidine-2,4-dione. 0 : s ,N HN o HPLC-MS (Method C): m/z: 257 (M+1); Rt = 2.77 min. 15 Example 250 (General procedure (C)) 5-(8-Hydroxyq uinolin-2-ylmethylene)thiazolidine-2,4-dione. 0 S HN O OH HPLC-MS (Method C): m/z: 273 (M+1); Rt = 3.44 min. 20 Example 251 (General procedure (C)) 5-Quinolin-8-ylmethylenethiazolidine-2,4-dione. HN (M. 0 N, HPLC-MS (Method C): m/z: 257 (M+1); Rt = 3.15 min.
WO 2004/080480 PCT/DK2004/000158 173 Example 252 (General procedure (C)) 5-(1-Bromo-6-methoxynaphthalen-2-ylmethylene)thiazolidine-2,4-dione. 0 HN O CH3 O Br 5 HPLC-MS (Method C): m/z: 366 (M+1); Rt = 4.44 min. Example 253 (General procedure (C)) 5-(6-Methyl-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione. O H HN CH 0 10 HPLC-MS (Method C): m/z: 287 (M+1); Rt. = 2.89 min. Example 254 (General procedure (D)) 5-(2,6-Dichloro-4-dibenzylaminobenzylidene)thiazolidine-2,4-dione. N -N HN I o CI 15 HPLC-MS (Method C): m/z: 469 (M+1); Rt = 5.35 min. Example 255 (General Procedure (C)) 7
-(
2 ,4-Dioxothiazolidin-5-ylidenemethyl)-4-methoxybenzofuran-2-carboxylic acid O CH 3 S Op 0- 0 HN o 0 HO 20 HPLC-MS (Method C): m/z: 320 (M+1); Rt = 2.71 mln. Preparation of the intermediate, 7-formyl-4-methoxybenzofuran-2-carboxylic acid: WO 2004/080480 PCT/DK2004/000158 174 A mixture of 2-hydroxy-6-methoxybenzaldehyde (6.4 g, 42 mmol), ethyl bromoacetate (14.2 mL, 128 mmol) and potassium carbonate (26 g, 185 mmol) was heated to 130 oC. After 3 h the mixture was cooled to room temperature and acetone (100 mL) was added, the mixture was subsequently filtered and concentrated in vacuo. The residue was purified by column 5 chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 7.5 g (55%) of ethyl 4-methoxybenzofuran-2-carboxylate. A solution of ethyl 4-methoxybenzofuran-2-carboxylate (6.9 g, 31.3 mmol) in dichloro methane (70 ml) was cooled to 0 OC and a solution of titanium tetrachloride (13.08 g, 69 10 mmol) was added drop wise. After 10 minutes dichloromethoxymethane (3.958 g, 34 mmol) was added over 10 minutes. After addition, the mixture was warmed to room temperature for 18 hours and the mixture poured into hydrochloric acid (2N, 100 mL). The mixture was stirred for 0.5 hour and then extracted with a mixture of ethyl acetate and toluene (1:1). The organic phase was dried over Na 2
SO
4 and concentrated in vacuo. The residue was purified by col 15 umn chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 5.8 g (80%) of ethyl 7-formyl-4-methoxybenzofuran-2-carboxylate. 7-formyl-4-methoxybenzofuran-2-carboxylate (5.0 g, 21.5 mmol) and sodium carbonate (43 mmol) in water (100 mL) was refluxed until a clear solution appeared (about 0.5 hour). The 20 solution was filtered and acidified to pH =1 with hydrochloric acid (2 N), the resulting product was filtered off and washed with ethyl acetate and ethanol and dried to afford 3.5 g (74%) of 7-formyl-4-methoxybenzofuran-2-carboxylic acid as a solid. 1 H NMR (DMSO-d 6 ): 6= 10.20 (s, 1H); 8.07 (d, 1H); 7.70 (s, 1H); 7.17 (d, 1H); 4.08 (s, 25 3H). Example 256 (General Procedure (C)) 5-(4-Methoxybenzofuran-7-ylmethylene)thiazolidine-2,4-dione HN 3 CH 30 HPLC-MS (Method C): mz: 267 (M+1); Rt = 3.30 m. 30 HPLC-MS (Method C): m/z: 267 (M+1); Rt = 3.30 main.
WO 2004/080480 PCT/DK2004/000158 175 Preparation of the intermediate, 4-methoxybenzofuran-7-carbaldehyde: A mixture of 7-formyl-4-methoxybenzofuran-2-carboxylic acid (3.0 g, 13.6 mmol) and Cu (0.6 g, 9.44 mmol) in quinoline (6 mL) was refluxed. After 0.5 h the mixture was cooled to room 5 temperature and water (100 mL) and hydrochloric acid (10 N, 20 mL) were added. The mix ture was extracted with a mixture of ethyl acetate and toluene (1:1), filtered through celite and the organic layer separated and washed with a sodium carbonate solution, dried over Na 2
SO
4 and concentrated in vacuo to afford 1.5 g crude product. Column chromatography SiO 2 , EtOAc/heptanes=1/4 gave 1.1 g (46%) of 4-methoxybenzofuran-7-carbaldehyde as a 10 solid. 1 H NMR (CDC3): 6:10.30 (s,1H) ; 7.85 (d,1H) ; 7.75 (d,1H); 6.98 (d,1H); 6.87 (d,IH); 4.10 (s,3H). HPLC-MS (Method C) :m/z: 177 (M+1); Rt. = 7.65 min. 15 Example 257 (General Procedure (C)) 5-(4-Hydroxybenzofuran-7-ylmethylene)thiazolidine-2,4-dione 0 OS /OH HN O O HPLC-MS (Method C): m/z: = 262 (M+1); Rt 2.45 min. 20 Preparation of the intermediate, 4-hydroxybenzofuran-7-carbaldehyde A mixture of 4-methoxybenzofuran-7-carbaldehyde (1.6 g, 9.1 mmol) and pyridine hydrochlo ride (4.8 g, 41.7mmol) in quinoline (8 mL) was refluxed. After 8 h the mixture was cooled to room temperature and poured into water (100 mL) and hydrochloric acid (2 N) was added to 25 pH = 2. The mixture was extracted with a mixture of ethyl acetate and toluene (1:1), washed with a sodium carbonate solution, dried with Na 2
SO
4 and concentrated in vacuo to afford 0.8 g crude product. This was purified by column chromatography on silica gel, eluting with a mixture of ethyl acetate and heptane (1:3). This afforded 250 mg of 4-hydroxybenzofuran-7 carbaldehyde as a solid. 30 WO 2004/080480 PCT/DK2004/000158 176 1 H NMR (DMSO-d 6 ): 6= 11.35 (s, broad,1H); 10.15 (s, 1H); 8.05 (d, 1H) ; 7.75 (d, 1H); 7.10 (d, 1H); 6.83 (d, 1H). HPLC-MS (Method C): m/z: 163 (M+1); Rt. = 6.36 min. Example 258 (General Procedure (C)) 5 5-(5-Bromo-2,3-dihydrobenzofuran-7-ylmethylene)thiazolidine-2,4-dione Br S/ HN O O HPLC-MS (Method C): m/z: 328 (M+1); Rt = 3.66 min. Preparation of the intermediate, 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde: 10 To a cooled (15 OC) stirred mixture dihydrobenzofuran (50.9 g, 0.424 mol) in acetic acid (500 mL), a solution of bromine (65.5 mL, 1.27 mol) in acetic acid (200 mL) was added drop wise over 1 hour. After stirring for 18 hours, a mixture of Na 2
S
2 Os (150 g) in water (250 mL) was added carefully, and the mixture was concentrated in vacuo. Water (200 mL) was added and the mixture was extracted with ethyl acetate containing 10% heptane, dried over Na 2
SO
4 and 15 concentrated in vacuo to give crude 5,7-dibromo-2,3-dihydrobenzofuran which was used as such for the following reaction steps. To a cooled solution (-78 OC) of crude 5,7-dibromo-2,3 dihydrobenzofuran (50.7 g, 0.182 mol) in THF (375 mL) a solution of n-BuLi (2.5 M, 80 mL, 0.200 mol) in hexane was added. After addition, the mixture was stirred for 20 min. DMF (16 mL) was then added drop wise at -78 OC. After addition, the mixture was stirred at room tem 20 perature for 3 h and then the mixture was poured into a mixture of ice water, (500 mL) and hydrochloric acid (10 N, 40 mL) and extracted with toluene, dried over Na 2
SO
4 and concen trated in vacuo. Column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4) afforede 23 g of 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde as a solid. 25 1 H NMR (CDCl 3 ): 6:10.18 (s, 1H) ; 7.75 (d,1H) ;7.55 (d,1H) ; 4.80 (t,2H) ; 3.28 (t,2H). Example 259 (General Procedure (C)) 5-(4-Cyclohexylbenzylidene)thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 177 O S HN O HPLC-MS (Method C): m/z: 288 (M+1); Rt = 5.03 min. Preparation of the intermediate, 4-cyclohexylbenzaldehyde: 5 This compound was synthesized according to a modified literature procedure (J. Org. Chem., 37, No.24, (1972), 3972-3973). Cyclohexylbenzene (112.5 g, 0.702 mol) and hexamethylenetetramine (99.3 g, 0.708 mol) were mixed in TFA (375 mL). The mixture was stirred under nitrogen at 90 oC for 3 days. Af 10 ter cooling to room temperature the red-brown mixture was poured into ice-water (3600 ml) and stirred for 1 hour. The solution was neutralized with Na 2
CO
3 (2 M solution in water) and extracted with dichloromethane (2.5 L). The organic phase was dried (Na 2
SO
4 ) and the sol vent was removed in vacuo. The remaining red-brown oil was purified by fractional distillation to afford the title compound (51 g, 39%). 15 1 H NMR (CDCl 3 ): 89.96 (s, 1H), 7.80 (d, 2H), 7.35 (d, 2H), 2.58 (m, 1H), 1.94-1.70 (m, 5 H), 1.51-1.17 (m, 5H) Other ligands of the invention include 20 3',5'-Dichloro-4'-(2,4-dioxothiazolidin-5-ylidenemethyl)biphenyl-4-carboxylic acid: 0 O O>SC | OH S CI\ HN O Cl Example 260 (General procedure (C)) 5-(1-Bromo-6-hydroxynaphthalen-2-ylmethylene)-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 178 O HN
-
OH O Br HPLC-MS (Method C): m/z = 350 (M+1); Rt. = 3.45 min. Example 261 (General procedure (C)) 5 5-[4-(2-Bromoethoxy)-naphthalen-1 -ylmethylene]-thiazolidine-2,4-dione Br 0 S O HN o HPLC-MS (Method C): m/z = 380 (M+1); Rt = 3.52 min. Example 262 (General procedure (C)) 10 5-(2-Methyl-5-nitro-1H-indol-3-ylmethylene)-thiazolidine-2,4-dione 0 H N 0 CH3 N H HPLC-MS (Method C): m/z = 304 (M+1); Rt = 2.95 min. Example 263 (General procedure (C)) 15 5-(4-Naphthalen-2-yl-thiazol-2-ylmethylene)-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 179 H I ONO HI/ s S , IN HPLC-MS (Method C): m/z = 339 (M+1); Rt.= 4.498 min. Example 264 (General procedure (C)) 5 5-[4-(4-Methoxy-naphthalen-1 -yl)-thiazol-2-ylmethylene]-thiazolidine-2,4-dione H o N-fO Is NO
H
3
C
HPLC-MS (Method C): m/z = 369 (M+1); Rt.= 4.456 min. Example 265 (General procedure (C)) 10 5-(2-Pyridin-4-yl-1H-indol-3-ylmethylene)-thiazolidine-2,4-dione O H H N NN H HPLC-MS (Method C): m/z = 322 (M+1); Rt. = 2.307 min.
WO 2004/080480 PCT/DK2004/000158 180 Example 266 (General procedure (C)) 5-[5-(4-Chlorophenyl)-1 H-pyrazol-4-ylmethylene]-thiazolidine-2,4-dione O HN H O s N H CI HPLC-MS (Method C): m/z = 306 (M+1); Rt.= 3.60 min. 5 Example 267 (General procedure (C)) 5-[5-(2,5-Dimethylphenyl)-1 H-pyrazol-4-ylmethylene]-thiazolidine-2,4-dione 0O H
CH
3
N
H H S O0
CH
3 N-N H HPLC-MS (Method C): m/z = 300 (M+1); Rt. = 3.063 min. 10 Example 268 (General procedure (C)) 5-(2-Phenyl-benzo[d]imidazo[2,1-b]thiazol-3-ylmethylene)-thiazolidine-2,4-dione s N N 0 N I H HPLC-MS (Method C): m/z = 378 (M+1); Rt = 3.90 min. 15 WO 2004/080480 PCT/DK2004/000158 181 Example 269 (General procedure (C)) N-{4-[2-(2,4-Dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-acetamide 0HC O -NH H 3 C 0 0 ~O 0 HPLC-MS (Method C): m/z = 355 (M+1); Rt 3.33 min. 5 Example 270 (General procedure (C)) 5-(2-Phenyl-imidazo[1,2-a]pyridin-3-ylmethylene)-thiazolidine-2,4-dione O 'H N S N HPLC-MS (Method C): m/z = 322 (M+1); Rt. = 2.78 min. 10 Example 271 (General procedure (C)) 5-(2-Naphthalen-2-yl-imidazo[1,2-a]pyridin-3-ylmethylene)-thiazolidine-2,4-dione H 0 N / N HPLC-MS (Method C): m/z = 372 (M+1); Rt. = 2.78 min. 15 Example 272 (General procedure (C)) WO 2004/080480 PCT/DK2004/000158 182 5-[6-Bromo-2-(3-methoxyphenyl)-imidazo[1,2-a]pyridin-3-ylmethylene]-thiazolidine-2,4-dione
O-OH
3 N Br SN \/ S O0 N O H HPLC-MS (Method C): m/z = 431 (M+1); Rt.= 3.30 min. 5 Example 273 (General procedure (C)) 5-(1,2,3,4-Tetrahydrophenanthren-9-ylmethylene)thiazolidine-2,4-dione H 10 S N, H 0 HPLC-MS (Method C): m/z = 310 (M+1); Rt.= 4.97 min. 10 Example 274 (General procedure (C)) 5-(3,5, 5,8,8-Pentamethyl-5,6,7,8-tetrahydro-naphthalen-2-ylmethylene)thiazolidine-2,4-dione H O" 3 3 H CH3 H3C CH 3 HPLC-MS (Method C): m/z = 330 (M+1); Rt.= 5.33 min.
WO 2004/080480 PCT/DK2004/000158 183 Example 275 (General procedure (C)) 5-[6-(2,4-Dichloro-phenyl)-imidazo[2, 1-b]thiazol-5-ylmethylene]-thiazolidine-2,4-dione CI N -S ClN H 0O;4,N 0 H HPLC-MS (Method C): mlz = 396 (M+1); Rt. = 3.82 min. 5 Example 276 (General procedure (C)) 5-(5-Bromobenzofuran-7-ylmethylene)-thiazolidine-2,4-dione HN Br O HPLC-MS (Method C): m/z = 324 (M+1); Rt. = 3.82 min. 10 Example 277 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-1,4-dimethylcarbazol-9-ylmethyl]-benzoic acid H 0 N N OS
CH
3
H
3 C NoO OH HPLC-MS (Method C): m/z = 457 (M+1); Rt = 4,23 min. 15 WO 2004/080480 PCT/DK2004/000158 184 Preparation of intermediary aldehyde: 1,4 Dimethylcarbazol-3-carbaldehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mix ture was stirred for 1 hour at room temperature. 4-Bromomethylbenzoic acid (0.73 g, 3.4 5 mmol) was slowly added and the resulting slurry was heated to 40 °C for 16 hours. Water (5 mL) and hydrochloric acid (6N, 3 mL) were added. After stirring for 20 min at room tempera ture, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formyl-1,4-dimethylcarbazol-9-ylmethyl)benzoic acid. 10 HPLC-MS (Method C): m/z = 358 (M+1), RT. = 4.15 min. Example 278 (General procedure (C)) 4-[7-(2,4-Dioxothiazolidin-5-ylidenemethyl)-benzofuran-5-yl]-benzoic acid S HN O\ O 15 Starting aldehyde commercially available (Syncom BV, NL) HPLC-MS (Method C): m/z = 366 (M+1); Rt. = 3.37 min. Example 279 (General procedure (C)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2-nitrophenoxy]-benzoic acid methyl ester 3H C 0 0 H3c"O O HN
N:
O IO O 20 HPLC-MS (Method C): m/z 401 (M0); Rt. = 4.08 in. 20 HPLC-MS (Method C): mlz = 401 (M+1); Rt. = 4.08 min.
WO 2004/080480 PCT/DK2004/000158 185 Example 280 (General procedure (C)) 3',5'-Dichloro-4'-(2,4-dioxothiazolidin-5-ylidenemethyl)-biphenyl-4-carboxylic acid OH HN O Cl 5 Starting aldehyde commercially available (Syncom BV, NL) HPLC-MS (Method C): m/z = 394 (M+1); Rt. = 3.71 min. Example 281 (General procedure (C)) 0 HN s 10 o HPLC-MS (Method C): m/z = 232( M+1); Rt.= 3.6 min. Example 282. 5-(2-Methyl-1 H-indol-3-ylmethyl)-thiazolid ine-2,4-dione 0 H N SxO CH3 N 15 H 5-(2-Methyl-lH-indol-3-ylmethylene)thiazolidine-2,4-dione (prepared as described in example 187, 1.5 g, 5.8 mmol) was dissolved in pyridine (20 mL) and THF (50 mL), LiBH 4 (2 M in THF, 23.2 mmol) was slowly added with a syringe under cooling on ice. The mixture was heated to 85 °C for 2 days. After cooling, the mixture was acidified with concentrated hydro- WO 2004/080480 PCT/DK2004/000158 186 chloric acid to pH 1. The aquous layer was extracted 3 times with ethyl acetate, dried with MgSO 4 treated with activated carbon, filtered and the resulting filtrate was evaporated in vacuo to give 1.3 g (88%) of the title compound. 5 HPLC-MS (Method C): m/z = 261 (M+1); Rt. = 3.00 min. Example 283 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyric acid 0 HN 0 10 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyric acid (4.98 g, 13.9 mmol, prepared as described in example 469) was dissolved in dry THF (50 mL) and added dry pyridine (50 mL) and, in portions, lithium borohydride (2.0 M, in THF, 14 mL). The result ing slurry was refluxed under nitrogen for 16 hours, added (after cooling) more lithium boro hydride (2.0 M, in THF, 7 mL). The resulting mixture was refluxed under nitrogen for 16 15 hours. The mixture was cooled and added more lithium borohydride (2.0 M, in THF, 5 mL). The resulting mixture was refluxed under nitrogen for 16 hours. After cooling to 5 OC, the mix ture was added water (300 mL) and hydrochloric acid (150 mL). The solid was isolated by filtration, washed with water (3 x 500 mL) and dried. Recrystallization from acetonitrile (500 mL) afforded2.5 g of the title compound. 20 1 H-NMR (DMSO-d 6 , selected peaks): 8 = 3.42 (1H, dd), 3.90 (1H, dd), 4.16 (2H, "t"), 4.95 (1H, dd), 6.92 (1H, d), 7.31 (1H, d), 7.54 (1H, t), 7.62 (1H, t), 8.02 (1H, d), 8.23 (1H, d), 12.1 (1 H, bs), 12.2 (1 H, bs). HPLC-MS (Method C): m/z = 382 (M+23); Rt = 3,23 min. 25 Example 284 5-Naphthalen-1 -ylmethylthiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 187 o S HN 0 5-Naphthalen-1-ylmethylenethiazolidine-2,4-dione (1.08 g, 4.2 mmol, prepared as described in example 68) was dissolved in dry THF (15 mL) and added dry pyridine (15 mL) and, in portions, lithium borohydride (2.0 M, in THF, 4.6 mL). The resulting mixture was refluxed un 5 der nitrogen for 16 hours. After cooling to 5 OC, the mixture was added water (100 mL), and, in portions, concentrated hydrochloric acid (40 mL). More water (100 mL) was added, and the mixture was extracted with ethyl acetate (200 mL). The organic phase was washed with water (3 x 100 mL), dried and concentrated in vacuo. The residue was dissolved in ethyl ace tate (50 mL) added activated carbon, filtered and concentrated in vacuo and dried to afford 10 0.82 g (75%) of the title compound. 1 H-NMR (DMSO-d 6 ): 8 = 3.54 (1H, dd), 3.98 (1H, dd), 5.00 (1H, dd), 7.4-7.6 (4H, m), 7.87 (1H, d), 7.96 (1H, d), 8.11 (1H, d), 12.2 (1H, bs). HPLC-MS (Method C): m/z = 258 (M+1); Rt = 3,638 min. 15 The following preferred compounds of the invention may be prepared according to proce dures similar to those described in the three examples above: Example 285 H o 0 Example 286 HO O H N Example 287 O HNS CH 3 OBr 0 WO 2004/080480 PCT/DK2004/000158 188 Example 288 0 HN S CH 3 0 Example 289 0 Example 290 0. HN 0 Example 291 O 0 SO ~~JOH HN Br 0 Example 292 0 HNH 0 Example 293 0 HN NH 0 Example 294 0 HN 0 CH 3 Example 295 0 H HN j O CH3 Example 296 0 HN 0 Br WO 2004/080480 PCT/DK2004/000158 189 Example 297 O0 H -s N HN o Example 298 CH H N 0 O CH Example 299 HC H '-s HN O
CH
3 Example 300 O I HN O Example 301 O HN H Example 302 o
OH
3 HN S 4CH 3 0 Example 303 0 HNS CH 3 0 Example 304 HN S OH
O
WO 2004/080480 PCT/DK2004/000158 190 Example 305 Br o CH 3 o HN I 0 Example 306 cl I cl o Example 307 o o N HN 0 Example 308 HN O 0 0 Example 309 o Example 310 0 S N HN o Example 311 o OH HNO CH 0 Example 312 0 HN I N 0 CH 3 WO 2004/080480 PCT/DK2004/000158 191 Example 313 o H HN 0 o Example 314 O H 3 C -s HN 0 Example 315 N HN N 0N Example 316 O H N HN 0 O
O-CH
3 Example 317 O H HO H S N O HiN 0 Example 318 O - N HN 0 Example 319 o I. o-o 00 HN O 0 Example 320 OH 3 CO O 0 WO 2004/080480 PCT/DK2004/000158 192 Example 321 o 0 HN 0 1CH 0 Example 322 O OH s S O 10 N HN I 0 Example 323
CH
3 0 HN 0 Example 324 HN I 0 O Example 325 O OH H NS~ O O--NA HNS 1 0 O 0 Example 327 H O OF HN 0 Example 328 o HN0O WO 2004/080480 PCT/DK2004/000158 193 0 Example 329 o o oO 0 Example 330 0 o HNS 0 0 Example 331 o O HN H O 0 Example 332 HN OH Br o Example 333 0 S N Br HN 0 Example 334 0 HN o ci Example 335 0 H HN N Br OOH Example 336 O CH3
HNI
O H 0
CH
3 WO 2004/080480 PCT/DK2004/000158 194 Example 337 O HN O
CH
3 Example 338 CH O 0 H SN HN N -- O Example 339 H HN 0 Example 340 0 CH 0 H H Example 342 O0 -s o -N. OH ExampHeN34O H HN 0,. NI "N"o O OCH3 Example 343 0 HN oHNH O Example 344 CH3 HN OH 0 0 HN
CH
3 0 OH 3 WO 2004/080480 PCT/DK2004/000158 195 Example 345 O Cl HN O Cl Example 346 o GH S 0 HN O O
OCH
3 Example 347 0 ~'s HN O O-CH 3 Example 348 O F HN o F Example 349 0 CH 3 HN 0 CH 3 Example 350 o HN O 0' CH o Br Example 351 0 H HN CH CH, 0 Example 352
CH
3 0 \ H 3 C N HN 0 WO 2004/080480 PCT/DK2004/000158 196 Example 353 O0 HN S Cl Br O cl Example 354 0 Example 355 0~ ~ "H HN O Example 355 0 S O' CH3 HN 0 O OOH Example 357 Br . o HN o 0 Example 357 Br HN 0 n Exam ple 358 mo o o> E x a m p le 3 5 9 o o H A s 0 0 0 WO 2004/080480 PCT/DK2004/000158 197 Example 360 0 HN _s rcH 3 / OH O N 0 IHOH Example 361 HN 0 I OH 0 Example 362 O HN Br 0 Example 363 0 HN
-
OH Example 364 OH O CI. HN 0 cl Example 365 o HNS 0 Example 366 o/ HN O
CH
3 WO 2004/080480 PCT/DK2004/000158 198 Example 367 O HN 0 s IN N H Cl Example 368 H N
H
3 C O 0 NH Example 369 0 0 OOH 0 Example 370 O O O H HN I Cl O" O HN aCI 0 Example 371 OO HN Br O Example 372 OO O OOH HIN 0 Example 373 O OlN O ,... N NH HN H 0 WO 2004/080480 PCT/DK2004/000158 199 Example 374 0 S O N H HN 2 0 Example 375 H O N )C S 0, \N / o oz Example 376 o 1. 0N , s HO N 0CH Example 377 0. N o o LCH, Example 378 o
H
3 CO NH H3C O s H 0 Example 379 HN0 0 o _ The following compounds are commercially available and may be prepared using general procedures (B) and / or (C). Example 380 5 5-(5-Bromo-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione OH HN 0 Br WO 2004/080480 PCT/DK2004/000158 200 Example 381 5-Pyridin-4-ylmethylenethiazolidine-2,4-dione 0 HN 0 Example 382 5 5-(3-Bromo-4-methoxybenzylidene)thiazolid ine-2,4-dione HN Br 0 Example 383 5-(3-Nitrobenzylidene)thiazolidine-2,4-dione 0 HNs HNN 10 00 Example 384 5-Cyclohexylidene-1 ,3-thiazolidine-2,4-dione 9>-s HN 0 15 Example 385 5-(3,4-Dihydroxybenzyidene)thiazolidirle-2,4-diofle 0 ts - OH HNI OH 0 20 Example 386 5-(3-Ethoxy-4-hydroxybenzylidene)thiazolidine-2,4-diofle WO 2004/080480 PCT/DK2004/000158 201 0 HN
-
OH C3 Example 387 5-(4-Hyd roxy-3-methoxy-5-nitrobenzylidene)thiazolidine-2,4-dione
H
3
C..
0 HNN 5 0 0 Example 388 5-(3-Ethoxy-4-hyd roxybenzylidene)thiazolidine-2,4-dione CH3 0 0 3 HN !- OH 0 10 Example 389 5-(4-Hydroxy-3,5-dimethoxybenzylidene)thiazolidine-2,4-dione o'C HN s - OH O 0 CH 3 15 Example 390 5-(3-Bromo-5-ethoxy-4-hyd roxybenzyl idene)thiazolid ine-2,4-di!one 0 Br HNS
.
OH 0 KC0 C3 WO 2004/080480 PCT/DK2004/000158 202 Example 391 5-(3-Ethoxy-4-hydroxy-5-nitrobenzylidene)thiazolidine-2,4-dione OO . , N'O o L HNS -S OH OCHCH 5 Example 392 H3 0 0 - o O / HN S 0 CH, s Example 393 H 10 Example 394 H 0 0
N
Example 395 S -o ~~~0
H
3 C O N 15I 15 o WO 2004/080480 PCT/DK2004/000158 203 Example 396 H 5 Example 397 0 NH OHS Example 398 H HN - CH 3 O 10 S s Example 399 o 15 Example 400 5N HN 0 J 0 ° o Gl% WO 2004/080480 PCT/DK2004/000158 204 Example 401 H ~s 0 s F/ 0 5 Example 402 H, Example 403 H3 CH, HO H, NH H z O s== s 10 Example 404 0 H S0 s--( Example 405 15 5-(3-Hydroxy-5-methyl-phenylamino)-thiazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 205
CH
3 HN N OH O H Example 406 iNH S 5 Example 407 H 0 0 S- -- Br Example 408 0 NOO H
CH
3 10 s WO 2004/080480 PCT/DK2004/000158 206 Example 409
).CH
3 O HzC SS N O H Example 410 S NH 5 Ci 0 Example 411 c Ic s ci s ' s H 0 10 Example 412 FF F H Fo-N'O 0 0 WO 2004/080480 PCT/DK2004/000158 207 Example 413 H I0 N H Example 414 0 -OH HN 5S Example 415 OH O HN S S 10 Example 416 0 OH SNH
S
WO 2004/080480 PCT/DK2004/000158 208 Example 417 0 NH S S N CH3 H 3 Example 418 H 0 N Sc Example 419 S S 0 H 0 10 Example 420 H Br S
H
3 C Example 421 H, S 15 N 0 H 15 0 WO 2004/080480 PCT/DK2004/000158 209 Example 422 0 H N S Br N H Example 423 0CH N S S>S NO s ~s 5 0 Example 424 I I 150 H Cl- O S 10 Example 425
CH
3 HC N O S NH S Example 426 H 0 B N Br S" 15 3 WO 2004/080480 PCT/DK2004/000158 210 Example 427 H3C N 3 S N'.CHz H 0 N / Example 428
H
0 N 5 Example 429
H
3 C 0 HzNH *N 0 Example 430 0 10 HN KS Example 431 5-(4-Diethylamino-2-methoxy-benzylidene)-imidazolidine-2,4-dione WO 2004/080480 PCT/DK2004/000158 211 HO N HaC-.
0
H
3 c7C N 0 N
H
3 C Example 432 o 0 HN N CH3 5 Example 433
CH
3 0 OHN 3 H0 Example 434 H3c
H
3 C 0 O N " NH N S 10 0 Example 435 WO 2004/080480 PCT/DK2004/000158 212 HzC' H3C-.
0
H
3 C N O H'N H3 C Example 436 HO I=NH HO 0 N O H 5 Example 437 H ON OO o /s o o Example 438 H N "S -j I 10 m 4CH3 10 H3 CO Example 439 WO 2004/080480 PCT/DK2004/000158 213
CH
3 HO N N S
CH
3 Example 440 I NH S s 5 Example 441 H S NO s CI - 0 Example 442 0 S N10 \ -CH3 10 WO 2004/080480 PCT/DK2004/000158 214 Example 443
-
F HN F F 0 Example 444
CH
3 HN
CH
3 5 Example 445 H Cl N N HO 0 H 0
CH
3 10 Example 446 0 HN SF Example 447 Example 447 WO 2004/080480 PCT/DK2004/000158 215 CH3 O) />NH2 Example 448 0 HN o S NH 0 5 Example 449 0 N S NH H NH Example 450 H N0 S=-< HN 0 10 Example 451 WO 2004/080480 PCT/DK2004/000158 216 H O N S HO 0 Example 452 0 HO' N/ 0 5 Example 453 O H 00/ N 0 Example 454 10 5-(4-Diethylamino-benzylidene)-2-imino-thiazolidin-4-one
H
3 C) N CH 3 o 0s NH Example 455 0 o H N O S
CH
3 WO 2004/080480 PCT/DK2004/000158 217 Example 456 O HN HN / 5 Example 457 O H N s >= S 0 ci CI Example 458 CH 0 0 lOO 10 Example 459 H H S
N"S
WO 2004/080480 PCT/DK2004/000158 218 General procedure (D) for preparation of compounds of general formula 13: 0 A, OH + Step 1 O A O'R' Step 2 OC A OH "O Lea/ O<AO/ Y~R ~A 0 O R O R O R O Step 3 Y (CH 2 )n HN AO OH O R 3 O 13 5 wherein X, Y, and R 3 are as defined above, n is 1 or 3-20, E is arylene or heterarylene (including up to four optional substituents, R 13 , R 14 , R 1 5 , and R 1 5 A as defined above), 10 R' is a standard carboxylic acid protecting group, such as Cl-C 6 -alkyl or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the like. Step 1 is an alkylation of a phenol moiety. The reaction is preformed by reacting Rio-C(=O) 15 E-OH with an w-bromo-alkane-carboxylic acid ester (or a synthetic equivalent) in the pres ence of a base such as sodium or potassium carbonate, sodium or potassium hydroxide, so dium hydride, sodium or potassium alkoxide in a solvent, such as DMF, NMP, DMSO, ace tone, acetonitrile, ethyl acetate or isopropyl acetate. The reaction is performed at 20 - 160 oC, usually at room temperature, but when the phenol moiety has one or more substituents 20 heating to 50 OC or more can be beneficial, especially when the substituents are in the ortho position relatively to the phenol. This will readily be recognised by those skilled in the art. Step 2 is a hydrolysis of the product from step 1.
WO 2004/080480 PCT/DK2004/000158 219 Step 3 is similar to general procedure (B) and (C). This general procedure (D) is further illustrated in the following examples: 5 Example 460 (General procedure (D)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid O 0 0 OO HN 0 Step 1: A mixture of 4-hydroxybenzaldehyde (9.21 g, 75 mmol), potassium carbonate (56 g, 410 10 mmol) and 4-bromobutyric acid ethyl ester (12.9 mL, 90 mmol) in N,N-dimethylformamide (250 mL) was stirred vigorously for 16 hours at room temperature. The mixture was filtered and concentrated in vacuo to afford 19.6 g (100%) of 4-(4-formylphenoxy)butyric acid ethyl ester as an oil. 1 H-NMR (DMSO-d 6 ): 8 1.21 (3H, t), 2.05 (2H, p), 2.49 (2H, t), 4.12 (4H, mn), 7.13 (2H, d), 7.87 (2H, d), 9.90 (1 H, s). HPLC-MS (Method A): m/z = 237 (M+1); Rt = 3.46 15 min. Step 2: 4-(4-Formylphenoxy)butyric acid ethyl ester (19.6 g, 75 mmol) was dissolved in methanol (250 mL) and 1N sodium hydroxide (100 mL) was added and the resulting mixture was 20 stirred at room temperature for 16 hours. The organic solvent was evaporated in vacuo (40 OC, 120 mBar) and the residue was acidified with 1N hydrochloric acid (110 mL). The mixture was filtered and washed with water and dried in vacuo to afford 14.3 g (91%) 4-(4 formylphenoxy)butyric acid as a solid. 1 H-NMR (DMSO-d 6 ): 5 1.99 (2H, p), 2.42 (2H, t), 4.13 (2H, t), 7.14 (2H, d), 7.88 (2H, d), 9.90 (1H, s), 12.2 (1H, bs). HPLC-MS (Method A): m/z = 25 209 (M+1); Rt = 2.19 min. Step 3: Thiazolidine-2,4-dione (3.55 g, 27.6 mmol), 4-(4-formylphenoxy)butyric acid (5.74 g, 27.6 mmol), anhydrous sodium acetate (11.3 g, 138 mmol) and acetic acid (100 mL) was refluxed 30 for 16 h. After cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 2.74 g (32%) of 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid as a solid.
WO 2004/080480 PCT/DK2004/000158 220 1 H-NMR (DMSO-de): t1.97 (2H, p), 2.40 (2H, t), 4.07 (2H, t), 7.08 (2H, d), 7.56 (2H, d), 7.77 (1H, s), 12.2 (1H, bs), 12.5 (1H, bs); HPLC-MS (Method A): m/z: 308 (M+1); Rt = 2.89 min. Example 461 (General procedure (D)) 5 [3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid 0 HN H OY OH O 0 Step 3: Thiazolidine-2,4-dione (3.9 g, 33 mmol), 3-formylphenoxyacetic acid (6.0 g, 33 mmol), anhy drous sodium acetate (13.6 g, 165 mmol) and acetic acid (100 mL) was refluxed for 16 h. Af 10 ter cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 5.13 g (56%) of [3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid as a solid. 1 H-NMR (DMSO-d 6 ): 6 4.69 (2H, s), 6.95 (1H, dd), 7.09 (1H, t), 7.15 (1H, d), 7.39 (1H, t),7.53 (1 H, s); HPLC-MS (Method A): mlz = 280 (M+1) (poor ionisation); Rt = 2.49 min. 15 The compounds in the following examples were similarly prepared. Example 462 (General procedure (D)) 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acrylic acid 0 O O S OH HN 20 O 1 H-NMR (DMSO-d 6 ): 86.63 (1H, d), 7.59-7.64 (3H, m), 7.77 (1H, s), 7.83 (2H, m). Example 463 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid WO 2004/080480 PCT/DK2004/000158 221 0 0 H - O"1 OH HN I O Triethylamine salt: 1 H-NMR (DMSO-d 6 ): 3 4.27 (2H, s), 6.90 (2H, d), 7.26 (1H, s), 7.40 (2H, d). 5 Example 464 (General procedure (D)) 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoic acid UO S OH HN O 10 Example 465 (General procedure (D)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoic acid HN OH 0 0 1 H-NMR (DMSO-d 6 ): 6 7.57 (1H, s), 7.60 (1H, t), 7.79 (1H, dt), 7.92 (1H, dt), 8.14 (1H, t). 15 Example 466 (General procedure (D)) 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid 0 O HN Cl 0 1 H-NMR (DMSO-d 6 ): 8 2.00 (2H, p), 2.45 (2H, t), 4.17 (2H, t), 7.31 (1H, d), 7.54 (1H, dd), 7.69 (1H, d), 7.74 (1H, s), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 364 20 (M+23); Rt = 3.19 min. Example 467 (General procedure (D)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid WO 2004/080480 PCT/DK2004/000158 222 0 0 HNO HN rO Br 0 1 H-NMR (DMSO-d 6 ): 8 1.99 (2H, p), 2.46 (2H, t), 4.17 (2H, t), 7.28 (1H, d), 7.57 (1H, dd), 7.25 (1H, s), 7.85 (1H, d), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3.35 min. 5 Example 468 (General procedure (D)) 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid S 0 HN 0 OH HNBr O 0 1 H-NMR (DMSO-d 6 ): 5 1.99 (2H, p), 2.45 (2H, t), 4.18 (2H, t), 7.28 (1H, d), 7.55 (1H, dd), 10 7.60 (1H, s), 7.86 (1H, d), 12.2 (1 H, bs), 13.8 (1 H, bs). HPLC-MS (Method A): m/z: 424 (M+23); Rt = 3.84 min. HPLC-MS (Method A): mlz: 424 (M+23); Rt = 3,84 min Example 469 (General procedure (D)) 15 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid 00 HN O 0 1 H-NMR (DMSO-d): 3 2.12 (2H, p), 2.5 (below DMSO), 4.28 (2H, t), 7.12 (1H, d), 7.6-7.7 (3H, m), 8.12 (1H, d), 8.31 (1H, d), 8.39 (1H, s), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 380 (M+23); Rt = 3.76 min. 20 Example 470 (General procedure (D)) 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoic acid WO 2004/080480 PCT/DK2004/000158 223 O S O1 0 , , , ,O HN OH O HPLC-MS (Method A): m/z: 394 (M+23); Rt = 3.62 min. 1 H-NMR (DMSO-d 6 ): 8 1.78 (2H, m), 1.90 (2H, m), 2.38 (2H, t), 4.27 (2H, t), 7.16 (1H, d), 7.6-7.75 (3H, m), 8.13 (1H, d), 8.28 (1H, d), 8.39 (1H, s), 12.1 (1H, bs), 12.6 (1H, bs). 5 Example 471 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoic acid. HN N .Br OH O 0 10 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]pentanoic acid (example 470, 185 mg, 0.5 mmol) was treated with an equimolar amount of bromine in acetic acid (10 mL). Stirring at RT for 14 days followed by evaporation to dryness afforded a mixture of the bro minated compound and unchanged starting material. Purification by preparative HPLC on a C18 column using acetonitrile and water as eluent afforded 8 mg of the title compound. 15 HPLC-MS (Method C): mlz: 473 (M+23), Rt. = 3.77 min Example 472 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid. 0 0 ,- - O O O OH HN& Br 20 0 Starting with 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyric acid (ex ample 469, 0.5 mmol) using the same method as in example 471 afforded 66 mg of the title compound. 25 HPLC-MS (Method C): m/z: 459 (M+23); Rt. = 3.59 min.
WO 2004/080480 PCT/DK2004/000158 224 Example 473 (General procedure (D)) [2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid O HNO O 5 1 H-NMR (DMSO-d 6 ): 8 4.90 (2H, s), 7.12 (1H, d), 7.52 (1H, dd), 7.65 (1H, s) 7.84 (1H, d).HPLC-MS (Method A): m/z: not observed; Rt = 2.89 min. Example 474 (General procedure (D)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid 0 HN OH 10 o o 1 H-NMR (DMSO-d 6 ): 8 1.98 (2H, p), 2.42 (2H, t), 4.04 (2H, t), 7.05 (1H, dd), 7.15 (2H, m), 7.45 (1H, t), 7.77 (1H, s), 12.1 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 330 (M+23); Rt = 3.05 min. 15 Example 475 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-3-methoxyphenoxy]acetic acid 00 H No OH O
OCH
3 HPLC-MS (Method B): m/z: 310 (M+1); Rt = 3,43 min. 20 Example 476 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetic acid WO 2004/080480 PCT/DK2004/000158 225 O O OS- O OH HN O HPLC-MS (Method A): m/z: 330 (M+1); Rt = 3.25 min. Example 477 (General procedure (D)) 5 8-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1l-carboxylic acid O S/ HN O HO O HPLC-MS (Method A): m/z: 299 (M+1); Rt = 2,49 min. Example 478 (General procedure (D)) 10 [3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1 -yl]acetic acid O >_N OH HN 0 HPLC-MS (Method A): m/z: 303 (M+1); Rt = 2.90 min. Preparation of starting material: 15 3-Formylindol (10 g, 69 mmol) was dissolved in N,N-dimethylformamide (100 mL) and under an atmosphere of nitrogenand with external cooling, keeping the temperature below 15 oC, sodium hydride (60% in mineral oil, 3.0 g, 76 mmol) was added in portions. Then a solution of ethyl bromoacetate (8.4 mL, 76 mmol) in N,N-dimethylformamide (15 mL) was added dropwise over 30 minutes and the resulting mixture was stirred at room temperature for 16 20 hours. The mixture was concentrated in vacuo and the residue was partitioned between wa ter (300 mL) and ethyl acetate (2 x 150 mL). The combined organic extracts were washed with a saturated aqueous solution of ammonium chloride (100 mL), dried (MgSO 4 ) and con- WO 2004/080480 PCT/DK2004/000158 226 centrated in vacuo to afford 15.9 g (quant.) of (3-formylindol-1-yl)acetic acid ethyl ester as an oil. 1 H-NMR (CDCI 3 ): 6 H = 1.30 (3H, t), 4.23 (2H, q), 4.90 (2H, s), 7.3 (3H, m), 7.77 (1H, s), 8.32 5 (1H, d), 10.0 (1H, s). (3-Formylindol-1 -yl)acetic acid ethyl ester (15.9 g 69 mmol) was dissolved in 1,4-dioxane (100 mL) and 1N sodium hydroxide (10 mL) was added and the resulting mixture was stirred at room temperature for 4 days. Water (500 mL) was added and the mixture was washed 10 with diethyl ether (150 mL). The aqueous phase was acidified with 5N hydrochloric acid and extracted with ethyl acetate (250 + 150 mL). The combined organic extracts were dried (MgSO 4 ) and concentrated in vacuo to afford 10.3 g (73%) of (3-formylindol-1-yl)acetic acid as a solid. 15 1 H-NMR (DMSO-de): 6 H = 5.20 (2H, s), 7.3 (2H, m), 7.55 (1H, d), 8.12 (1H, d), 8.30 (1H, s), 9.95 (1H, s), 13.3 (1H, bs). Example 479 (General procedure (D)) 3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1 -yl]propionic acid 0 OH HN 20 O HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.08 min. Preparation of starting material: A mixture of 3-formylindol (10 g, 69 mmol), ethyl 3-bromopropionate (10.5 mL, 83 mmol) and 25 potassium carbonate (28.5 g, 207 mmol) and acetonitrile (100 mL) was stirred vigorously at refux temperature for 2 days. After cooling, the mixture was filtered and the filtrate was con centrated in vacuo to afford 17.5 g (quant.) of 3-(3-formylindol-1-yl)propionic acid ethyl ester as a solid.
WO 2004/080480 PCT/DK2004/000158 227 1 H-NMR (DMSO-d 6 ): 6 H = 1.10 (3H, t), 2.94 (2H, t), 4.02 (2H, q), 4.55 (2H, t), 7.3 (2H, m), 7.67 (1H, d), 8.12 (1H, d), 8.30 (1H, s), 9.90 (1H, s). 3-(3-Formylindol-1-yl)propionic acid ethyl ester (17.5 g 69 mmol) was hydrolysed as de 5 scribed above to afford 12.5 g (83%) of 3-(3-formylindol-1-yl)propionic acid as a solid. 1 H-NMR (DMSO-d 6 ): 6 H = 2.87 (2H, t), 4.50 (2H, t), 7.3 (2H, m), 7.68 (1H, d), 8.12 (1H, d), 8.31 (1H, s), 9.95 (1H, s), 12.5 (1H, bs). 10 Example 480 (General procedure (D)) {5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3-yl}acetic acid 0 OH S 0 0 O 0 HPLC-MS (Method A): m/z: 429 (M+23); Rt = 3.89 min. 15 Example 481 (General procedure (D)) 6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxyoctanoic acid 0 0 OH HN 0 20 HPLC-MS (Method C): m/z: 436 (M+23); Rt.= 4.36 min The intermediate aldehyde for this compound was prepared by a slightly modified procedure: 6-Hydroxynaphthalene-2-carbaldehyde (1.0 g, 5.8 mmol) was dissolved in DMF (10 mL) and sodium hydride 60% (278 mg) was added and the mixture stirred at RT for 15 min. 8 25 Bromooctanoic acid (0.37 g, 1.7 mmol) was converted to the sodium salt by addition of so dium hydride 60% and added to an aliquot (2.5 mL) of the above naphtholate solution and the resulting mixture was stirred at RT for 16 hours. Aqueous acetic acid (10 %) was added and the mixture was extracted 3 times with diethyl ether. The combined organic phases were WO 2004/080480 PCT/DK2004/000158 228 dried with MgSO 4 and evaporated to dryness affording 300 mg of 8-(6-formylnaphthalen-2 yloxy)octanoic acid. HPLC-MS (Method C): m/z 315 (M+1); Rt. = 4.24 min. 5 Example 482 (General procedure (D)) 12-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]dodecanoic acid. O O O OH HN I 0 HPLC-MS (Method C): m/z: 492 (M+23); Rt.= 5.3 min. 10 The intermediate aldehyde was prepared similarly as described in example 481. Example 483 (General procedure (D)) 11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoic acid. 0 NHNO OH HN 15 HPLC-MS (Method C): m/z:478 (M+23); Rt.= 5.17 min. The intermediate aldehyde was prepared similarly as described in example 481. 20 Example 484 (General procedure (D)) 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoic acid. 0 O 0 OH HIN 0 0 HPLC-MS (Method C): m/z: 534 (M+23); Rt.= 6.07 min. 25 The intermediate aldehyde was prepared similarly as described in example 481. Example 485 (General procedure (D)) 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoic acid.
WO 2004/080480 PCT/DK2004/000158 229 o o H HN I o HPLC-MS (Method C): m/z: 408 (M+23); Rt.= 3.71 min. Example 486 (General procedure (D)) 5 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyric acid. H O,, OH o HN I O 0 HPLC-MS (Method C): m/z: 380 (M+23); Rt.= 3.23 min. Example 487 (General procedure (D)) 10 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoic acid ethyl ester. HN HN I. o.
o 0 HPLC-MS (Method C): m/z: 436 (M+23); Rt.= 4.64 min. Example 488 (General procedure (D)) 15 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyric acid ethyl ester. 0 HN I O N OCH 3 0 HPLC-MS (Method C): m/z: 408 (M+23); Rt.= 4.28 min. Example 489 (General procedure (D)) 20 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentyl}malonic acid WO 2004/080480 PCT/DK2004/000158 230 ) =0 S HO 0 O H HPLC-MS (Method C): m/z = 444 (M+1); Rt = 3,84 min. Example 490 (General procedure (D) 5 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentyl}malonic acid diethyl ester H O N S 0
H
3 C O 0O O ON CH, HPLC-MS (Method C): m/z = 500 (M+1); Rt = 5.18 min. 10 Example 491 (General procedure (D)) 4-[4-(2,4,6-Trioxotetrahydropyrimidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid H O O N 0 0 0 H HN HPLC-MS (Method C): m/z = 369 (M+1); Rt = 2,68 min. 15 Example 492 N-(3-Aminopropyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-1 -yloxy] butyramide WO 2004/080480 PCT/DK2004/000158 231 o s HN H 0 To a mixture of 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid (example 469, 5.9 g, 16.5 mmol) and 1-hydroxybenzotriazole (3.35 g, 24.8 mmol) in DMF (60 5 mL) was added 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (4.75 g, 24.8 mmol) and the resulting mixture was stirred at room temperature for 2 hours. N-(3-amino propylcarbamic acid tert-butyl ester (3.45 g, 19.8 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and ethyl acetate and dichloromethane were added to the residue. The mixture was filtered, 10 washed with water and dried in vacuo to afford 4.98 g (59%) of (3-{4-[4-(2,4-dioxothiazolidin 5-ylidenemethyl)naphthalen-1-yloxy]butyrylamino}propyl)carbamic acid tert-butyl ester. HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.79 min. 15 (3-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyrylamino} propyl)carbamic acid tert-butyl ester (4.9 g, 9.5 mmol) was added dichloromethane (50 mL) and trifluoroacetic acid (50 mL) and the resulting mixture was stirred at room temperature for 45 minutes. The mixture was concentrated in vacuo and co-evaporated with toluene. To the residue was added ethyl acetate (100 mL) and the mixture was filtered and dried in vacuo to 20 afford the title compound as the trifluoroacetic acid salt. HPLC-MS (Method C): m/z: 414 (M+1); Rt = 2,27 min. Compounds of the invention includes: 25 WO 2004/080480 PCT/DK2004/000158 232 Example 493 0 0 0 HNO 0 Example 494 HN O0 OH HN / 0 O Example 495 o 0 0 °[(°N- OH HN 0 Example 496 0 H HN 0 0 Example 497 OO 0 OH HN N 0 Example 498 HN 0 O O 0 Example 499 WO 2004/080480 PCT/DK2004/000158 233 o
-
OH HN 0 Example 500 S ON OH 0 1 0 HN .O 0 Example 501 00 OO O-S OOH OO HN N 0 Example 502 HN0 O O 0 Example 503 OO S 0OH O-S O O HN 0 Example 504 (Prepared analogously to General Procedure (D)) 5 2-{5-[4-(2,4-Thiazolidindion-5-ylidenemethyl)naphthalen-1 -yloxy]pentyl}malonic acid WO 2004/080480 PCT/DK2004/000158 234 0 HN OS/ 0/ o 0 OH HO O0 A solution of 4-hydroxy-1l-naphtaldehyde (1.0 g, 5.81 mmol), 2-(5-bromopentyl)malonic acid diethyl ester (2.07 g, 6.68 mmol) and potassium carbonate (4.01 g, 29 mmol) in DMF (50 mL) was stirred at 1000 C for 3 hours. The mixture was cooled and the salt was filtered off. The 5 solvent was then removed under reduced pressure to afford 2.9 g of crude 2-[5-(4 formylnaphtalen-1-yloxy)pentyl]malonic acid diethyl ester which was used for the next reac tion without further purification. HPLC-MS (Method C): m/z: 401 (M+1); Rt= 5.16 min. 1 H-NMR (DMSO-d6): 65 = 1.18 (t, 6 H), 10 1,39 (m, 2 H), 1.55 (m, 2 H), 1.87 (m, 4 H), 3.48 (t, 1 H), 4.13 (m, 4 H), 4.27 (t, 2 H), 7.17 (d, 1 H), 7.64(t, 1 H), 7.75 (t, 1 H), 8.13 (d, 1 H), 8.29 (d, 1 H), 9.24 (d, 1 H), 10.19 (s, 1 H). 1.4 g (3.5 mmol) of crude 2-[5-(4-formylnaphtalen-1-yloxy)pentyl]malonic acid diethyl ester was treated with aqueous sodium hydroxide (1N, 8.75 mL, 8.75 mmol) and methanol (50 15 mL). The solution was stirred at 700 C for 5 hours and the mixture was concentrated under reduced pressure. Hydrochloric acid (6 N) was added until pH <2. The resulting slurry was stirred untill it solidified. The crystals were filtered off, washed with water and then dried in vacuo to afford 1.1 g (92%) of 2-[5-(4-formylnaphtalen-1-yloxy)pentyl]malonic acid. The product was used in the next step without further purification. 20 HPLC-MS (Method C): m/z: 345 (M+1); Rt = 3.52 min. 1 H-NMR(DMSO-d6): 6 = 1,40 (m, 2 H), 1.55 (m, 2 H), 1.80 (m, 2 H), 1.90 (m, 2 H), 3.24 (t, 1 H), 4.29 (t, 2 H), 7.19 (d, 1 H), 7.64(t, 1 H), 7.75 (t, 1 H), 8.14 (d, 1 H), 8.30 (d, 1 H), 9.23 (d, 1 H), 10.18 (s, 1 H), 12.69 (s, 2 H). 25 To a solution of 2-[5-(4-formylnaphtalen-1 -yloxy) pentyl]malonic acid (0.36 g, 1.05 mmol) in acetic acid (10 mL) was added 2,4-thiazolidindione (0.16 g,1.36 mmol) and piperidine (0.52 mL, 5.25 mmol). The solution was heated to 105 OC for 24 hours. After cooling to room tem perature, the solvents were removed in vacuo. Water was added to the residue. The precipi- WO 2004/080480 PCT/DK2004/000158 235 tate was filtered off and washed with water. Recrystalisation from acetonitrile afforded 200 mg (43%) of the title compound as a solid. HPLC-MS (Method C): m/z: 422 (M-CO 2 +Na); Rt = 4.08 min. 1 H-NMR(DMSO-d 6 ): 6 = 1,41 5 (m, 2 H), 1.55 (m, 4 H), 1.88 (m, 2 H), 2.23 (t, 1 H), 4.24 (t, 2 H), 7.61-7.74 (m, 3 H), 8.12 (d, 1 H), 8.28 (d, 1 H), 8.38 (s, 1 H), 12.00 (s, 1 H), 12.59 (s, 2 H). 10 The following compounds are commercially available and may be prepared according to general procedure (D): Example 505 OH 0 o HN 0a O CH, 15 Example 506 O OH Example 507 S O OH 20 WO 2004/080480 PCT/DK2004/000158 236 Example 508 0 0 HzC S H HO S HO 0 Example 509 0 O O / O OH HN .S O-CH 3 5 S Example 510 0 SO HN O N 0 / HO 10 Example 511 H S O OH 0 0 15 The following salicylic acid derivatives do all bind to the His B10 Zn2+ site of the insulin hexamer: WO 2004/080480 PCT/DK2004/000158 237 Example 512 Salicylic acid O HO HO 5 Example 513 Thiosalicylic acid (or: 2-Mercaptobenzoic acid) 0 O HS Example 514 10 2-Hydroxy-5-nitrobenzoic acid OH O 11+ 0 " N, O 07 o HO Example 515 3-Nitrosalicyclic acid O HO HO 15 0 Example 516 5,5'-Methylenedisalicylic acid O OH O OH HO OH I I WO 2004/080480 PCT/DK2004/000158 238 Example 517 2 -Amino-5-trfluoromethylbenzoesyre OH F 0 F
H
2 N 5 Example 518 2 -Amino-4-chlorobenzoic acid 0 O HO -1
H
2 N Cl 10 Example 519 2 -Amino-5-methoxybenzoesyre OH O O, CH 3
H
2 N Example 520 15 0 HO CI
H
2 N Example 521 0 HO Br
H
2 N 20 WO 2004/080480 PCT/DK2004/000158 239 Example 522 OH O==O HO
NH
2 Example 523 OH
H
2 Cl Example 524 0 H O 0 ,o 10 Example 525 O HO H 2 N1 OCH Example 526 5-lodosalicylic acid O HO 15 HO Example 527 5-Chlorosalicylic acid WO 2004/080480 PCT/DK2004/000158 240 O HO C HO Example 528 1-Hydroxy-2-naphthoic acid OH OH o 5 Example 529 3,5-Dihydroxy-2-naphthoic acid 0 HO HO -/ HO OH 10 Example 530 3-Hydroxy-2-naphthoic acid O HO HO 15 Example 531 3,7-Dihydroxy-2-naphthoic acid 0 HO HOH O H Example 532 20 2-Hydroxybenzo[a]carbazole-3-carboxylic acid WO 2004/080480 PCT/DK2004/000158 241 0 O HO -/ HO N
H
Example 533 7 -Bromo-3-hydroxy-2-naphthoic acid 0 HO Br HO 5
HOHO
B r This compound was prepared according to Murphy et al., J. Med. Chem. 1990, 33,171-8. HPLC-MS (Method A): m/z: 267 (M+1); Rt: = 3.78 min. Example 534 10 1,6-Dibromo-2-hydroxynaphthalene-3-carboxylic acid O Br HO Br HO Br This compound was prepared according to Murphy et al, J. Med. Chem. 1990, 33, 171-8. HPLC-MS (Method A): m/z: 346 (M+1); Rt: = 4,19 min. 15 Example 535 7-Formyl-3-hyd roxynaphthalene-2-carboxylic Acid O 0 HOH ~ H IHO A solution of 7-bromo-3-hydroxynaphthalene-2-carboxylic acid (15.0 g, 56.2 mmol) (example 533) in tetrahydrofuran (100 mL) was added to a solution of lithium hydride (893 mg, 112 20 mmol) in tetrahydrofuran (350 mL). After 30 minutes stirring at room temperature, the result ing solution was heated to 50 OC for 2 minutes and then allowed to cool to ambient tempera ture over a period of 30 minutes. The mixture was cooled to -78 OC, and butyllithium (1.6 M in hexanes, 53 mL, 85 mmol) was added over a period of 15 minutes. N,N-Dimethylformamide WO 2004/080480 PCT/DK2004/000158 242 (8.7 mL, 8.2 g, 112 mmol) was added after 90 minutes additional stirring. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 17 hours before it was poured into 1 N hydrochloric acid (aq.) (750 mL). The organic solvents were evaporated in vacuo, and the resulting precipitate was filtered off and rinsed with water (3 x 100 mL) to 5 yield the crude product (16.2 g). Purification on silica gel (dichloromethane / methanol / ace tic acid = 90:9:1) furnished the title compound as a solid. 1 H-NMR (DMSO-d 6 ): 811.95 (1H, bs), 10.02 (1H, s), 8.61 (1H, s), 8.54 (1H, s), 7.80 (2H, bs), 7.24 (1 H, s); HPLC-MS (Method (A)): m/z: 217 (M+1); Rt = 2.49 min. 10 Example 536 3-Hydroxy-7-methoxy-2-naphthoic acid 0 HO O~ 'C H HO 15 Example 537 4-Amino-2-hydroxybenzoic acid O HO HO
NH
2 Example 538 20 5-Acetylamino-2-hydroxybenzoic acid 0 H HOCH 0 Example 539 2-Hydroxy-5-methoxybenzoic acid WO 2004/080480 PCT/DK2004/000158 243 O CH 3 HOO HO HO The following compounds were prepared as described below: Example 540 5 4-Bromo-3-hydroxynaphthalene-2-carboxylic acid O HO HO Br 3-Hydroxynaphthalene-2-carboxylic acid (3.0 g, 15.9 mmol) was suspended in acetic acid (40 mL) and with vigorous stirring a solution of bromine (817 pL, 15.9 mmol) in acetic acid (10 mL) was added drop wise during 30 minutes. The suspension was stirred at room tern 10 perature for 1 hour, filtered and washed with water. Drying in vacuo afforded 3.74 g (88%) of 4-bromo-3-hydroxynaphthalene-2-carboxylic acid as a solid. 1 H-NMR (DMSO-d 6 ): 8 7.49 (1H, t), 7.75 (1H, t), 8.07 (2H, "t"), 8.64 (1H, s). The substitution pattern was confirmed by a COSY experiment, showing connectivities between the 3 (4 hy drogen) "triplets". HPLC-MS (Method A): mlz: 267 (M+1); Rt = 3.73 min. 15 Example 541 3-Hydroxy-4-iodonaphthalene-2-carboxylic acid O HO HO I 3-Hydroxynaphthalene-2-carboxylic acid (0.5 g, 2.7 mmol) was suspended in acetic acid (5 20 mL) and with stirring iodine monochloride (135 gL, 2.7 mini) was added. The suspension was stirred at room temperature for 1 hour, filtered and washed with water. Drying afforded 0.72 g (85%) of 4-iodo-3-hydroxynaphthalene-2-carboxylic acid as a solid. 1 H-NMR (DMSO-d 6 ): 8 7.47 (1H, t), 7.73 (1H, t), 7.98 (1H, d), 8.05 (1H, d), 8.66 (1H, s). HPLC-MS (Method A): m/z: 315 (M+1); Rt = 3.94 min.
WO 2004/080480 PCT/DK2004/000158 244 Example 542 2-Hydroxy-5-[(4-methoxyphenylamino)methyl]benzoic acid CH3 00 HO HO N 5 p-Anisidine (1.3 g, 10.6 mmol) was dissolved in methanol (20 mL) and 5-formylsalicylic acid (1.75 g, 10.6 mmol)was added and the resulting mixture was stirred at room temperature for 16 hours. The solid formed was isolated by filtration, re-dissolved in N-methyl pyrrolidone (20 mL) and methanol (2 mL). To the mixture was added sodium cyanoborohydride (1.2 g) and 10 the mixture was heated to 70 oC for 3 hours. To the cooled mixture was added ethyl acetate (100 mL) and the mixture was extracted with water (100 mL) and saturated aqueous ammo nium chloride (100 mL). The combined aqueous phases were concentrated in vacuo and a 2 g aliquot was purified by SepPac chromatography eluting with mixtures of aetonitrile and wa ter containing 0.1% trifluoroacetic acid to afford the title compound. 15 HPLC-MS (Method A): m/z: 274 (M+1); Rt = 1.77 min. 1 H-NMR (methanol-d 4 ): 8 3.82 (3H, s), 4.45 (2H, s), 6.96 (1 H, d), 7.03 (2H, d), 7.23 (2H, d), 7.45 (1 H, dd), 7.92 (1H, d). 20 Example 543 2-Hydroxy-5-(4-methoxyphenylsulfamoyl)benzoic acid
CH
3 HO , N HH HO A solution of 5-chlrosulfonylsalicylic acid (0.96 g, 4.1 mmol) in dichloromethane (20 mL) and 25 triethylamine (1.69 mL, 12.2 mmol) was added p-anisidine (0.49 g, 4.1 mmol) and the result ing mixture was stirred at room temperature for 16 hours. The mixture was added dichloro methane (50 mL) and was washed with water (2 x 100 mL). Drying (MgSO 4 ) of the organic phase and concentration in vacuo afforded 0.57 g crude product. Purification by column WO 2004/080480 PCT/DK2004/000158 245 chromatography on silica gel eluting first with ethyl acetate:heptane (1:1) then with methanol afforded 0.1 g of the title compound. HPLC-MS (Method A): m/z: 346 (M+23); Rt = 2.89 min. 5 1 H-NMR (DMSO-d 6 ): 6 3.67 (3H, s), 6.62 (1H, d), 6.77 (2H, d), 6.96 (2H, d), 7.40 (1H, dd), 8.05 (1H, d), 9.6 (1H, bs). General procedure (E) for preparation of compounds of general formula 14: 0 Lea s Pd catalyst 0 HO 1+ .B -j Base 3 H J H HO 0 R H R HOI 10 14 wherein Lea is a leaving group such as Cl, Br, I or OSO 2
CF
3 , R is hydrogen or Cl-C 6 -alkyl, optionally the two R-groups may together form a 5-8 membered ring, a cyclic boronic acid ester, and J is as defined above. 15 An analogous chemical transformation has previously been described in the literature (Bumagin et al., Tetrahedron, 1997, 53, 14437-14450). The reaction is generally known as the Suzuki coupling reaction and is generally performed by reacting an aryl halide or triflate with an arylboronic acid or a heteroarylboronic acid in the presence of a palladium catalyst 20 and a base such as sodium acetate, sodium carbonate or sodium hydroxide. The solvent can be water, acetone, DMF, NMP, HMPA, methanol, ethanol toluene or a mixture of two or more of these solvents. The reaction is performed at room temperature or at elevated temperature. The general procedure (E) is further illustrated in the following example: 25 Example 544 (General Procedure (E)) 7-(4-Acetylphenyl)-3-hydroxynaphthalene-2-carboxylic Acid 0 0 CH HO
HO
WO 2004/080480 PCT/DK2004/000158 246 To 7-bromo-3-hydroxynaphthalene-2-carboxylic acid (100 mg, 0.37 mmol) (example 533) was added a solution of 4-acetylphenylboronic acid (92 mg, 0.56 mmol) in acetone (2.2 mL) followed by a solution of sodium carbonate (198 mg, 1.87 mmol) in water (3.3 mL). A sus pension of palladium(ll) acetate (4 mg, 0.02 mmol) in acetone (0.5 mL) was filtered and 5 added to the above solution. The mixture was purged with N 2 and stirred vigorously for 24 hours at room temperature. The reaction mixture was poured into 1 N hydrochloric acid (aq.) (60 mL) and the precipitate was filtered off and rinsed with water (3 x 40 mL). The crude product was dissolved in acetone (25 mL) and dried with magnesium sulfate (1 h). Filtration followed by concentration furnished the title compound as a solid (92 mg). 10 1 H-NMR (DMSO-d): 812.60 (1 H, bs), 8.64 (1H, s), 8.42 (1H, s), 8.08 (2H, d), 7.97 (2H, d), 7.92 (2H, m), 7.33 (1H, s), 2.63 (3H, s); HPLC-MS (Method (A): m/z: 307 (M+1); Rt = 3.84 min. The compounds in the following examples were prepared in a similar fashion. Optionally, the 15 compounds can be further purified by recrystallization from e.g. ethanol or by chromatogra phy. Example 545 (General Procedure (E)) 3-Hydroxy-7-(3-methoxyphenyl)naphthalene-2-carboxylic acid 0 HO 0 20 HOCH 3 HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.60 min. Example 546 (General Procedure (E)) 3-Hydroxy-7-phenylnaphthalene-2-carboxylic acid 0 HO 25 HO HPLC-MS (Method (A)): m/z: 265 (M+1); Rt = 4.6 min. Example 547 (General Procedure (E)) 3-Hydroxy-7-p-tolylnaphthalene-2-carboxylic acid WO 2004/080480 PCT/DK2004/000158 247
CH
3 0 NI HO HO HPLC-MS (Method (A)): m/z: 279 (M+1); Rt = 4.95 min. Example 548 (General Procedure (E)) 5 7-(4-Formylphenyl)-3-hydroxynaphthalene-2-carboxylic acid O O H HO HPLC-MS (Method (A)): m/z: 293 (M+1); Rt = 4.4 min. Example 549 (General Procedure (E)) 10 6-Hydroxy-[1,2]binaphthalenyl-7-carboxylic acid o HO HO HPLC-MS (Method (A)): m/z: 315 (M+1); Rt = 5.17 min. Example 550 (General Procedure (E)) 15 7-(4-Carboxy-phenyl)-3-hydroxynaphthalene-2-carboxylic acid O 0 / OH HO HO HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 3.60 min. Example 551 (General Procedure (E)) 20 7-Benzofuran-2-yl-3-hydroxynaphthalene-2-carboxylic acid WO 2004/080480 PCT/DK2004/000158 248 O HO \ ' 0 HO HPLC-MS (Method (A)): m/z: 305 (M+1); Rt = 4.97 min. Example 552 (General Procedure (E)) 5 3-Hydroxy-7-(4-methoxyphenyl)-naphthalene-2-carboxylic acid O 0, CH3 HO HO HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.68 min. Example 553 (General Procedure (E)) 10 7-(3-Ethoxyphenyl)-3-hydroxynaphthalene-2-carboxylic acid 0 HO 0 HO CH 3 HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 4.89 min. Example 554 (General Procedure (E)) 15 7-Benzo[1,3]dioxol-5-yl-3-hydroxynaphthalene-2-carboxylic acid 0 0I> HO O HO HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 5.61 min. Example 555 (General Procedure (E)) 20 7-Biphenyl-3-yl-3-hydroxynaphthalene-2-carboxylic acid 0 HO
HO
WO 2004/080480 PCT/DK2004/000158 249 HPLC-MS (Method (A)): m/z: 341 (M+1); Rt = 5.45 min. General procedure (F) for preparation of compounds of general formula 15s: O O H 0 H HO H R2 H H 32)_j + J-N(R32)H HO N(R HO HO 5 15 wherein R 3 0 is hydrogen or Cl-C 6 -alkyl and T is as defined above This general procedure (F) is further illustrated in the following example: 10 Example 556 (General procedure (F)) 3-Hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic Acid CH O CH 3 HO N '- N HO 7-Formyl-3-hydroxynaphthalene-2-carboxylic acid (40 mg, 0.19 mmol) (example 535) was 15 suspended in methanol (300 gL). Acetic acid (16 gL, 17 mg, 0.28 mmol) and 4-(2 propyl)aniline (40 gL, 40 mg, 0.30 mmol) were added consecutively, and the resulting mix ture was stirred vigorously at room temperature for 2 hours. Sodium cyanoborohydride (1.0 M in tetrahydrofuran, 300 pL, 0.3 mmol) was added, and the stirring was continued for an other 17 hours. The reaction mixture was poured into 6 N hydrochloric acid (aq.) (6 mL), and 20 the precipitate was filtered off and rinsed with water (3 x 2 mL) to yield the title compound (40 mg) as its hydrochloride salt. No further purification was necessary. 1 H-NMR (DMSO-d 6 ): 610.95 (1H, bs), 8.45 (1H, s), 7.96 (1H, s), 7.78 (1H, d), 7.62 (1H, d), 7.32 (1H, s), 7.13 (2H, bd), 6.98 (2H, bd), 4.48 (2H, s), 2.79 (1H, sept), 1.14 (6H, d); HPLC 25 MS (Method (A)): m/z: 336 (M+1); Rt = 3.92 min.
WO 2004/080480 PCT/DK2004/000158 250 The compounds in the following examples were made using this general procedure (F). Example 557 (General procedure (F)) 7-{[(4-Bromophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid o Br HO N H 5 HO HPLC-MS (Method C): m/z: 372 (M+1); Rt = 4.31min. Example 558 (General procedure (F)) 7-{[(3,5-Dichlorophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid o HO N CI H 10 HO HPLC-MS (Method C): m/z: 362 (M+I1); Rt = 4.75 min. Example 559 (General procedure (F)) 7-{[(Benzothiazol-6-yl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid 0 N HO N N S H 15 HO HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.43 min. Example 560 (General procedure (F)) 3-Hydroxy-7-{[(q uinolin-6-yl)amino]methyl}naphthalene-2-carboxylic Acid NN 0 HO NiN rH 20 HO HPLC-MS (Method C): m/z: 345 (M+1); Rt = 2.26 min. Example 561 (General procedure (F)) 3-Hydroxy-7-{[(4-methoxyphenyl)amino]methyl}naphthalene-2-carboxylic Acid WO 2004/080480 PCT/DK2004/000158 251 0 -Oc~ H O' CH 3 HO ' 'N N " H HO HPLC-MS (Method C): m/z: 324 (M+1); Rt = 2.57min. Example 562 (General procedure (F)) 5 7-{[(2,3-Dihyd robenzofuran-5-ylmethyl)amino]methyl}-3-hyd roxynaphthalene-2-carboxylic Acid 0 HO H HO 0 HPLC-MS (Method C): mlz: 350 (M+1); Rt = 2.22 min. 10 Example 563 (General procedure (F)) 7-{[(4-Chlorobenzyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid 0 HOH N Cl HI otc HO C HPLC-MS (Method C): m/z: 342 (M+1); Rt = 2.45 min. 15 Example 564 (General procedure (F)) 3-Hydroxy-7-{[(naphthalen-1-ylmethyl)amino]methyl}naphthalene-2-carboxylic Acid 0 HOH N HI HO HPLC-MS (Method C): m/z: 357 (M+1); Rt = 2.63 min. 20 Example 565 (General procedure (F)) 7-{[(Biphenyl-2-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid 0 HO N HI 770 HOCI 'N WO 2004/080480 PCT/DK2004/000158 252 HPLC-MS (Method C): m/z: 384 (M+1); Rt = 2.90 min. Example 566 (General procedure (F)) 3-Hydroxy-7-{[(4-phenoxybenzyl)amino]methyl}naphthalene-2-carboxylic Acid 0 HO N HH 5 HO 0 HPLC-MS (Method C): m/z: 400 (M+1); Rt = 3.15 min. Example 567 (General procedure (F)) 3-Hydroxy-7-{[(4-methoxybenzyl)amino]methyl}naphthalene-2-carboxylic Acid 0 HO N HI 10 HO 0 HPLC-MS (Method C): m/z: 338 (M+1); Rt = 2.32 min. General procedure (G) for preparation of compounds of general formula le: O 0 HO N N-J +(C,-C 6 -alkanoyl) 2 0 ----- HO N H HO H HO O (0o-Cs-alkyl) 15 16 wherein J is as defined above and the moiety (CI-C 6 -alkanoyl) 2 0 is an anhydride. The general procedure (G) is illustrated by the following example: 20 Example 568 (General procedure (G)) N-Acetyl-3-hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic Acid CH a CHz HO N 3 HO 0 CH3 WO 2004/080480 PCT/DK2004/000158 253 3-Hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic acid (25 mg, 0.07 mmol) (example 556) was suspended in tetrahydrofuran (200 pL). A solution of sodium hy drogencarbonate (23 mg, 0.27 mmol) in water (200 pL) was added followed by acetic anhy dride (14 1 iL, 15 mg, 0.15 mmol). The reaction mixture was stirred vigorously for 65 hours at 5 room temperature before 6 N hydrochloric acid (4 mL) was added. The precipitate was fil tered off and rinsed with water (3 x 1 mL) to yield the title compound (21 mg). No further puri fication was necessary. 1 H-NMR (DMSO-d 6 ): S10.96 (1H, bs), 8.48 (1H, s), 7.73 (1H, s), 7.72 (1H, d), 7.41 (1H, dd), 10 7.28 (1H, s), 7.23 (2H, d), 7.18 (2H, d), 4.96 (2H, s), 2.85 (1H, sept), 1.86 (3H, s), 1.15 (6H, d); HPLC-MS (Method (A)): m/z: 378 (M+1); Rt = 3.90 min. The compounds in the following examples were prepared in a similar fashion. 15 Example 569 (General procedure (G)) N-Acetyl-7-{[(4-bromophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid OH Br HO -' N HO 0 O~CH3 HPLC-MS (Method C): m/z: 414 (M+1); Rt = 3.76 min. 20 Example 570 (General procedure (G)) N-Acetyl-7-{[(2,3-dihydrobenzofuran-5-ylmethyl)amino]methyl)}-3-hydroxynaphthalene-2 carboxylic Acid 0 HO HO 0, OH 3 HPLC-MS (Method C): m/z: 392 (M+1); Rt = 3.26 min. 25 Example 571 (General procedure (G)) N-Acetyl-7-{[(4-chlorobenzyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid 0 HO N HO 0 OCH 3 1 WO 2004/080480 PCT/DK2004/000158 254 HPLC-MS (Method C): mlz: 384 (M+1); Rt = 3.67 min. Compounds of the invention may also include tetrazoles: Example 572 5 5-(3-(Naphthalen-2-yloxymethyl)-phenyl)-1 H-tetrazole oH 0 N N N To a mixture of 2-naphthol (10 g, 0.07 mol) and potassium carbonate (10 g, 0.073 mol) in acetone (150 mL), alpha-bromo-m-tolunitril (13.6 g, 0.07 mol) was added in portions. The reaction mixture was stirred at reflux temperature for 2.5 hours. The cooled reaction mixture 10 was filtered and evaporated in vacuo affording an oily residue (19 g) which was dissolved in diethyl ether (150 mL) and stirred with a mixture of active carbon and MgSO 4 for 16 hours. The mixture was filtered and evaporated in vacuo affording crude 18.0 g (100 %) of 3 (naphthalen-2-yloxymethyl)-benzonitrile as a solid. 12 g of the above benzonitrile was recrystallised from ethanol (150 mL) affording 8.3 g (69 15 %) of 3-(naphthalen-2-yloxymethyl)-benzonitrile as a solid. M.p. 60 - 61 oC. Calculated for C 1 8
H
13 NO: C, 83.37 %; H, 5.05 %; N, 5.40 %; Found 20 C, 83.51 %; H, 5.03 %; N, 5.38 %. To a mixture of sodium azide (1.46 g, 22.5 mmol) and ammonium chloride (1.28 g, 24.0 mmol) in dry dimethylformamide (20 mL) under an atmosphere of nitrogen, 3-(naphthalen-2 yloxymethyl)-benzonitrile (3.9 g, 15 mmol) was added and the reaction mixture was stirred at 25 125 oC for 4 hours. The cooled reaction mixture was poured on to ice water (300 mL) and acidified to pH = 1 with 1 N hydrochloric acid. The precipitate was filtered off and washed with water, dried at 100 oC for 4 hours affording 4.2 g (93 %) of the title compound. M.p. 200 - 202 oC. 30 Calculated for C 1 8
H
14
N
4 0: C, 71.51 %; H, 4.67 %; N, 18.54 %; Found C, 72.11 %; H, 4.65 %; N, 17.43 %.
WO 2004/080480 PCT/DK2004/000158 255 'H NMR (400 MHz, DMSO-d 6 ) 6sH 5.36 (s, 2H), 7.29 (dd, 1H), 7.36 (dt, 1H), 7.47 (m, 2H), 7.66 (t, 1 H), 7.74 (d, 1H), 7.84 (m, 3H), 8.02 (d, 1H), 8.22 (s, 1H). 5 Example 573 N-(3-(Tetrazol-5-yl)phenyl)-2-naphtoic acid amide 0 N N HN HN 2-Naphtoic acid (10 g, 58 mmol) was dissolved in dichloromethane (100 mL) and NN 10 dimethylformamide (0.2 mL) was added followed by thionyl chloride (5.1 ml, 70 mmol). The mixture was heated at reflux temperature for 2 hours. After cooling to room temperature, the mixture was added dropwise to a mixture of 3-aminobenzonitril (6.90 g, 58 mmol) and triethyl amine (10 mL) in dichloromethane (75 mL). The resulting mixture was stirred at room tem perature for 30 minutes. Water (50 mL) was added and the volatiles was exaporated in. 15 vacuo. The resulting mixture was filtered and the filter cake was washed with water followed by heptane (2 x 25 mL). Drying in vacuo at 50 aC for 16 hours afforded 15.0 g (95 %) of N-(3 cyanophenyl)-2-naphtoic acid amide. M.p. 138-140 C 20 The above naphthoic acid amide (10 g, 37 mmol) was dissolved in N,N-dimethylformamide (200 mL) and sodium azide (2.63 g, 40 mmol) and ammonium chloride (2.16 g, 40 mmol) were added and the mixture heated at 125 'C for 6 hours. Sodium azide (1.2 g) and ammo nium chloride (0.98 g) were added and the mixture heated at 125 oC for 16 hours. After cool 25 ing, the mixture was poured into water (1.5 I) and stirred at room temperature for 30 minutes. The solid formed was filtered off, washed with water and dried in vacuo at 50 oC for 3 days affording 9.69 g (84 %) of the title compound as a solid which could be further purified by treatment with ethanol at reflux temperature. 30 'H NMR (200 MHz, DMSO-de): SH 7.58-7.70 (m, 3H), 7.77 (d, 1H), 8.04-8.13 (m, 5H), 8.65 (d, 1 H), 10.7 (s, 1H).
WO 2004/080480 PCT/DK2004/000158 256 Calculated for Ca 18
H
13
N
5 0O, 0.75 H 2 0: C, 65.74 %; H, 4.44 %; N, 21.30 %. Found: C, 65.58 %; H, 4.50 %; N, 21.05%. 5 Example 574 5-[3-(Biphenyl-4-yloxymethyl)phenyl]-1 H-tetrazole ,N N NH o To a solution of 4-phenylphenol (10.0 g, 59 mmol) in dry N,N-dimethyl-formamide (45 mL) kept under an atmosphere of nitrogen, sodium hydride (2.82 g, 71 mmol, 60 % dispersion in 10 oil) was added in portions and the reaction mixture was stirred until gas evolution ceased. A solution of m-cyanobenzyl bromide (13 g, 65 mmol) in dry N,N-dimethylformamide (45 mL) was added dropwise and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was poured on to ice water (150 mL). The precipitate was filtered of and washed with 50 % ethanol 15 (3 x 50 mL), ethanol (2 x 50 mL), diethyl ether (80 mL), and dried in vacuo at 50 OC for 18 hours affording crude 17.39 g of 3-(biphenyl-4-yloxymethyl)-benzonitrile as a solid. 1 H NMR (200 MHz, CDCl 3 ) 6 SH 5.14 (s, 2H), 7.05 (m, 2H), 7.30 - 7.78 (m, 11H). 20 To a mixture of sodium azide (2.96 g, 45.6 mmol) and ammonium chloride (2.44 g, 45.6 mmol) in dry N,N-dimethylformamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl 4-yloxymethyl)-benzonitrile (10.0 g, 35.0 mmol) was added and the reaction mixture was stirred at 125 °C for 18 hours. The cooled reaction mixture was poured on to a mixture of 1N 25 hydrochloric acid (60 mL) and ice water (500 mL). The precipitate was filtered off and washed with water (3 x 100 mL), 50 % ethanol (3 x 100 mL), ethanol (50 mL), diethyl ether (50 mL), ethanol (80 mL), and dried in vacuo at 50 oC for 18 hours affording 8.02 g (70 %) of the title compound. 30 1 H NMR (200 MHz, DMSO-d 6 ) 5 H 5.31 (s, 2H), 7.19 (m, 2H), 7.34 (m, 1H), 7.47 (m, 2H), 7.69 (m, 6H), 8.05 (dt, 1H), 8.24 (s, 1H).
WO 2004/080480 PCT/DK2004/000158 257 Example 575 5-(3-Phenoxymethyl)-phenyl)-tetrazole 0 ~N. N- N H 5 3-Bromomethylbenzonitrile (5.00 g, 25.5 mmol) was dissolved in N,N-dimethylformamide (50 mL), phenol (2.40 g, 25.5 mmol) and potassium carbonate (10.6 g, 77 mmol) were added. The mixture was stirred at room temperature for 16 hours. The mixture was poured into wa ter (400 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic extracts were washed with water (2 x 100 mL), dried (MgSO 4 ) and evaporated in vacuo to afford 5.19 10 g (97 %) 3-(phenoxymethyl)benzonitrile as an oil. TLC: Rf = 0.38 (Ethyl acetate/heptane = 1:4) The above benzonitrile (5.19 g, 24.8 mmol) was dissolved in N,N-dimethylformamide (100 15 mL) and sodium azide (1.93 g, 30 mmol) and ammonium chloride (1.59 g, 30 mmol) were added and the mixture was heated at 140 oC for 16 hours. After cooling, the mixture was poured into water (800 mL). The aqeous mixture was washed with ethyl acetate (200 mL). The pH of the aqueous phase was adjusted to 1 with 5 N hydrochloric acid and stirred at room temperature for 30 minutes. Filtration, washing with water and drying in vacuo at 50 aC 20 afforded 2.06 g (33 %) of the title compound as a solid. 1 H NMR (200 MHz, CDCl 3 + DMSO-d) 6 H 5.05 (s, 2H), 6.88 (m, 3H), 7.21 (m, 2H), 7.51 (m, 2H), 7.96 (dt, 1H), 8.14 (s, 1H). Example 576 25 5-[3-(Biphenyl-4-ylmethoxy)phenyl]-1 H-tetrazole H 'NN To a solution of 3-cyanophenol (5.0 g, 40.72 mmol) in dry N,N-dimethylformamide (100 mL) kept under an atmosphere of nitrogen, sodium hydride (2 g, 48.86 mmol, 60 % dispersion in oil) was added in portions and the reaction mixture was stirred until gas evolution ceased. p- WO 2004/080480 PCT/DK2004/000158 258 Phenylbenzyl chloride (9.26 g, 44.79 mmol) and potassium iodide (0.2 g, 1.21 mmol) were added and the reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was poured on to a mixture of saturated sodium carbonate (100 mL) and ice water (300 mL). The precipitate was filtered of and washed with water (3 x 100 mL), n-hexane (2 x 5 80 mL) and dried in vacuo at 50 °C for 18 hours affording 11.34 g (98 %) of 3-(biphenyl-4 ylmethoxy)-benzonitrile as a solid. To a mixture of sodium azide (2.37 g, 36.45 mmol) and ammonium chloride (1.95 g, 36.45 mmol) in dry N,N-dimethylformamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl 10 4-ylmethoxy)-benzonitrile (8.0 g, 28.04 mmol) was added and the reaction mixture was stirred at 125 OC for 18 hours. To the cooled reaction mixture water (100 mL) was added and the reac tion mixture stirred for 0.75 hour. The precipitate was filtered off and washed with water, 96 % ethanol (2 x 50 mL), and dried in vacuo at 500C for 18 hours affording 5.13 g (56 %) of the 15 title compound. 1 H NMR (200 MHz, DMSO-d 6 ) 8H 5.29 (s, 2H), 7.31 (dd, 1H), 7.37 - 7.77 (m, 12H). Example 577 20 5-[4-(Biphenyl-4-ylmethoxy)-3-methoxyphenyl]-1 H-tetrazol / N-N o H
CH
3 This compound was made similarly as described in example 576. Example 578 25 0 II H \N 0 N-
N
' Example 579 5-(2-Naphtylmethyl)-1 H-tetrazole WO 2004/080480 PCT/DK2004/000158 259 H This compound was prepared similarly as described in example 572, step 2. Example 580 5 5-(1-Naphtylmethyl)-1lH-tetrazole N-N I N N H This compound was prepared similarly as described in example 572, step 2. Example 581 10 5-[4-(Biphenyl-4-yloxymethyl)phenyl]-1 H-tetrazole Oao - N-N H A solution of alpha-bromo-p-tolunitrile (5.00 g, 25.5 mmol), 4-phenylphenol (4.56 g, 26.8 mmol), and potassium carbonate (10.6 g, 76.5 mmol) in N,N-dimethylformamide (75 mL) was stirred vigorously for 16 hours at room temperature. Water (75 mL) was added and the mix 15 ture was stirred at room temperature for 1 hour. The precipitate was filtered off and washed with thoroughly with water. Drying in vacuo over night at 50 oC afforded 7.09 g (97 %) of 4 (biphenyl-4-yloxymethyl)benzonitrile as a solid. The above benzonitrile (3.00 g, 10.5 mmol) was dissolved in N,N-dimethylformamide (50 20 mL), and sodium azide (1.03 g, 15.8 mmol) and ammonium chloride (0.84 g, 15.8 mmol) were added and the mixture was stirred 16 hours at 125 oC. The mixture was cooled to room temperature and water (50 mL) was added. The suspension was stirred overnight, filtered, washed with water and dried in vacuo at 50 °C for 3 days to give crude 3.07 g (89 %) of the title compound. From the mother liquor crystals were colected and washed with water, dried 25 by suction to give 0.18 g (5 %) of the title compound as a solid.
WO 2004/080480 PCT/DK2004/000158 260 'H NMR (200 MHz, DMSO-d 6 ): 6H 5.21 (s, 2H), 7.12 (d, 2H), 7.30 (t, 1H), 7.42 (t, 2H), 7.56 7.63 (m, 6H), 8.03 (d, 2H). Calculated for C 2 0
H
1 6N 4 0, 2H 2 0: 5 C, 65.92 %; H, 5.53 %; N, 15.37 %. Found: C, 65.65 %; H, 5.01 %; N, 14.92 %. Example 582 100 NN 10 H This compound was prepared similarly as described in example 576. Example 583 IHN O. N" 15 Example 584 N=N ~~.H N. NH C-N HI 0 N ON 20 Example 585 WO 2004/080480 PCT/DK2004/000158 261 H NN N= N Example 586 5-(3-(Biphenyl-4-yloxymethyl)-benzyl)-1 H-tetrazole o N N 5 H Example 587 5-(1-Naphthyl)-1H-tetrazole N=N I % 10 This compound was prepared similarly as described in example 572, step 2. Example 588 5-[3-Methoxy-4-(4-methylsulfonylbenzyloxy)phenyl]-1 H-tetrazole " O' / CO O 15 This compound was made similarly as described in example 576. Example 589 5-(2-Naphthyl)-1H-tetrazole J', N N 0 0 I/ 'CH 3 0 15 This compound was made similarly as described in example 576. Example 589 5-(2-Naphthyl)-1 H-tetrazole WO 2004/080480 PCT/DK2004/000158 262 N-N, I N N C:: H This compound was prepared similarly as described in example 572, step 2. Example 590 5 2-Amino-N-(1H-tetrazol-5-yl)-benzamide 0 N-N I,N N NN H H
NH
2 Example 591 5-(4-Hydroxy-3-methoxyphenyl)-1lH-tetrazole N=N I % NN NH
H
3 C 0 10 OH This compound was prepared similarly as described in example 572, step 2. Example 592 4-(2H-Tetrazol-5-ylmethoxy)benzoic acid 0 ~- OH HN ( O& O H 15 'N7N To a mixture of methyl 4-hydroxybenzoate (30.0 g, 0.20 mol), sodium iodide (30.0 g, 0.20 mol) and potassium carbonate (27.6 g, 0.20 mol) in acetone (2000 mL) was added chloroacetonitrile (14.9 g , 0.20 mol). The mixture was stirred at RT for 3 days. Water was added and the mixture was acidified with 1N hydrochloric acid and the mixture was extracted 20 with diethyl ether. The combined organic layers were dried over Na 2
SO
4 and concentrated in vacuo. The residue was dissolved in acetone and chloroacetonitrile (6.04 g,0.08 mol), so- WO 2004/080480 PCT/DK2004/000158 263 dium iodide (12.0 g, 0.08 mol) and potassium carbonate (11.1 g, 0.08 mol) were added and the mixture was stirred for 16 hours at RT and at 60 °C. More chloroacetonitrile was added until the conversion was 97%. Water was added and the mixture was acidified with 1N hy drochloric acid and the mixture was extracted with diethyl ether. The combined organic lay 5 ers were dried over Na 2
SO
4 and concentrated in vacuo to afford methyl 4 cyanomethyloxybenzoate in quantitative yield. This compound was used without further puri fication in the following step. A mixture of methyl 4-cyanomethyloxybenzoate (53.5 g,0.20 mol), sodium azide (16.9 g, 0.26 10 mol) and ammonium chloride (13.9 g, 0.26 mol) in DMF 1000 (mL) was refluxed overnight under N 2 . After cooling, the mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over Na 2
SO
4 and concentrated in vacuo, to afford methyl 4-(2H-tetrazol-5 ylmethoxy)benzoate. This compound was used as such in the following step. 15 Methyl 4-(2H-Tetrazol-5-ylmethoxy)-benzoate was refluxed in 3N sodium hydroxide. The re action was followed by TLC (DCM:MeOH = 9:1). The reaction mixture was cooled, acidified and the product filtered off. The impure product was washed with DCM, dissolved in MeOH, filtered and purified by column chromatography on silica gel (DCM:MeOH = 9:1).The result 20 ing product was recrystallised from DCM:MeOH=95:5. This was repeated until the product was pure. This afforded 13.82 g (30 %) of the title compound. 1 H-NMR (DMSO-d): 4.70 (2H, s), 7.48 (2H, d), 7.73 (2H, d), 13 (1H, bs). 25 Example 593 4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoic acid 0 O \- OH HN S 'NzN To a solution of sodium hydroxide (10.4 g, 0.26 mol) in degassed water (600 mL) was added 4-mercaptobenzoic acid (20.0 g, 0.13 mol). This solution was stirred for 30 minutes. To a 30 solution of potassium carbonate (9.0 g, 65 mmol) in degassed water (400 mL) was added chloroacetonitrile (9.8 g, (0.13 mol) portion-wise. These two solutions were mixed and stirred for 48 hours at RT under N 2 . The mixture was filtered and washed with heptane. The aque- WO 2004/080480 PCT/DK2004/000158 264 ous phase was acidified with 3N hydrochloric acid and the product was filtered off, washed with water and dried, affording 4-cyanomethylsulfanylbenzoic acid (27.2 g, 88%). This com pound was used without further purification in the following step. 5 A mixture of 4-cyanomethylsulfanylbenzoic acid (27.2 g, 0.14 mol), sodium azide (11.8 g, 0,18 mol) and ammonium chloride (9.7 g, 0.18 mol) in DMF (1000 mL) was refluxed over night under N 2 . The mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with diethyl ether. The combined organic phases were washed with brine, dried over Na 2
SO
4 and concentrated in vacuo. Water was added and the precipitate 10 was filtered off. The aqueous layer was concentrated in vacuo, water was added and the precipitate filtered off. The combined impure products were purified by column chromatogra phy using DCM:MeOH = 9:1 as eluent, affording the title compound (5.2 g, 16%). 1 H-NMR (DMSO-d 6 ): 5.58 (2H, s), 7.15 (2H, d), 7.93 (2H, d), 12.7 (1H, bs). 15 Example 594 3-(2H-Tetrazol-5-yl)-9H-carbazole HN 'N HN N H 3-Bromo-9H-carbazole was prepared as described by Smith et al. in Tetrahedron 1992, 48, 20 7479-7488. A solution of 3-bromo-9H-carbazole (23.08 g, 0.094 mol) and cuprous cyanide (9.33 g, 0.103 mol) in N-methyl-pyrrolidone (300 ml) was heated at 200 OC for 5 h. The cooled reaction mix ture was poured on to water (600 ml) and the precipitate was filtered off and washed with ethyl acetate (3 x 50 ml). The filtrate was extracted with ethyl acetate (3 x 250 ml) and the 25 combined ethyl acetate extracts were washed with water (150 ml), brine (150 ml), dried (MgSO 4 ) and concentrated in vacuo. The residue was crystallised from heptanes and recrys tallised from acetonitrile (70 ml) affording 7.16 g (40 %) of 3-cyano-9H-carbazole as a solid. M.p. 180- 181 OC. 30 3-Cyano-9H-carbazole (5.77 g, 30 mmol) was dissolved in N,N-dimethylformamide (150 ml), and sodium azide (9.85 g, 152 mmol), ammonium chloride (8.04 g, 150 mmol) and lithium WO 2004/080480 PCT/DK2004/000158 265 chloride (1.93 g, 46 mmol) were added and the mixture was stirred for 20 h at 125 oC. To the reaction mixture was added an additional portion of sodium azide (9.85 g, 152 mmol) and ammonium chloride (8.04 g, 150 mmol) and the reaction mixture was stirred for an additional 24 h at 125 oC. The cooled reaction mixture was poured on to water (500 ml). The suspen 5 sion was stirred for 0.5 h, and the precipitate was filtered off and washed with water (3 x 200 ml) and dried in vacuo at 50 oC. The dried crude product was suspended in diethyl ether (500 ml) and stirred for 2 h, filtered off and washed with diethyl ether (2 x 200 ml) and dried in vacuo at 50 oC affording 5.79 g (82 %) of the title compound as a solid. 10 1 H-NMR (DMSO-d 6 ): 511.78 (1H, bs), 8.93 (1H, d), 8.23 (1H, d), 8.14 (1H, dd), 7.72 (1H, d), 7.60 (1H, d), 7.49 (1H, t), 7.28 (1H, t); HPLC-MS (Method C): m/z: 236 (M+1I); Rt = 2.77 min. The following commercially available tetrazoles do all bind to the His B10 Zn 2 + site of the insulin hexamer: 15 Example 595 5-(3-Tolyl)-1H-tetrazole
CH
3 I. N N-N H 20 Example 596 5-(2-Bromophenyl)tetrazole H Br N-N Example 597 25 5-(4-Ethoxalylamino-3-nitrophenyl)tetrazole WO 2004/080480 PCT/DK2004/000158 266 H 0 0 HN 0 I N N N HN- // Example 598 5 cNH Example 599 N, NH 0 '0' o 10 Example 600 N-N /N NH Cl 15 Example 601 15 Example 601 WO 2004/080480 PCT/DK2004/000158 267 F F 7H F F Example 602 Tetrazole H N r N 5 N Example 603 5-Methyltetrazole H H3N / I N-N 10 Example 604 5-Benzyl-2H-tetrazole HN~ 15 Example 605 4-(2H-Tetrazol-5-yl)benzoic acid 0 HOH Example 606 20 5-Phenyl-2H-tetrazole WO 2004/080480 PCT/DK2004/000158 268 Example 607 5-(4-Chlorophenylsulfanylmethyl)-2H-tetrazole HN--I S C 5 Example 608 5-(3-Benzyloxyphenyl)-2H-tetrazole H" O 10 Example 609 2-Phenyl-6-(1 H-tetrazol-5-yl)-chromen-4-one N-H N 0 N NO Example 610 15 CI H2C HN N Example 611 WO 2004/080480 PCT/DK2004/000158 269 F -N F N Example 612 o 0 H.C - -_ 0 H o / N N 5 H3C Example 613 N NH N-N CH
H
2 F /N H 15 Example 615 5-(4-Bromo-phenyl)-1 H-tetrazole WO 2004/080480 PCT/DK2004/000158 270 Br N- H Example 616 cI HNA, 5 N Example 617 H H/ C 10 Example 618 H
N
4N 15 Example 619 7 N Example 620 20 WO 2004/080480 PCT/DK2004/000158 271 0 o= / H NN HO O N 0> HzC Example 621 rO N-_ O 0
-
NN 5 H Example 622 N N HO 0 OH 10 Example 623 F 15 Example 624 WO 2004/080480 PCT/DK2004/000158 272 W- H 14 0
H
3 C Example 625 Cl- 0 H Ci N 5 H N Example 626 0 0 OH
N
HC 10 Example 627 C)HNNN
H
3 C N N N/ / H 15 Example 628 0 0 0 H t / NN Example 629 20 WO 2004/080480 PCT/DK2004/000158 273 Br
N-
H HO Br Example 630 5 o Example 631 0 / OCN H \>-N HO N
CH
3 10 Example 632 NN H Cl-f 3 SCH3 15 Example 633 WO 2004/080480 PCT/DK2004/000158 274 CI o
N
N 'IN Example 634 H N N F ~N 5 Example 635
H
3 C 0 O NH H \N NN 10 Example 636 NN 0 ENplN o 15 Example 637 WO 2004/080480 PCT/DK2004/000158 275 H N / H N O \ N-N O O Example 638 o 0 NN N- // 5 Example 639 N N FHN/ 0 10 Example 640 NN HN'N 0 N H N 15 Example 641 WO 2004/080480 PCT/DK2004/000158 276 0 H+-- /Cl N H 0 Example 642 HNy N NH 5 c o Example 643 H H 0 0 Br 10 Example 644 CI C 0 H HN N Y N
NN
WO 2004/080480 PCT/DK2004/000158 277 Example 645
H
3 C N-N\ 0~ N H 5 Example 646 5-(2,6-Dichlorobenzyl)-2H-tetrazole Cl General procedure (H) for preparation of compounds of general formula 17: NaCBH 3 .. 0 ~ HOAc HN K H DMF H NKH N'N M-NH 2 H T- N N M-N T H 10 17 wherein K, M, and T are as defined above. The reaction is generally known as a reductive alkylation reaction and is generally performed by stirring an aldehyde with an amine at low pH (by addition of an acid, such as acetic acid or 15 formic acid) in a solvent such as THF, DMF, NMP, methanol, ethanol, DMSO, dichloro methane, 1,2-dichloroethane, trimethyl orthoformate, triethyl orthoformate, or a mixture of two or more of these. As reducing agent sodium cyano borohydride or sodium triacetoxy borohydride may be used. The reaction is performed between 20 0 C and 120 0 C, preferably at room temperature. 20 When the reductive alkylation is complete, the product is isolated by extraction, filtration, chro matography or other methods known to those skilled in the art. The general procedure (H) is further illustrated in the following example 647: 25 WO 2004/080480 PCT/DK2004/000158 278 Example 647 (General procedure (H)) Biphenyl-4-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine NzN H HR~ N N A solution of 5-(3-aminophenyl)-2H-tetrazole (example 874, 48 mg, 0.3 mmol) in DMF (250 5 jiL) was mixed with a solution of 4-biphenylylcarbaldehyde (54 mg, 0.3 mmol) in DMF (250 PL) and acetic acid glacial (250 g.L) was added to the mixture followed by a solution of so dium cyano borohydride (15 mg, 0.24 mmol) in methanol (250 p.L). The resulting mixture was shaken at room temperature for 2 hours. Water (2 mL) was added to the mixture and the re sulting mixture was shaken at room temperature for 16 hours. The mixture was centrifugated 10 (6000 rpm, 10 minutes) and the supernatant was removed by a pipette. The residue was washed with water (3 mL), centrifugated (6000 rpm, 10 minutes) and the supernatant was removed by a pipette. The residue was dried in vacuo at 40 oC for 16 hours to afford the title compound as a solid. 15 HPLC-MS (Method C): m/z: 328 (M+1), 350 (M+23); Rt 4.09 min. Example 648 (General procedure (H)) Benzyl-[3-(2H-tetrazol-5-yl)phenyl]amine NzN H 5: HNN/. 20 HPLC-MS (Method D): mlz: 252 (M+1); Rt = 3,74 min. Example 649 (General procedure (H)) (4-Methoxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine HNN H " O'CH 3 25 HPLC-MS (Method D): m/z: 282,2 (M+); Rt = 3,57min. 25 HPLC-MS (Method D): mfz: 282,2 (M+1); Rt = 3,57min.
WO 2004/080480 PCT/DK2004/000158 279 Example 650 (General procedure (H)) 4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenol N:N N ,- OH HN. H HPLC-MS (Method D): m/z: 268,4 (M+1); Rt = 2,64 min. 5 Example 651 (General procedure (H)) (4-Nitrobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine O II+ HNN H - N N 'N , HPLC-MS (Method D): m/z: 297,4 (M+1); Rt = 3,94 min. 10 Example 652 (General procedure (H)) (4-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine NzN H Cl HN. N Cl N ' HPLC-MS (Method D): m/z: 287,2 (M+1); Rt = 4,30 min. 15 Example 653 (General procedure (H)) (2-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine HNN N N 'N _p CI HPLC-MS (Method D): m/z: 286 (M+1); Rt = 4,40 min. 20 Example 654 (General procedure (H)) (4-Bromobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine WO 2004/080480 PCT/DK2004/000158 280 SN N Br HPLC-MS (Method D): m/z:332 (M+1); Rt = 4,50 min. Example 655 (General procedure (H)) 5 (3-Benzyloxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine HNO HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,94 min. Example 656 (General procedure (H)) 10 Naphthalen-1 -ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine FIN'~N H I N~j N 1-6 NN HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min. Example 657 (General procedure (H)) 15 Naphthalen-2-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine 'N=N H HNN H . ~ N N HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,60 min. Example 658 (General procedure (H)) 20 4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid 0 ~ OH H1 N N OH HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,24 min.
WO 2004/080480 PCT/DK2004/000158 281 Example 659 (General procedure (H)) [3-(2H-Tetrazol-5-yl)-phenyl]-[3-(3-trifluoromethyl-phenoxy)benzyl]amine F I 5 HPLC-MS (Method D): m/z: 412 (M+1); Rt = 5,54 min. Example 660 (General procedure (H)) (3-Phenoxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine HN H o, HN ~ N
O
N iia01 10 HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,04 min. Example 661 (General procedure (H)) (4-Phenoxy-benzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine WiN: N H I 1z: HNN N " 15 HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,00 min. Example 662 (General procedure (H)) (4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenoxy)acetic acid 0 H , N H O--I O H 20 HPLC-MS (Method D): m/z: 326 (M+1); Rt = 3,10 min. Example 663 (General procedure (H)) (4-Benzyloxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine WO 2004/080480 PCT/DK2004/000158 282 N HPLC-MS (Method D): m/z: 358 (M+1I); Rt = 4,97 min. Example 664 (General procedure (H)) 5 3-(4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenyl)acrylic acid 0 HNN H OH .N - N N. HPLC-MS (Method D): m/z: 322 (M+1); Rt = 3,60 min. Example 665 (General procedure (H)) 10 Dimethyl-(4-{[3-(2H-tetrazol-5-yl)phenylamino]methyl}naphthalen-1 -yl)amine I CH3 ,NN H N'CH 3 HN. N \. N N. HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,07 min. Example 666 (General procedure (H)) 15 (4'-Methoxybiphenyl-4-ylmethyl)-[3-(2H-tetrazol-5-yl)phenyl]amine CH3 0 H N HN H HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,97 min. Example 667 (General procedure (H)) 20 (2'-Chlorobiphenyl-4-ylmethyl)-[3-(2H-tetrazol-5-yl)phenyl]amine WO 2004/080480 PCT/DK2004/000158 283 HN.N N Cl N N HPLC-MS (Method D): m/z: 362 (M+1); Rt = 5,27 min. Example 668 (General procedure (H)) 5 Benzyl-[4-(2H-tetrazol-5-yl)phenyl]amine N N HNN N For preparation of starting material, see example 875. HPLC-MS (Method D): m/z: 252 (M+1); Rt = 3,97 min. 10 Example 669 (General procedure (H)) (4-Methoxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine N=N HN.N N O'C. H
CH
3 HPLC-MS (Method D): m/z: 282 (M+1); Rt = 3,94 min. 15 Example 670 (General procedure (H)) 4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenol ,NzN HN.l N OH N N HPLC-MS (Method D): m/z: 268 (M+1); Rt = 3,14 min. 20 Example 671 (General procedure (H)) (4-Nitrobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine WO 2004/080480 PCT/DK2004/000158 284 ,N1 N HN N+ N . - H N , 0 N HPLC-MS (Method D): m/z: (M+1); Rt = 3,94 min. Example 672 (General procedure (H)) 5 ( 4 -Chlorobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
,N--
N HN. H NN N H C I N Cl HPLC-MS (Method D): m/z: (M+1); Rt = 4,47 min. Example 673 (General procedure (H)) 10 ( 2 -Chlorobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine HN
=
N N N . CI HPLC-MS (Method D): m/z: 286 (M+1); Rt = 4,37 min. Example 674 (General procedure (H)) 15 (4-Bromobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine N Br
,N~
N HPLC-MS (Method D): m/z: 331 (M+1); Rt = 4,57 min. Example 675 (General procedure (H)) 20 (3-Benzyloxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine HN
-
WO 2004/080480 PCT/DK2004/000158 285 HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,07min. Example 676 (General procedure (H)) Naphthalen-1 -ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine ,NzN HN. N H N 5 HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min. Example 677 (General procedure (H)) Naphthalen-2-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine
,NN
N 10 HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min. Example 678 (General procedure (H)) Biphenyl-4-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine N N N N\N. 15 HPLC-MS (Method D): m/z: 328 (M+1); Rt = 5,07 min. Example 679 (General procedure (H)) 4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid H N N .O 20 OH 20 D HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,34 min.
WO 2004/080480 PCT/DK2004/000158 286 Example 680 (General procedure (H)) [4-(2H-Tetrazol-5-yl)phenyl]-[3-(3-trifluoromethylphenoxy)benzyl]amine ,NzN HNN F HPLC-MS (Method D): m/z: 412 (M+1); Rt = 5,54 min. 5 Example 681 (General procedure (H)) (3-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine ,N N HNN NN O HPLC-MS (Method D): m/z: 344 (M+1); Rt= 5,07 min. 10 Example 682 (General procedure (H)) (4-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)-phenyl]-amine ,N N H N .N N O HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,03 min. 15 Example 683 (General procedure (H)) 3-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid N N HN~N OH HPLC-MS (Method D): m/z: 286 (M+1); Rt = 3,47 min. 20 Example 684 (General procedure (H)) (4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenoxy)acetic acid WO 2004/080480 PCT/DK2004/000158 287 N::N HN. 0
N
N HPLC-MS (Method D): m/z: 326 (M+1); Rt = 3,40 min. Example 685 (General procedure (H)) 5 ( 4 -Benzyloxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine N NH HNN HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,14 min. Example 686 (General procedure (H)) 10 3 -(4-{[4-(2H-Tetrazol-5-yl)phenylamino]methyl}phenyl)acrylic acid ,N N HN.N N OH N N . HPLC-MS (Method D): m/z: 322 (M+1); Rt = 3,66 min. Example 687 (General procedure (H)) 15 Dimethyl-(4-{[4-(2H-tetrazol-5-yl)phenylamino]methyl}naphthalen-1 -yl)amine ,N N. HN'NzN CH3 H N CH 3 N \ HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,10 min. Example 688 (General procedure (H)) 20 (4'-Methoxybiphenyl-4-ylmethyl)-[4-(2H-tetrazol-5-yl)phenyl]amine CH3 ,NzN 0 HN, HN N O HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,04 min.
WO 2004/080480 PCT/DK2004/000158 288 Example 689 (General procedure (H))
(
2 '-Chlorobiphenyl-4-ylmethyl)-[4-(2H-tetrazol-5-yl)-phenyl]-amine NN HN H N \, oz CI 5 HPLC-MS (Method D): m/z: 362 (M+1); Rt = 5,30 min. General procedure (1) for preparation of compounds of general formula Is: HOAt HNN EDAC HN ON H2T DMF H N N H IN--H 0 1>K kH N I. + H2 NT HN:r T_, OH 0 18 wherein K, M and T are as defined above. 10 This procedure is very similar to general procedure (A), the only difference being the carbox ylic acid is containing a tetrazole moiety. When the acylation is complete, the product is iso lated by extraction, filtration, chromatography or other methods known to those skilled in the art. 15 The general procedure (I) is further illustrated in the following example 690: Example 690 (General procedure (I)) 4-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid N--N HN' N HO ~- N 0 0r OH 0 20 To a solution of 4-(2H-tetrazol-5-yl)benzoic acid (example 605, 4 mmol) and HOAt (4.2 mmol) in DMF (6 mL) was added 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochlo ride (4.2 mmol) and the resulting mixture was stirred at room temperature for 1 hour. An alquot of this HOAt-ester solution (0.45 mL) was mixed with 0.25 mL of a solution of 4 aminobenzoic acid (1.2 mmol in 1 mL DMF). (Anilines as hydrochlorides can also be utilised, WO 2004/080480 PCT/DK2004/000158 289 a slight excess of triethylamine was added to the hydrochloride suspension in DMF prior to mixing with the HOAt-ester.) The resulting mixture was shaken for 3 days at room tempera ture. 1N hydrochloric acid (2 mL) was added and the mixture was shaken for 16 hours at room temperature. The solid was isolated by centrifugation (alternatively by filtration or ex 5 traction) and was washed with water (3 mL). Drying in vacuo at 40 °C for 2 days afforded the title compound. HPLC-MS (Method D): m/z: 310 (M+1); Rt = 2.83 min. 10 Example 691 (General procedure (I)) 3-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid 15 Example 692 (General procedure (1)) 3-{4-[4-(2H-Tetrazol-5-yl)benzoylamino]phenyl}acrylic acid NN HNNH N H 0 ON OH 0O HPLC-MS (Method D): m/z: 31036 (M+1); Rt = 3.1089 min. 1520 Example 693 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-yl)benzoylamino]phenyl)ac}propionylic acid NzN HN H O -OH O 0 HPLC-MS (Method D): m/z: 338 (M+1); Rt = 2.9710 min. 25 Example 694 (General procedure (I)) 3-Methoxy-4-[4-(2H-tetrazol-5-yl)benzoylamino]bphenyl}propionzoic acid HR. N H o 10__ OH 0 HPLC-MS (Method 0): mlz: 338 (M+1); Rt = 2.97 min. 25 Example 694 (General procedure (1)) 3-Methoxy-4-[4-(2H-tetrazol-5-y)benzoylamino]benzoic acid WO 2004/080480 PCT/DK2004/000158 290 N= N H NN C O - OH O 0 HPLC-MS (Method D): m/z: 340 (M+1); Rt = 3.03 min. Example 695 (General procedure (I)) 5 N-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-yl)benzamide N o o HPLC-MS (Method D): m/z: 372 (M+1); Rt = 4.47 min. Example 696 (General procedure (I)) 10 N-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-yl)benzamide NzN HK N-- O HNN H HPLC-MS (Method 0): m/z: 358 (M+1); Rt = 4.50 min. Example 697 (General procedure (I)) 15 N-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-yl)benzamide HN NHK N NH 0 Nj-C HPLC-MS (Method D): m/z: 354 (M+1); Rt = 4.60 min. Example 698 (General procedure (I)) 20 N-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-yl)benzamide WO 2004/080480 PCT/DK2004/000158 291 N. NHK N H rCN 0 HPLC-MS (Method D): m/z: 383 (M+1); Rt = 4.60 min. Example 699 (General procedure (I)) 5 N-Phenyl-4-(2H-tetrazol-5-yl)benzamide .NN H4H HNNL[ H HPLC-MS (Method D): m/z: 266 (M+1); Rt = 3.23 min. Example 700 (General procedure (I)) 10 4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]benzoic acid NzN HNN O' H N O S OH 0 The starting material was prepared as described in example 592. HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.83 min. 15 Example 701 (General procedure (I)) 3-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]benzoic acid NzN HR HN N OH 0 HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.90 min. 20 Example 702 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]phenyl}acrylic acid WO 2004/080480 PCT/DK2004/000158 292 NzN H N , O 0 N OH 0 HPLC-MS (Method D): m/z: 366 (M+1); Rt = 3.07 min. Example 703 (General procedure (I)) 5 3-{4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]phenyl}propionic acid -Y N O N ,OH 0 HPLC-MS (Method D): m/z: 368 (M+1); Rt = 2.97 min. Example 704 (General procedure (I)) 10 3-Methoxy-4-[4-(2H-tetrazol-5-ylmethoxy)benzoylamino]benzoic acid ,NzN H cNHN 0 K OH O 0 HPLC-MS (Method D): m/z: 370 (M+1); Rt = 3.07 min. Example 705 (General procedure (I)) 15 N-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-ylmethoxy)benzamide ,N=N HNoo HK 0 HPLC-MS (Method D): m/z: 402 (M+1); Rt = 4.43 min. Example 706 (General procedure (I)) 20 N-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-ylmethoxy)benzamide NzN HN O H HRN N H -Q N_ ,0 WO 2004/080480 PCT/DK2004/000158 293 HPLC-MS (Method D): m/z: 388 (M+1); Rt = 4.50 min. Example 707 (General procedure (I)) N-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethoxy)benzamide NzN HN O ~-( N j :) __ 5 HPLC-MS (Method D): m/z: 384 (M+1); Rt = 4.57 min. Example 708 (General procedure (I)) N-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-ylmethoxy)benzamide HPLC-MS (Method D): m/z: 413 (M+1); Rt = 4.57 min. Example 709 (General procedure (I)) N-Phenyl-4-(2H-tetrazol-5-ylmethoxy)benzamide HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3.23 main. Example 710 (General procedure (I)) 4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid NzN H OH o-'I H 1kH 100 The starting material was prepared as described in example 593. HPLC-MS (Method D): m/z: 4356 (M+1); Rt = 2.9357 min. Example 709 (General procedure (1)) N-Phenyl-4-(2H-tetrazol-5-ylmethoxy)belzamfide N=N NN HPLC-MS(Method D): mlz: 296 (M+1); Rt = 3.23 min. Example 710 (General procedure (1)) 4-[4-(2H-Tetrazol-5-ymethylsulfanyl)bezoymio]belzoic acid H R N N" 0 K-OH 20 0 The starting material was prepared as described in example 593. HPLC-MS (Method D): mlz: 356 (M+1); Rt = 2.93 min.
WO 2004/080480 PCT/DK2004/000158 294 Example 711 (General procedure (I)) 3-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid NN HCN N OH 0 HPLC-MS (Method D): m/z: 356 (M+1); Rt = 3.00 min. 5 Example 712 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]phenyl}acrylic acid SN - OH 0 HPLC-MS (Method D): m/z: 382 (M+1); Rt = 3.26 min. 10 Example 713 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]phenyl}propionic acid W'N HKNJIS H -y NO_ 0 OH O 0 HPLC-MS (Method D): m/z: 384 (M+1); Rt = 3.10 min. 15 Example 714 (General procedure (I)) 3-Methoxy-4-[4-(2H-tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid N=N HN- 'S NH~ O H .OH N SOH 0 HPLC-MS (Method D): m/z: 386 (M+1); Rt = 3.20 min. 20 Example 715 (General procedure (I)) N-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide WO 2004/080480 PCT/DK2004/000158 295 N=N HI&~o o o HPLC-MS (Method D): m/z: 418 (M+1); Rt = 4.57 min. Example 716 (General procedure (I)) 5 N-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide NzN H NN S HPLC-MS (Method 0): m/z: 404 (M+1); Rt = 4.60 min. Example 717 (General procedure (I)) 10 N-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide N-N H NN-'S" H HPLC-MS (Method D): m/z: 400 (M+1); Rt = 4.67 min. Example 718 (General procedure (I)) 15 N-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide NzN HKN. N 'S H rCH 3 -Y N-CN s HPLC-MS (Method D): m/z: 429 (M+1); Rt = 4.67 min. Example 719 (General procedure (I)) 20 N-Phenyl-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide -NzN HN. N N,0 -0 WO 2004/080480 PCT/DK2004/000158 296 HPLC-MS (Method D): m/z: 312 (M+1); Rt = 3.40 min. General procedure (J) for solution phase preparation of amides of general formula 19: NzN N N N. INN H H T 5 wherein T is as defined above. This general procedure (J) is further illustrated in the following example. 10 Example 720 (General procedure (J)). 9-(3-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HN N NN Cl 3-(2H-Tetrazol-5-yl)-9H-carbazole (example 594, 17 g, 72.26 mmol) was dissolved in N,N dimethylformamide (150 mL). Triphenylmethyl chloride (21.153 g, 75.88 mmol) and triethyl 15 amine (20.14 mL, 14.62 g, 144.50 mmol) were added consecutively. The reaction mixture was stirred for 18 hours at room temperature, poured into water (1.5 L) and stirred for an ad ditional 1 hour. The crude product was filtered off and dissolved in dichloromethane (500 mL). The organic phase was washed with water (2 x 250 mL) and dried with magnesium sul fate (1 h). Filtration followed by concentration yielded a solid which was triturated in heptanes 20 (200 mL). Filtration furnished 3-[2-(triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole (31.5 g) which was used without further purification. 'H-NMR (CDCl 3 ): 68.87 (1H, d), 8.28 (1H, bs), 8.22 (1H, dd), 8.13 (1H, d), 7.49 (1H, d), 7.47 7.19 (18H, m); HPLC-MS (Method C): m/z: 243 (triphenylmethyl); Rt = 5.72 min.
WO 2004/080480 PCT/DK2004/000158 297 3-[2-(Triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole (200 mg, 0.42 mmol) was dissolved in methyl sulfoxide (1.5 mL). Sodium hydride (34 mg, 60 %, 0.85 mmol) was added, and the resulting suspension was stirred for 30 min at room temperature. 3-Chlorobenzyl chloride (85 pL, 108 mg, 0.67 mmol) was added, and the stirring was continued at 40 oC for 18 hours. 5 The reaction mixture was cooled to ambient temperature and poured into 0.1 N hydrochloric acid (aq.) (15 mL). The precipitated solid was filtered off and washed with water (3 x 10 rnL) to furnish 9-(3-chlorobenzyl)-3-[2-(triphenylmethyl)-2H-tetrazol-5-yl]-9H-carbazole, which was dissolved in a mixture of tetrahydrofuran and 6 N hydrochloric acid (aq.) (9:1) (10 mL) arid stirred at room temperature for 18 hours. The reaction mixture was poured into water (100 10 mL). The solid was filtered off and rinsed with water (3 x 10 mL) and dichloromethane (3 x 10 mL) to yield the title compound (127 mg). No further purification was necessary. 1 H-NMR (DMSO-d 6 ): 38.89 (1H, d), 8.29 (1H, d), 8.12 (1H, dd), 7.90 (1H, d), 7.72 (1H, d), 7.53 (1H, t), 7.36-7.27 (4H, m), 7.08 (1H, bt), 5.78 (2H, s); HPLC-MS (Method B): m/z: 360 (M+1); Rt = 5.07 min. 15 The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization from e.g. aqueous sodium hydroxide (1 N) or by chromatography. 20 Example 721 (General Procedure (J)). 9-(4-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N N N N CI HPLC-MS (Method C): m/z: 360 (M+1); Rt = 4.31 min. 25 Example 722 (General Procedure (J)). 9-(4-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N N N ' / CH 3 WO 2004/080480 PCT/DK2004/000158 298 HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.26 min. Example 723 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(4-trifluoromethylbenzyl)-9H-carbazole HN N 5 / CF 3 HPLC-MS (Method C): m/z: 394 (M+1); Rt = 4.40 min. Example 724 (General Procedure (J)). 9-(4-Benzyloxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N= HNN. NN 10 HPLC-MS (Method C): m/z: 432 (M+1); Rt = 4.70 min. Example 725 (General Procedure (J)). 9-(3-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N "N 15
CH
3 HPLC-MS (Method C): mlz: 340 (M+1); Rt = 4.25 min. Example 726 (General Procedure (J)). 9-Benzyl-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 299 ,N:=N HN'N.N N 'H-NMR (DMSO-d 6 ): 88.91 (1H, dd), 8.30 (1H, d), 8.13 (1H, dd), 7.90 (1H, d), 7.73 (1H, d), 7.53 (1H, t), 7.36-7.20 (6H, m), 5.77 (2H, s). 5 Example 727 (General Procedure (J)). 9-(4-Phenylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N= HN 'H-NMR (DMSO-d 6 ): 38.94 (1H, s), 8.33 (1H, d), 8.17 (1H, dd), 7.95 (1H, d), 7.77 (1H, d), 7.61-7.27 (1 1H, m), 5.82 (2H, s). 10 Example 728 (General Procedure (J)). 9-(3-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
,N=N
N N N O-CH3 HPLC-MS (Method C): m/z: 356 (M+1); Rt = 3.99 min. 15 Example 729 (General Procedure (J)). 9-(Naphthalen-2-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 300 HI' N N= N NN HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.48 min. Example 730 (General Procedure (J)). 5 9-(3-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N N Br HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.33 min. Example 731 (General Procedure (J)). 10 9-(Biphenyl-2-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HN N HPLC-MS (Method C): m/z: 402 (M+1); Rt = 4.80 min. Example 732 (General Procedure (J)). 15 3-(2H-Tetrazol-5-yl)-9-[4-(1,2,3-thiadiazol-4-yl)benzyl]-9H-carbazole WO 2004/080480 PCT/DK2004/000158 301 INz N Example 733 (General Procedure (J)). 9-(2'-Cyanobiphenyl-4-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,NF zN N HN N 5 N 'H-NMR (DMSO-d 6 ): .8.91 (1H, d), 8.31 (1H, d), 8.13 (1H, dd), 7.95 (1H, d), 7.92 (1H, d), 7.78 (1H, d), 7.75 (1H, dt), 7.60-7.47 (5H, m), 7.38-7.28 (3H, m), 5.86 (2H, s); HPLC-MS (Method C): m/z: 427 (M+1); Rt = 4.38 min. 10 Example 734 (General Procedure (J)). 9-(4-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HN NN HPLC-MS (Method C): m/z: 452 (M+1); Rt = 4.37 min. 15 Example 735 (General Procedure (J)). 9-(3,5-Bis(trifluoromethyl)benzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 302 HN N N SN
CF
3
CF
3 HPLC-MS (Method C): m/z: 462 (M+1); Rt = 4.70 min. Example 736 (General Procedure (J)). 5 9-(4-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,N=N HN N \/ Br 1 H-NMR (DMSO-d 6 ): 68.89 (1H, d), 8.29 (1H, d), 8.11 (1H, dd), 7.88 (1H, d), 7.70 (1H, d), 7.52 (1H, t), 7.49 (2H, d), 7.31 (1H, t), 7.14 (2H, d), 5.74 (2H, s); HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.40 min. 10 Example 737 (General Procedure (J)). 9-(Anthracen-9-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,N HN N HPLC-MS (Method C): m/z: 426 (M+1); Rt = 4.78 min. 15 Example 738 (General Procedure (J)). 9-(4-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 303 HN
N
HN N OH 3.6 fold excess sodium hydride was used. 1 H-NMR (DMSO-d 6 ): 812.89 (1H, bs), 8.89 (1H, d), 8.30 (1H, d), 8.10 (1H, dd), 7.87 (1H, d), 5 7.86 (2H, d), 7.68 (1H, d), 7.51 (1H, t), 7.32 (1H, t), 7.27 (2H, d), 5.84 (2H, s); HPLC-MS (Method C): m/z: 370 (M+1I); Rt = 3.37 min. Alternative mode of preparation of 9-(4-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H carbazole: 10 Carbazole (52.26 g, 0.30 mol) was dissolved in dichloromethane (3 L) and silicagel (60 mesh, 600 g) was added to the mixture and the mixture was cooled to 10 °C. A mixture of N-bromosuccinimide (NBS, 55 g, 0.30 mol) in dichloromethane (400 mL) was added at 10 'C. After addition, the mixture was allowed to reach room temperature. After standing for 42 15 hours, the mixture was filtered, and the solid was washed with dichloromethane (4 x 200 mL), the combined filtrates were washed with water (300 mL) and dried over Na 2
SO
4 . Evapora tion in vacuo to dryness afforded 77 g of crude product. Recrystallization from 2-propanol (800 mL) afforded 71 % 3-bromocarbazole. 20 To a stirred solution of 3-bromocarbazole (63 g, 0.256 mol) in N-methylpyrrolidone (900 mL) was added cuprous cyanide (CuCN, 25.22 g, 0.28 mol) and the mixture was heated to 190 *C. After 9 hours of heating, the mixture was cooled to room temperature. The mixture was concentrated by bulb-to-bulb distillation (100 OC, 0.1 mm Hg). The residue was treated with NH 4 OH (25%, 300 mL) and subsequently extracted with ethyl acetate (10%) in toluene. 25 The organic layer was dried over Na 2
SO
4 and concentrated by bulb-to-bulb distillation (100 oC, 0.1 mm Hg) to give 34 g (70%) of 3-cyanocarbazole. Sodium hydride 55-60% in mineral oil (3.7 g, 0.093 mol) was added in portions to a stirred, cooled (5 °C) mixture of 3-cyanocarbazole (17.5 g, 0.091 mol) in N,N 30 dimethylformamide (200 mL). After 0.5 hours, a solution of methyl 4-bromomethylbenzoate (22.9 g, 100 mmol) in N,N-dimethylformamide (80 mL) was added dropwise to the cooled WO 2004/080480 PCT/DK2004/000158 304 mixture. The mixture was subsequently slowly warmed to room temperature and stirred overnight. The mixture was poured into ice water and extracted with dichloromethane (2 x 200 mL), the organic layer was washed several times with water, dried over Na 2
SO
4 and concentrated in vacuo. A mixture of ethyl acetate and heptane (1/1, 50 mL) was added to the 5 concentrate and the solid was product filtered off. Yield 24 g (78%) of 4-(3-cyanocarbazol-9 ylrnethyl)benzoic acid methyl ester. Sodium azide (7.8 g, 0.12 mol) and ammonium chloride (6.42 g , 0.12 mol) were added to a stirred mixture of 4-(3-cyanocarbazol-9-ylmethyl)benzoic acid methyl ester (24.8 10 g, 0.073 mol) in N,N-dimethylformamide (130 mL) and the mixture was heated to 110 oC. Af ter 48 hours, the mixture was cooled to room temperature and poured into water (500 mL) and cooled to 5 "C. Hydrochloric acid (10 N) was then added to pH = 2. After stirring for 1 hour at 5 OC the precipitate was filtered off and washed with water. The solid obtained was air dried. Yield 27.9 g of 4-[3-(1H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester. 15 31.1 g of 4-[3-(1H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester was added to a solution of sodium hydroxide (8.76 g, 0.219 mol) in water (150 mL) and the mixture was heated to 80 OC, after 0.5 h activated carbon (0.5 g) was added and the mixture was filtered through celite. The filtrate was treated with hydrochloric acid (10 N) to pH = 1 and the formed 20 precipitate was filtered off and air dried. This procedure was repeated as the first treatment did not give complete hydrolysis of the ester. Finally the product was dissolved in 2-propanol, the filtered the mother liquor was concentrated to approximately 100 mL and the product was isolated by filtration to afford 19 g of the title compound. After evaporation of the mother liq uor to dryness and re-treatment with 2-propanol further 8 g of product was isolated resulting 25 in a yield of 90 %. Example 739 (General Procedure (J)). 9-(2-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HN NN CI 30 HPLC-MS (Method B): m/z: 360 (M+1); Rt = 5.30 min.
WO 2004/080480 PCT/DK2004/000158 305 Example 740 (General Procedure (J)). 9-(4-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,NzN HN N N \ F 5 'H-NMR (DMSO-d): 38.88 (1H, d), 8.28 (1H, d), 8.10 (1H, dd), 7.89 (1H, d), 7.72 (1H, d), 7.52 (1H, t), 7.31 (1H, t), 7.31-7.08 (41-1, m), 5.74 (2H, s); HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.10 min. Example 741 (General Procedure (J)). 10 9-(3-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HN. N F 1 H-NMR (DMSO-d): 68.89 (1H, d), 8.29 (1H, d), 8.12 (1H, dd), 7.90 (1H, d), 7.72 (1H, d), 7.53 (1H, t), 7.37-7.27 (2H, m), 7.12-7.02 (2H, m), 6.97 (1H, d), 5.78 (2H, s); HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.10 min. 15 Example 742 (General Procedure (J)). 9-(2-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HN N NN HPLC-MS (Method C): m/z: 452 (M+1); Rt = 4.58 min. 20 Example 743 (General Procedure (J)). 9-(3-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 306 N HN N OH O 3.6 fold excess sodium hydride was used. 1 H-NMR (DMSO-de): 812.97 (1H, bs), 8.90 (1H, bs), 8.30 (1H, d), 8.12 (1H, bd), 7.89 (1H, d), 5 7.82 (1 H, m), 7.77 (1H, bs), 7.71 (1H, d), 7.53 (1 H, t), 7.46-7.41 (2H, m), 7.32 (1H, t), 5.84 (2H, s); HPLC-MS (Method C): m/z: 370 (M+1); Rt = 3.35 min. Example 744 (General Procedure (J)). 9-[4-(2-Propyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole HN ,N-N HN'N @ N O CH 3 10
CH
3 1 H-NMR (DMSO-de): 68.87 (1H, d), 8.27 (1H, d), 8.10 (1H, dd), 7.87 (1H, d), 7.71 (1H, d), 7.51 (1H, t), 7.31 (1H, t), 7.15 (2H, d), 7.12 (2H, d), 5.69 (2H, s), 2.80 (1H, sept), 1.12 (6H, d); HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.73 min. 15 Example 745 (General Procedure (J)). 9-(3,5-Dimethoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,N= HN. N N N O N CH3 3 OCHz3 HPLC-MS (Method C): m/z: 386 (M+1); Rt = 4.03 min.
WO 2004/080480 PCT/DK2004/000158 307 Example 746 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(2,4,5-trifluorobenzyl)-9H-carbazole N HN NF F F HPLC-MS (Method B): m/z: 380 (M+1); Rt = 5.00 min. 5 Example 747 (General Procedure (J)). N-Methyl-N-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide N HN N
H
3c " N HPLC-MS (Method B): m/z: 383 (M+1); Rt = 4.30 min. 10 Example 748 (General Procedure (J)). 9-(4-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HN, N HN
CH
3 1 H-NMR (DMSO-d6): 88.86 (1H, d), 8.26 (1H, d), 8.10 (1H, dd), 7.90 (1H, d), 7.73 (1H, d), 15 7.51 (1H, t), 7.30 (1H, t), 7.18 (2H, d), 6.84 (2H, d), 5.66 (2H, s), 3.67 (3H, s); HPLC-MS (Method B): m/z: 356 (M+1); Rt = 4.73 min. Example 749 (General Procedure (J)). 9-(2-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 308
HN.
N N
ON
3 1 H-NMR (DMSO-d 6 ): 68.87 (1H, d), 8.27 (1H, d), 8.09 (1H, dd), 7.77 (1H, d), 7.60 (1H, d), 7.49 (1H, t), 7.29 (1H, t), 7.23 (1H, bt), 7.07 (1H, bd), 6.74 (1H, bt), 6.61 (1H, bd), 5.65 (2H, s), 3.88 (3H, s); HPLC-MS (Method B): m/z: 356 (M+1); Rt = 4.97 min. 5 Example 750 (General Procedure (J)). 9-(4-Cyanobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HN'NN HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.74 min. 10 Example 751 (General Procedure (J)). 9-(3-Cyanobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HN. NN N HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.73 min. 15 Example 752 (General Procedure (J)). 9-(5-Chloro-2-methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 309 N HN . HN NCl ,_o 0
CH
3 1 H-NMR (DMSO-d 6 ): 88.87 (1H, d), 8.35 (1H, d), 8.10 (1H, dd), 7.73 (1H, d), 7.59 (1H, d), 7.49 (1H, t), 7.29 (1H, t), 7.27 (1H, dd), 7.11 (1H, d), 6.51 (1H, d), 5.63 (2H, s), 3.88 (3H, s); HPLC-MS (Method C): m/z: 390 (M+1); Rt = 4.37 min. 5 Example 753 (General Procedure (J)). N-Phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide N= HN HN N N 1 H-NMR (DMSO-d 6 ): 810.54 (1H, s), 8.87 (1H, bs), 8.27 (1H, d), 8.12 (1H, bd), 7.83 (1H, d), 10 7.66 (1H, d), 7.61 (2H, d), 7.53 (1H,t), 7.32 (1H, t), 7.32 (2H, t), 7.07 (1H, t), 5.36 (2H, s); HPLC-MS (Method C): m/z: 369 (M+1); Rt = 3.44 min. Example 754 (General Procedure (J)). N-Butyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide HN. N N0
H
15
CH
3 1 H-NMR (DMSO-d 6 ): 58.85 (1H, d), 8.31 (1H, t), 8.25 (1H, d), 8.10 (1H, dd), 7.75 (1H, d), 7.58 (1H, d), 7.52 (1H, t), 7.30 (1H, t), 5.09 (2H, s), 3.11 (2H, q), 1.42 (2H, quint), 1.30 (2H, sext), 0.87 (3H, t); HPLC-MS (Method C): m/z: 349 (M+1); Rt = 3.20 min.
WO 2004/080480 PCT/DK2004/000158 310 Example 755 (General Procedure (J)). 9 -(2,4-Dichlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole PJNN ,N= N N /l C I CCl 1 H-NMR (DMSO-d): 88.92 (1H, d), 8.32 (1H, d), 8.09 (1H, dd), 7.76 (1H, d), 7.74 (1H, d), 5 7.58 (1H, d), 7.51 (1H, t), 7.33 (1H, t), 7.23 (1H, dd), 6.42 (1H, d), 5.80 (2H, s); HPLC-MS (Method B): m/z: 394 (M+1); Rt = 5.87 min. Example 756 (General Procedure (J)). 9-(2-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
HN._-
N N " 10
H
3 C 1 H-NMR (DMSO-d 6 ): 88.92 (1H, d), 8.32 (1H, d), 8.08 (1H, dd), 7.72 (1H, d), 7.55 (1H, d), 7.48 (1H, t), 7.32 (1H, t), 7.26 (1H, d), 7.12 (1H, t), 6.92 (1H, t), 6.17 (1H, d), 5.73 (2H, s), 2.46 (3H, s); HPLC-MS (Method B): m/z: 340 (M+1); Rt = 5.30 min. 15 Example 757 (General Procedure (J)). 9-(3-Nitrobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,NzN N
NO
2 HPLC-MS (Method C): m/z: 371 (M+1); Rt = 3.78 min. 20 Example 758 (General Procedure (J)). 9-(3,4-Dichlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 311
N
HN N Cl HPLC-MS (Method B): m/z: 394 (M+1); Rt = 5.62 min. Example 759 (General Procedure (J)). 5 9-(2,4-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N N N ' F F 1 H-NMR (DMSO-d 6 ): 58.89 (1H, d), 8.29 (1H, d), 8.11 (1H, dd), 7.88 (1H, d), 7.69 (1H, d), 7.52 (1H, t), 7.36-7.24 (2H, m), 7.06-6.91 (2H, m), 5.78 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min. 10 Example 760 (General Procedure (J)). 9-(3,5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N HNN F F 1 H-NMR (DMSO-d): 68.90 (1H, bs), 8.31 (1H, d), 8.13 (1H, bd), 7.90 (1H, d), 7.73 (1H, d), 15 7.54 (1H, t), 7.34 (1H, t), 7.14 (1H, t), 6.87 (2H, bd), 5.80 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min. Example 761 (General Procedure (J)). 9-(3,4-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 312 HNN N F F 1 H-NMR (DMSO-d 6 ): 88.89 (11H, bs), 8.29 (1H, d), 8.12 (1H, bd), 7.92 (1H, d), 7.74 (1H, d), 7.54 (1 H, t), 7.42-7.25 (3H, m), 6.97 (1H, bm), 5.75 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min. 5 Example 762 (General Procedure (J)). 9-(3-lodobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N=N HN.N NN HPLC-MS (Method B): m/z: 452 (M+1); Rt = 5.50 min. 10 Example 763 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[3-(trifluoromethyl)benzyl]-9H-carbazole
HN.
N N
CF
3 'H-NMR (DMSO-d 6 ): 88.89 (1H, d), 8.30 (1H, d), 8.11 (1H, dd), 7.90 (1H, d), 7.72 (1H, d), 15 7.67 (1H, bs), 7.62 (1H, bd), 7.53 (1H, t), 7.50 (1H, bt), 7.33 (1H, bd), 7.32 (1H, t), 5.87 (2H, s); HPLC-MS (Method B): m/z: 394 (M+1); Rt = 5.40 min. Example 764 (General Procedure (J)). N-(4-Carboxyphenyl)-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide WO 2004/080480 PCT/DK2004/000158 313
N-
N N HNN N N OH 3.6 fold excess sodium hydride was used. 5 HPLC-MS (Method B): m/z: 413 (M+1); Rt = 3.92 min. Example 765 (General Procedure (J)). N-(2-Propyl)-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide N
CH
3 10 HPLC-MS (Method B): m/z: 335 (M+1); Rt = 3.70 min. Example 766 (General Procedure (J)). N-Benzyl-N-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide ,' N NN HN.NN 0 15 HPLC-MS (Method B): m/z: 459 (M+1); Rt = 5.37 min. Example 767 (General Procedure (J)). N-[4-(2-Methyl-2-propyl)phenyl]-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide WO 2004/080480 PCT/DK2004/000158 314 HN N HN N Wo N Me Me Me HPLC-MS (Method B): m/z: 425 (M+1); Rt = 5.35 min. Example 768 (General Procedure (J)). 5 N-Phenethyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide FIN. N= HN N H HPLC-MS (Method C): m/z: 397 (M+1); Rt = 3.43 min. Example 769 (General Procedure (J)). 10 3-(2H-Tetrazol-5-yl)-9-[2-(trifluoromethyl)benzyl]-9H-carbazole I N HN. N
F
3 C HPLC-MS (Method C): m/z: 394 (M+1); Rt = 4.44 min. Example 770 (General Procedure (J)). 15 9-[2-Fluoro-6-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 315 N= HNN NN
F
3 C HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.21 min. Example 771 (General Procedure (J)). 5 9-[2,4-Bis(trifluoromethyl)benzyl)]-3-(2H-tetrazol-5-yl)-9H-carbazole ,NzN HN
CF
3
F
3 C HPLC-MS (Method C): m/z: 462 (M+1); Rt = 4.82 min. Example 772 (General Procedure (J)). 10 3-(2H-Tetrazol-5-yl)-9-(2,4,6-trimethylbenzyl)-9H-carbazole N=N HN. / CH 3
H
3 C HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.59 min. Example 773 (General Procedure (J)). 15 9-(2,3,5,6-Tetramethylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,N=N HN NC N " NC
H
3 C 3 11 3 C
CH
3 WO 2004/080480 PCT/DK2004/000158 316 HPLC-MS (Method C): m/z: 382 (M+1); Rt = 4.47 min. Example 774 (General Procedure (J)). 9-[(Naphthalen-1-yl)methyl]-3-(2H-tetrazol-5-yl)-9H-carbazole N HNAN 5 HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.43 min. Example 775 (General Procedure (J)). 9-[Bis(4-fluorophenyl)methyl]-3-(2H-tetrazol-5-yl)-9H-carbazole N HN /F 10 F HPLC-MS (Method C): m/z: 438 (M+1); Rt = 4.60 min. Example 776 (General Procedure (J)). 9-(2-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HN. _: NN 15 Br HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.50 min. Example 777 (General Procedure (J)). 9-(2-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 317 N N N F HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.09 min. Example 778 (General Procedure (J)). 5 9-(4-Carboxy-2-methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole ,N N N= HN N OH
H
3 C O In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): mlz: 384 (M+1); Rt = 3.56 min. 10 Example 779 (General Procedure (J)). 9-(2-Phenylethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole H N N= HN N HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.08 min. 15 Example 780 (General Procedure (J)). 9-[2-Fluoro-5-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole WO 2004/080480 PCT/DK2004/000158 318
HNN
N
HN"NNCF
3 F HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.34 min. Example 781 (General Procedure (J)). 5 9-(4-Carboxy-2-fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N= HN . NN OH F O 3-Fluoro-4-methylbenzoic acid (3.0 g, 19.5 mmol) and benzoyl peroxide (0.18 g, 0.74 mmol) were suspended in benzene. The mixture was purged with N 2 and heated to reflux. N Bromosuccinimide (3.47 g, 19.5 mmol) was added portionwise, and reflux was maintained for 10 18 hours. The reaction mixture was concentrated, and the residue was washed with water (20 mL) at 70 'C for 1 hour. The crude product was isolated by filtration and washed with ad ditional water (2 x 10 mL). The dry product was recrystallized from heptanes. Filtration fur nished 4-bromomethyl-3-fluorobenzoic acid (1.92 g) which was used in the following step ac cording to General Procedure (J). 15 In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 388 (M+1); Rt = 3.49 min. Example 782 (General Procedure (J)). 5-{4-[[(3-(2H-Tetrazol-5-yl)carbazol-9-yl)methyl]naphthalen-1-yl]oxy}pentanoic Acid N 0. HN / O H 20 5-[(4-Formylnaphthalen-1-yl)oxy]pentanoic acid intermediate obtained in example 470(3.0 g, 11.0 mmol) was dissolved in a mixture of methanol and tetrahydrofuran (9:1) (100 mL), and WO 2004/080480 PCT/DK2004/000158 319 sodium borohydride (1.67 g, 44.1 mmol) was added portionwise at ambient temperature. Af ter 30 minutes, the reaction mixture was concentrated to 50 mL and added to hydrochloric acid (0.1 N, 500 mL). Additional hydrochloric acid (1 N, 40 mL) was added, and 5-[(4 hydroxymethyl-naphthalen-1 -yl)oxy]pentanoic acid (2.90 g) was collected by filtration. To the 5 crude product was added concentrated hydrochloric acid (100 mL), and the suspension was stirred vigorously for 48 hours at room temperature. The crude product was filtered off and washed with water, until the pH was essentially neutral. The material was washed with hep tanes to furnish 5-[(4-chloromethylnaphthalen-1-yl)oxy]pentanoic acid (3.0 g) which was used in the following step according to General Procedure (J). 10 In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): mlz: 492 (M+1); Rt = 4.27 min. Example 783 (General procedure (J)) 15 9-(2,3-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole HNN N F F HPLC-MS (Method C): m/z= 362 (M+1); Rt = 4.13 min. Example 784 (General procedure (J)) 20 9-(2,5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N0 N F- d F3 HPLC-MS (Method C): mlz = 362 (M+1); Rt = 4.08 min.
WO 2004/080480 PCT/DK2004/000158 320 Example 785 (General procedure (J)) 9-Pentafluorophenylmethyl-3-(2H-tetrazol-5-yl)-9H-carbazole HN -N NN F N F F HPLC-MS (Method C): m/z = 416 (M+1); Rt = 4.32 min. 5 Example 786 (General procedure (J)) 9-(2,6-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole N N N F N N F HPLC-MS (Method C): m/z = 362 (M+1); Rt = 3.77 min. 10 Further compounds of the invention that may be prepared according to general procedure (J), and includes: WO 2004/080480 PCT/DK2004/000158 321 Example 787 Example 788 Example 789 NzN NNN __ HN HN. HN N N N N' N S ,. N ' N " N F Br F 3 C Example 790 Example 791 Example 792 HN.- NN..N N NN N N N, H N N' H NN I F N Me FC Me Example 793 Example 794 Example 795 N NzN NN HNN H N N HN.N F F F C(DJ OH Example 796 Example 797 Example 798 HNN H NN H N HN NNH FF F Example 799 H. -z N The following compounds of the invention may be prepared eg. from 9-(4-bromobenzyl)-3 (2H-tetrazol-5-yl)-9H-carbazole (example 736) or from 9-(3-bromobenzyl)-3-(2H-tetrazol-5 yl)-9H-carbazole (example 730) and aryl boronic acids via the Suzuki coupling reaction eg as 5 described in Littke, Dai & Fu J. Am. Chem. Soc., 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-tert-butylphosphine)palladium (0).
WO 2004/080480 PCT/DK2004/000158 322 Example 800 Example 801 Example 802 TNClNHC N N N HN\1H HNH OH 0 Example 803 Example 804 Example 805 Exm 0 General procedure (K))frpeaainofcmonso.enrlfrua10 H 1-N H- -N \H NiNl N NN N -N NN 0 General procedure (K) for preparation of compounds of general formula 110: N N NaN 3 HN T CI N 4 CI l~ ~ ____ I \ LiCI N N N H NaH H 110 H 5 wherein T is as defined above. The general procedure (K) is further illustrated by the following example: Example 806 (General procedure (K)). 10 1 -Benzyl-5-(2H-tetrazol-5-yl)-1 H-indole MN HN NN 5-Cyanoindole (1.0 g, 7.0 mmol) was dissolved in N,N-dimethylformamide (14 mL) and cooled in an ice-water bath. Sodium hydride (0.31 g, 60 %, 7.8 mmol) was added, and the resulting suspension was stirred for 30 min. Benzyl chloride (0.85 mL, 0.94 g, 7.4 mmol) was WO 2004/080480 PCT/DK2004/000158 323 added, and the cooling was discontinued. The stirring was continued for 65 hours at room temperature. Water (150 mL) was added, and the mixture was extracted with ethyl acetate (3 x 25 mL). The combined organic phases were washed with brine (30 mL) and dried with so dium sulfate (1 hour). Filtration and concentration yielded the crude material. Purification by 5 flash chromatography on silica gel eluting with ethyl acetate/heptanes = 1:3 afforded 1.60 g 1-benzyl-1 H-indole-5-carbonitrile. HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.17 min. 10 1-Benzyl-1H-indole-5-carbonitrile was transformed into 1-benzyl-5-(2H-tetrazol-5-yl)-1 H indole by the method described in general procedure (J) and in example 594. Purification was done by flash chromatography on silica gel eluting with dichloromethane/methanol = 9:1. HPLC-MS (Method C): m/z: 276 (M+1); Rt = 3.35 min. 15 The compounds in the following examples were prepared by the same procedure. Example 807 (General procedure (K)). 1-(4-Bromobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole HN. NK 20/ Br 20 HPLC-MS (Method C): m/z: 354 (M+1); Rt = 3.80 min. Example 808 (General procedure (K)). 1-(4-Phenylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole H N NN 25 1 H-NMR (200 MHz, DMSO-d): 6 = 5.52 (2H, s), 6.70 (1H, d), 7.3-7.45 (6H, m), 7.6 (4H, m), 7.7-7.8 (2H, m), 7.85(1 H, dd), 8.35 (1 H, d).
WO 2004/080480 PCT/DK2004/000158 324 Calculated for C 22
H
17 Ns, H 2 0: 73.32% C; 5.03% H; 19.43% N. Found: 73.81% C; 4.90% H; 19.31% N. 5 Example 809 4'-[5-(2H-Tetrazol-5-yl)indol-1-ylmethyl]biphenyl-4-carboxylic acid N. HN N N /. OH O 0 5-(2H-Tetrazol-5-yl)-1 H-indole (Syncom BV, Groningen, NL) (1.66g, 8.9 mmol) was treated with trityl chloride (2.5 g, 8.9 mmol) and triethyl amine (2.5 mL, 17.9 mmol) in DMF(25 mL) by 10 stirring at RT overnight. The resulting mixture was treated with water. The gel was isolated, dissolved in methanol, treated with activated carbon; filtered and evaporated to dryness in vacuo. This afforded 3.6 g (94%) of crude 5-(2-trityl-2H-tetrazol-5-yl)-1H-indole. HPLC-MS (Method C): m/z = 450 (M+23); Rt. = 5.32 min. 15 4-Methylphenylbenzoic acid (5 g, 23.5 mmol) was mixed with CCl 4 (100 mL) and under an atmosphere of nitrogen, the slurry was added N-Bromosuccinimide (4.19 g, 23.55 mmol) and dibenzoyl peroxide (0.228 g, 0.94 mmol). The mixture was subsequently heated to reflux for 0.5 hour. After cooling, DCM and water (each 30 mL) were added. The resulting precipitate 20 was isolated, washed with water and a small amount of methanol. The solid was dried in vacuo to afford 5.27 g (77%) of 4'-bromomethylbiphenyl-4-carboxylic acid. HPLC-MS (Method C): m/z = 291 (M+1); Rt. = 3.96 min. 25 5-(2-Trityl-2H-tetrazol-5-yl)-1H-indole (3.6 g, 8.4 mmol) was dissolved in DMF (100 mL). Un der nitrogen, NaH (60 % suspension in mineral oil, 34 mmol) was added slowly. 4' Bromomethylbiphenyl-4-carboxylic acid (2.7 g, 9.2 mmol) was added over 5 minutes and the WO 2004/080480 PCT/DK2004/000158 325 resulting slurry was heated at 40 °C for 16 hours. The mixture was poured into water (1 00mL) and the precipitate was isolated by filtration and treated with THF/6N HCI (9/1) (70 mL) at room temperature for 16 hours. The mixture was subsequently evaporated to dryness in vacuo, the residue was treated with water and the solid was isolated by filtration and 5 washed thoroughly 3 times with DCM. The solid was dissolved in hot THF (400 mL) treated with activated carbon and filtered. The filtrate was evaporated in vacuo to dryness. This af forded 1.6 g (50%) of the title compound. HPLC-MS (Method C): m/z = 396 (M+1); Rt. = 3.51 min. 10 Example 810 (General procedure (K)). 5-(2H-Tetrazol-5-yl)-1 H-indole SNN HNN NN H 5-(2H-Tetrazol-5-yl)-IH-indole was prepared from 5-cyanoindole according to the method 15 described in example 594. HPLC-MS (Method C): m/z: 186 (M+1); Rt= 1.68 min. Example 811 (General procedure (K)). 20 1 -Benzyl-4-(2H-tetrazol-5-yl)-1H-indole H N-N I \ NN 1-Benzyl-1 H-indole-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 806. HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.24 min. 25 1 -Benzyl-4-(2H-tetrazol-5-yl)-1 H-indole was prepared from 1-benzyl-1H-indole-4-carbonitrile according to the method described in example 594.
WO 2004/080480 PCT/DK2004/000158 326 HPLC-MS (Method C): m/z: 276 (M+1); Rt = 3.44 min. General procedure (L) for preparation of compounds of general formula l,: ,N N
,N-
N NN~ HN N ' Pal-Cl ! DIPEA= Poa-N N i:LiHMDS I DMSO DMF / DC P ii: X-CH 2 -T-H; X= CI, Br H H ,N N ,N -- N Pol-N HN N 0.1 N HCI in N '"N THF / Et 2 0 / EtOH = 8:1:1 N 5 i H wherein T is as defined above and Pol- is a polystyrene resin loaded with a 2-chlorotrityl linker, graphically shown below: I Pol- = PS 10 This general procedure (L) is further illustrated by the following example: Example 812 (General procedure (L)). 5-(2H-Tetrazol-5-yl)-1 -[3-(trifluoromethyl)benzyll-1 H-indole N'N HN.N 15
CF
3 2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloro methane (2 mL) for 30 min. The solvent was drained, and a solution of 5-(2H-tetrazol-5-yl)- WO 2004/080480 PCT/DK2004/000158 327 1H-indole (example 810) (63 mg, 0.34 mmol) in a mixture of N,N-dimethylformamide, di chloromethane and N,N-di(2-propyl)ethylamine (DIPEA) (5:5:2) (1.1 mL) was added. The re action mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed consecutively with N,N-dimethylformamide (2 x 4 mL), 5 dichloromethane (6 x 4 mL) and methyl sulfoxide (2 x 4 mL). Methyl sulfoxide (1 mL) was added, followed by the addition of a solution of lithium bis(trimethylsilyl)amide in tetrahydrofu ran (1.0 M, 0.57 mL, 0.57 mmol). The mixture was shaken for 30 min at room temperature, before 3-(trifluoromethyl)benzyl bromide (273 mg, 1.14 mmol) was added as a solution in methyl sulfoxide (0.2 mL). The reaction mixture was shaken for 20 hours at room tempera 10 ture. The drained resin was washed consecutively with methyl sulfoxide (2 x 4 mL), di chloromethane (2 x 4 mL), methanol (2 x 4 mL), dichloromethane (2 x 4 mL) and tetrahydro furan (4 mL). The resin was treated with a solution of hydrogen chloride in tetrahydrofuran, ethyl ether and ethanol = 8:1:1 (0.1 M, 3 mL) for 6 hours at room temperature. The resin was drained and the filtrate was concentrated in vacuo. The crude product was re-suspended in 15 dichloromethane (1.5 mL) and concentrated three times to afford the title compound (35 mg). No further purification was necessary. HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.35 min. 1 H-NMR (DMSO-d 6 ): 38.29 (1H, s), 7.80 (1H, dd), 7.72 (2H, m), 7.64 (2H, bs), 7.56 (1H, t), 20 7.48 (1H, d), 6.70 (1H, d), 5.62 (2H, s). The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization or by chromatography. 25 Example 813 (General procedure (L)). 1-(4-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole NN HN.N N K ' /Cl HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.11 min. 30 Example 814 (General procedure (L)). 1-(2-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole WO 2004/080480 PCT/DK2004/000158 328 ,N N HN N. IN CI HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.05 min. Example 815 (General procedure (L)). 5 1-(4-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole ,N HNN NN
CH
3 HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.68 min. Example 816 (General procedure (L)). 10 1-(4-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole ,N NN HNN N \ /CH 3 HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min. Example 817 (General procedure (L)). 15 5-(2H-Tetrazol-5-yl)-1l-[4-(trifluoromethyl)benzyl]-1 H-indole ,N'"N HNN .\ /
CF
3 HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.18 min.
WO 2004/080480 PCT/DK2004/000158 329 Example 818 (General procedure (L)). 1-(3-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole Ht N::! ,NN Cl HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.01 min. 5 Example 819 (General procedure (L)). 1-(3-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole N N HNN N
CH
3 HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min. 10 Example 820 (General procedure (L)). 1-(2,4-Dichlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole ,N N HN.. N / CI Cl HPLC-MS (Method B): mlz: 344 (M+1); Rt = 4.41 min. 15 Example 821 (General procedure (L)). 1-(3-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole WO 2004/080480 PCT/DK2004/000158 330 HN N N
O-CH
3 HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.64 min. Example 822 (General procedure (L)). 5 1-(4-Fluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole ' N HN. N AN \-- F HPLC-MS (Method B): m/z: 294 (M+1); Rt = 3.71 min. Example 823 (General procedure (L)). 10 1-(3-Fluorobenzyl)-5-(2H-tetrazol-5-yl)- 1H-indole Hl N NN HN NN F HPLC-MS (Method B): m/z: 294 (M+1); Rt = 3.68 min. Example 824 (General procedure (L)). 15 1-(2-lodobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole 11 N NN HPLC-MS (Method B): m/z: 402 (M+1); Rt = 4.11 min.
WO 2004/080480 PCT/DK2004/000158 331 Example 825 (General procedure (L)). 1 -[(Naphthalen-2-yl)methyl]-5-(2H-tetrazol-5-yl)-1 H-indole N NN 5 HPLC-MS (Method B): m/z: 326 (M+1); Rt = 4.18 min. Example 826 (General procedure (L)). 1-(3-Bromobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole ,N N Br 10 HPLC-MS (Method B): m/z: 354 (M+1); Rt = 4.08 min. Example 827 (General procedure (L)). 1-(4-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole HN. N OH 0 O 15 In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 1.7 mL, 1.7 mmol) was used. HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.84 min. Example 828 (General procedure (L)). 20 1-(3-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole WO 2004/080480 PCT/DK2004/000158 332 H N N HN N OH O In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 1.7 mL, 1.7 mmol) was used. HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.91 min. 5 Example 829 (General procedure (L)). 1-(2,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole N N HN NN F F 10 HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.78 min. Example 830 (General procedure (L)). 1-(3,5-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole HN. N
-
F NN 15 F HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.78 min. Example 831 (General procedure (L)). 1-(3,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole WO 2004/080480 PCT/DK2004/000158 333 N N N \ F F HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.81 min. Example 832 (General procedure (L)). 5 1-[4-(2-Propyl)benzyl]-5-(2H-tetrazol-5-yl)-1 H-indole , N N HN.N N
\CH
3
CH
3 HPLC-MS (Method B): m/z: 318 (M+1); Rt = 4.61 min. Example 833 (General procedure (L)). 10 1-(3,5-Dimethoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole HN.. N N HN OCH \_q 3 O-CH3 HPLC-MS (Method B): m/z: 336 (M+1); Rt = 3.68 min. Example 834 (General procedure (L)). 15 1-(2'-Cyanobiphenyl-4-ylmethyl)-5-(2H-tetrazol-5-yl)-I H-indole WO 2004/080480 PCT/DK2004/000158 334 NzN HN N N N HPLC-MS (Method B): m/z: 377 (M+1); Rt = 4.11 min. Example 835 (General procedure (L)). 5 1-(2-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole N NN HNN N
H
3 C HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min. 10 Further compounds of the invention that may be prepared according to general procedure (K) and/or (L) includes: WO 2004/080480 PCT/DK2004/000158 335 Example 836 Example 837 Example 838 ,N N ,N N ,N N HN.N HN Nz HN.N CH, CCH, CF, O-CH, Example 839 Example 840 Example 841
,N=
N
,N=
N ,NzN HN. ' HN.N HNN H4OCH CI F
OCH
3 O-CH Example 842 Example 843 Example 844 HN NN H.NN HN N
N
N
N-
N
N
N NN'N- " HNN HNIN -HNN Example 845 Example 846 Example 847 NP=N ,NN ,NN NNH N.N, H- NN CH, H 3 C Example 848 Example 849 Example 850 Nz N ,Nz N ,N.N HN. -NN HN. N CH, OH 0 Br Example 851 Example 852 Example 853
NN
N ,NN ,NN HN- HN. HN.N F F F F Example 854 Example 855 Example 856 H NzN HN z HN z NN N N
,N
N Ca 0 OH CH
O
WO 2004/080480 PCT/DK2004/000158 336 Example 857 Example 858 Example 859 N-N N-N NN H h N -N , H N N H . N F CF F F The following compounds of the invention may be prepared eg. from 1-(4-bromobenzyl)-5 (2H-tetrazol-5-yl)-1H-indole (example 807) or from the analogue 1-(3-bromobenzyl)-5-(2H tetrazol-5-yl)-IH-indole and aryl boronic acids via the Suzuki coupling reaction eg as de 5 scribed in Littke, Dai & Fu J. Am. Chem. Soc., 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-tert-butylphosphine)palladium (0). Example 860 Example 861 PNN N= HN. N-' HN N OH 0 Example 862 Example 863 Example 864 HNN HN N- HN N N- N N" 10 General procedure (M) for preparation of compounds of general formula 112: NN HN .. N.: N Cl T NaN 3 H N O H
NH
4 CI N SLiCI -N Et3N/DMAP -N H N H H T 1O O12 12 WO 2004/080480 PCT/DK2004/000158 337 wherein T is as defined above. The general procedure (M) is further illustrated by the following example: Example 865 (General procedure (M)). 5 1-Benzoyl-5-(2H-tetrazol-5-yl)-1 H-indole HNN N 0 To a solution of 5-cyanoindole (1.0 g, 7.0 mmol) in dichloromethane (8 mL) was added 4 (dimethylamino)pyridine (0.171 g, 1.4 mmol), triethylamine (1.96 mL, 1.42 g, 14 mmol) and benzoyl chloride (0.89 mL, 1.08 g, 7.7 mmol). The resulting mixture was stirred for 18 hours 10 at room temperature. The mixture was diluted with dichloromethane (80 mL) and washed consecutively with a saturated solution of sodium hydrogencarbonate (40 mL) and brine (40 mL). The organic phase was dried with magnesium sulfate (1 hour). Filtration and concentra tion furnished the crude material which was purified by flash chromatography on silica gel, eluting with ethyl acetate/heptanes = 2:3. 1-Benzoyl-1 H-indole-5-carbonitrile was obtained as 15 a solid. HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4.07 min. 1-Benzoyl-1H-indole-5-carbonitrile was transformed into 1-benzoyl-5-(2H-tetrazol-5-yl)-1H 20 indole by the method described in example 594. HPLC (Method C): Rt = 1.68 min. 25 The compound in the following example was prepared by the same procedure. Example 866 (General procedure (M)). 1-Benzoyl-4-(2H-tetrazol-5-yl)-1 H-indole WO 2004/080480 PCT/DK2004/000158 338 H N-N I 11 N N N 0 1 -Benzoyl-1H-indole-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 865. HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4.24 min. 5 1-Benzoyl-4-(2H-tetrazol-5-yl)-1lH-indole was prepared from 1-benzoyl-1H-indole-4 carbonitrile according to the method described in example 594. HPLC (Method C): Rt = 1.56 min. 10 Example 867 (General procedure (M)) (2-Fluoro-3-trifluoromethylphenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone
,N-
N HNN NN F I F F 0 HPLC-MS (Method B): m/z = 376 (M+1); Rt = 4.32 min. 15 Example 868 (General procedure (M)) (4-Methoxyphenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone HN,. N
CH
3 HPLC-MS (Method B): m/z = 320 (M+1); Rt = 3.70 min. 20 WO 2004/080480 PCT/DK2004/000158 339 Example 869 (General procedure (M)) (3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone HN
,N
O N O 0 HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min. 5 Example 870 (General procedure (M)) (4-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone H , N :N HN N NN O~ N o 0 HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.71 min. 10 Example 871 (General procedure (M)) Naphthalen-2-yl-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone ,N N HN~ NN HPLC-MS (Method C): m/z = 340 (M+1); Rt = 4.25 min. 15 Example 872 (General procedure (M)) (2,3-Difluorophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone WO 2004/080480 PCT/DK2004/000158 340 HN N o 0'F F HPLC-MS (Method B: m/z = 326 (M+1); Rt = 3.85 min. The following known and commercially available compounds do all bind to the His B10 Zn2+ 5 site of the insulin hexamer: Example 873 1-(4-Fluorophenyl)-5-(2H-tetrazol-5-yl)-1 H-indole ,' N
HN--
N N F 10 Example 874 1 -Amino-3-(2H-tetrazol-5-yl)benzene HN.i NH2 15 Example 875 1-Amino-4-(2H-tetrazol-5-yl)benzene ,N-N HN. N
NNH
2 20 WO 2004/080480 PCT/DK2004/000158 341 A mixture of 4-aminobenzonitrile (10 g, 84.6 mmol), sodium azide (16.5 g, 254 mmol) and ammonium chloride (13.6 g, 254 mmol) in DMF was heated at 125 oC for 16 hours. The cooled mixture was filtered and the filtrate was concentrated in vacuo. The residue was added water (200 mL) and diethyl ether (200 mL) which resulted in crystallisation. The mix 5 ture was filtered and the solid was dried in vacuo at 40 OC for 16 hours to afford 5-(4 aminophenyl)-2H-tetrazole. 1 H NMR DMSO-d 6 ): 6 = 5.7 (3H, bs), 6.69 (2H, d), 7.69 (2H, d). HPLC-MS (Method C): mlz: 162 (M+1); Rt = 0,55 min. 10 Example 8761-Nitro-4-(2H-tetrazol-5-yl)benzene N N 0 HNN Nd
I
O Example 8771-Bromo-4-(2H-tetrazol-5-yl)benzene ,NzN HN 15 Br General procedure (N) for solution phase preparation of amides of general formula 113: 0 0 Frag OH + HN 'R Frag 11 N R R' R' 113 20 wherein Frag is any fragment carrying a carboxylic acid group, R is hydrogen, optionally sub stituted aryl or Cs8-alkyl and R' is hydrogen or Cl.
4 -alkyl. Frag-CO 2 H may be prepared eg by general procedure (D) or by other similar procedures de scribed herein, or may be commercially available. 25 The procedure is further illustrated in the following example 878: WO 2004/080480 PCT/DK2004/000158 342 Example 878 (General procedure (N)) N-(4-Chlorobenzyl)-2-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)-1H-indol-1-yl]acetamide 0 o c HNS H HN 0 5 [3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1 -yl]acetic acid (example 478, 90.7 mg, 0.3 mmol) was dissolved in NMP (1 mL) and added to a mixture of 1-ethyl-3-(3-dimethylamino propyl)carbodiimide, hydrochloride (86.4 mg, 0.45 mmol) and 1-hydroxybenzotriazol (68.8 mg, 0.45 mmol) in NMP (1 mL). The resulting mixture was shaken at RT for 2 h. 4 Chlorobenzylamine (51 mg, 0.36 mmol) and DIPEA (46.4 mg, 0.36 mmol) in NMP (1 mL) 10 were added to the mixture and the resulting mixture shaken at RT for 2 days. Subsequently ethyl acetate (10 mL) was added and the resulting mixture washed with 2x10 mL water fol lowed by saturated ammonium chloride (5 mL). The organic phase was evaporated to dry ness giving 75 mg (57%) of the title compound. 15 HPLC-MS (Method C): m/z: 426 (M+1); Rt. = 3.79 min. Example 879 (General procedure (N)) N-(4-Chlorobenzyl)-4-[2-chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide O 0 0 H CI H CI 0 20 HPLC-MS (Method A): m/z: 465 (M+1); Rt = 4.35 min. Example 880 (General procedure (N)) N-(4-Chlorobenzyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide 0 O 25 HPLC-MS (Method A): m/z: 431 (M+0); Rt 3.68 in. 25 HPLC-MS (Method A): mlz: 431 (M+1); Rt = 3.68 min.
WO 2004/080480 PCT/DK2004/000158 343 Example 881 (General procedure (N)) 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]-N-(4-chlorobenzyl)acetamide OO 0 0 HN Br H C1 0 HPLC-MS (Method A): m/z: 483 (M+1); Rt = 4.06 min. 5 Example 882 (General procedure (N)) N-(4-Chlorobenzyl)-2-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetamide 0 N N H O CI o 0 HPLC-MS (Method A): m/z: 403 (M+1); Rt = 4.03 min. 10 Example 883 (General procedure (N)) N-(4-Chlorobenzyl)-3-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]acrylamide 0 0 HN N' H I CI 0 HPLC-MS (Method A): m/z: 399 (M+1); Rt = 3.82. 15 Example 884 (General procedure (N)) N-(4-Chlorobenzyl)-4-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide 0 HN N' 0 "- rN o o HPLC-MS (Method A): m/z: 431 (M+1); Rt = 3.84 min. 20 Example 885 (General procedure (N)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]-N-(4-chlorobenzyl)butyramide 0 HN Br H ,CI O HPLC-MS (Method A): m/z: 511 (M+0); Rt = 4.05 min. H PLC-MS (Method A): mlz: 511 (M+ 1); Rt = 4.05 min.
WO 2004/080480 PCT/DK2004/000158 344 Example 886 (General procedure (N)) 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)-phenoxy]-N-(4-chlorobenzyl) butyramide [Br CI sO 5 0 HPLC-MS (Method A): m/z: 527 (M+1); Rt = 4.77 min. Example 887 (General procedure (N)) N-(4-Chlorobenzyl)-2-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetamide NC O H HN O / 10 HPLC-MS (Method C): m/z: 431 (M+1); Rt. = 4.03 min. Example 888 (General procedure (N)) N-(4-Chlorobenzyl)-3-[3-(2,4-dioxothiazolidin-5-ylidenemethyl)-1H-indol- 1-yl]propionamide O CI N 15 0 HPLC-MS (Method C): m/z: 440 (M+1); Rt. = 3.57 min. Example 889 (General procedure (N)) N-(4-Chlorobenzyl)-4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyramide 0 K0 ~o HN, H I 20 o HPLC-MS (Method C): m/z: 481 (M+1); Rt = 4.08 min.
WO 2004/080480 PCT/DK2004/000158 345 Example 890 (General procedure (N)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1 -yloxy]-N-hexylbutyramide o 0 HN , H 0 5 HPLC-MS (Method C): m/z: 441 (M+1); Rt = 4.31 min. Example 891 (General Procedure (N)) 4-({[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-7-carbonyl]amino}methyl)benzoic acid methyl ester O- CH 3 HN S H 0 N 0 HN 10 0 HPLC-MS (Method C): m/z: 436 (M+1); Rt.= 3.55 min. Example 892 (General procedure (N)) N-(4-Chlorobenzyl)-4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide H N N N, CI 0 15 HPLC-MS (Method C): m/z:493 (M+1); Rt = 4.19 min. Example 893 (General procedure (N)) N-(4-Chlorobenzyl)-3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide WO 2004/080480 PCT/DK2004/000158 346 HN N HN 0 CI HPLC-MS (Method C): m/z: 493 (M+1); Rt = 4.20 min. Example 894 (General Procedure (N)) 5 N-(4-Chlorobenzyl)-3-methyl-4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]benzamide HN NN N N O HK O' CH 3 NH
I
CI HPLC-MS (Method C): m/z: 507 (M+1); Rt = 4.37min. Example 895 (General procedure (N)) 10 5-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylamino)-isophthalic acid dimethyl ester
,CH
3 0 0 0 O /\ HN H 0 O .0
H
3 C HPLC-MS (Method C): m/z = 521 (M+1); Rt. = 4.57 min. 15 Example 896 (General procedure (N)) WO 2004/080480 PCT/DK2004/000158 347 5-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylamino}-isophthalic acid HO \0 0 HN H 0 O OH HPLC-MS (Method C): m/z = 515 (M+23); Rt. = 3.09 min. 5 Example 897 (General procedure (N)) 5-(3-{2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1 -yloxy]-ethyl}-ureido) isophthalic acid monomethyl ester - S N N oo
I-
NH 0 N'b 0 H 0 I HC' 10 HPLC-MS (Method C): m/z = 536 (M+1); Rt = 3,58 min. Example 898 (General Procedure (N)). 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester HN, N H
NCH
3 NN O / N3 0 15 4-[3-(1H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid (2.00 g, 5.41 mmol), 1 hydroxybenzotriazole (1.46 g, 10.8 mmol) and N,N-di(2-propyl)ethylamine (4.72 mL, 3.50 g, 27.1 mmol) were dissolved in dry N,N-dimethylformamide (60 mL). The mixture was cooled in an ice-water bath, and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (1.45 g, 7.56 mmol) and (S)-aminosuccinic acid dimethyl ester hydrochloride (1.28 g, 6.48 WO 2004/080480 PCT/DK2004/000158 348 mmol) were added. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 18 hours before it was poured into hydrochloric acid (0.1 N, 600 mL). The solid was collected by filtration and washed with water (2 X 25 mL) to furnish the title compound. 5 HPLC-MS (Method C): m/z: 513 (M+1); Rt = 3.65 min. 1 H-NMR (DMSO-d 6 ): 8 8.90 (1H, d), 8.86 (1H, d), 8.29 (1H, d), 8.11 (1H, dd), 7.87 (1H, d), 7.75 (2H, d), 7.69 (1H, d), 7.51 (1H, t), 7.32 (1H, t), 7.28 (2H, d), 5.82 (2H, s), 4.79 (1H, m), 3.61 (3H, s), 3.58 (3H, s), 2.92 (1H, dd), 2.78 (1H, dd). 10 Example 899 (General Procedure (N)). 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid HN. N H OH OH 0 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester (1.20 g, 2.34 mmol) was dissolved in tetrahydrofuran (30 mL). Aqueous sodium hydroxide (1 15 N, 14 mL) was added, and the resulting mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into hydrochloric acid (0.1 N, 500 mL). The solid was collected by filtration and washed with water (2 X 25 mL) and diethyl ether (2 X 25 mL) to furnish the title compound. HPLC-MS (Method C): m/z: 485 (M+1); Rt = 2.94 min. 20 'H-NMR (DMSO-d 6 ): 8 12.44 (2H, s (br)), 8.90 (1H, d), 8.68 (1H, d), 8.29 (1H, d), 8.11 (1H, dd), 7.87 (1H, d), 7.75 (2H, d), 7.68 (1H, d), 7.52 (1H, t), 7.32 (1H, t), 7.27 (2H, d), 5.82 (2H, s), 4.70 (1H, m), 2.81 (1H, dd), 2.65 (1H, dd). The compounds in the following examples were prepared in a similar fashion. 25 Example 900 (General procedure (N)) 2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-succinic acid dimethyl ester WO 2004/080480 PCT/DK2004/000158 349 HN NN NN HN 0 O ,, .CHz O HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3.65min. Example 901 (General procedure (N)) 5 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester HN, NN N ' N N ,CH -0 o Oo),CHz HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.57min. Example 902 (General procedure (N)) 10 (Methoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl)}-amino)-acetic acid methyl ester HN ,N N 0 ,CH3 N 0 HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3,55min. 15 Example 903 (General procedure (N)) 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid WO 2004/080480 PCT/DK2004/000158 350 N N . O OH O 0 0 OH HPLC-MS (Method C): m/z = 499 (M+1); Rt = 2.87min. Example 904 (General procedure (N)) 5 (Ethoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid ethyl ester N N OO O CHz O 0 1-CH, HPLC-MS (Method C): m/z = 541 (M+1); Rt = 3.91min. 10 Example 905 (General procedure (N)) 3-(3-{4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino} propylamino)-hexanedioic acid dimethyl ester HN H
H
3 H CH " o "CH HPLC-MS (Method C: m/z = 585 (M+1); Rt = 2,81 min. 15 Example 906 (General procedure (N)) 3-(3-{4-[4-(2,4-Dioxo-thiazol idin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino} propylamino)-hexanedioic acid WO 2004/080480 PCT/DK2004/000158 351 o O HO OH O 0 HPLC-MS (Method C): m/z = 554 (M-3); Rt = 3,19 min. Example 907 (General procedure (N)) 5 (Carboxymethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid HN.N N HPLC-MS (Method C): m/z = 485 (M+1); Rt = 3.04 min. Example 908 (General procedure (N)) 10 4-(3-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino} propylamino)-cyclohexane-1,3-dicarboxylic acid dimethyl ester HN * O HH O-
-
N .0 0 Hc.O 0 HPLC-MS (Method C): m/z = 612 (M+1); Rt = 3,24 min. 15 Example 909 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-hymethyl]benzoylamino}pentanedioic acid dimethyl ester 0O
HN
0o NL0 o z)CH H C 0 0 HPLC-MS (Method C): mlz = 6127 (M+1); Rt = 3,24 mi.
WO 2004/080480 PCT/DK2004/000158 352 Example 910 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester N,0 N O OCH, 0 HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min. 5 Example 911 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester Ns N 0 0 . O'CH 3 o 0 0 HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min. 10 Example 912 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid HN, N 0 OH No H yOH 0 HPLC-MS (Method C): m/z = 499 (M+1); Rt = 3.00 min. 15 Example 913 (General procedure (N)) (Methoxycarbonylmethyl-{3-[3-(2H-tetrazol-5-yl)carbazl-9-ylmethyl]benzoyl}amino)acetic acid methyl ester WO 2004/080480 PCT/DK2004/000158 353 HN' N N \/ \/N N 0 , 0 CH, N O'CH 3 0 0 1 H-NMR (DMSO-do): 5 8.88 (1H, d), 8.29 (1H, d), 8.10 (1H, dd), 7.85 (1H, d), 7.67 (1H, d), 7.52 (1H, t), 7.39 (1H, t), 7.30 (2H, m), 7.17 (2H, m), 5.79 (2H, s), 4.17 (2H, s), 4.02 (2H, s), 3.62 (3H, s), 3.49 (3H, s). 5 Example 914 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester N H OCH, N '0 s HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3.70 min. 10 Example 915 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-succinic acid HN N NN 0 OH HPLC-MS (Method C): m/z = 485 (M+1); Rt = 2.96 min. 15 Example 916 (General procedure (N)) (Carboxymethyl-{3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl}amino)acetic acid WO 2004/080480 PCT/DK2004/000158 354 N. HN N ./\ 0 OH o OH 0 HPLC-MS (Method C): m/z = 485 (M+1); Rt = 2.87 min. Example 917 (General procedure (N)) 5 4-(4-(3-Carboxy-propylcarbamoyl)4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl] benzoylamino}-butyrylamino)-butyric acid HN N NN N N OH OH The title compound was prepared by coupling of (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoylamino}pentanedioic acid bis-(2,5-dioxopyrrolidin-1-yl) ester (prepared from 10 (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid by essen tially the same procedure as described for the synthesis of 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoic acid 2,5-dioxopyrrolidin-1-yl ester) with 4-aminobutyric acid according to the procedure described for the preparation of 4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl] benzoylamino}butyric acid. 15 HPLC-MS (Method C): m/z: 669 (M+1); Rt = 2.84 min. Example 918 (General procedure (N)) [2-(2-{4-[3-(2H-Tetrazol-5-yI)-carbazol-9-ylmethyl]benzoylamino}ethoxy)ethoxy]acetic acid WO 2004/080480 PCT/DK2004/000158 355 -~ 0 NI, - -oH H>0 HN- O-,OH o HPLC-MS (Method C): mlz: 515 (M+1); Rt = 3.10 min. Example 919 (General procedure (N)) 5 2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-pentanedioic acid di-tert-butyl ester
N
HN N H 0 OH 3 N 0 O Hz CH3 HPLC-MS (Method C): m/z = 611 (M+1); Rt = 4.64 min. 10 Example 920 (General Procedure (N)). 4-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}butyric Acid ,Nz N HN N N OH N 0 HPLC-MS (Method C): m/z: 455 (M+1); Rt = 3.13 min. 15 Example 921 (General Procedure (N)). [2-(2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino)ethoxy)ethoxy]acetic acid WO 2004/080480 PCT/DK2004/000158 356 P~N ,N N HNN oOH 0 o The title compound was prepared by coupling of 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoic acid 2,5-dioxopyrrolidin-1-yl ester with [2-(2-aminoethoxy)ethoxy]acetic acid (prepared from [2-[2-(Fmoc-amino)ethoxy]ethoxy]acetic acid by treatment with PS-Trisamine 5 resin in DMF). HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.10 min. The commercially available compounds in the following examples do all bind to the HisB10 Zn 2 +site: 10 Example 922 1-(4-Bromo-3-methylphenyl)-1,4-dihydrotetrazole-5-thione
,N
N HN ' Y N SN Br
CH
3 15 Example 923 1-(4-lodophenyl)-1,4-dihydrotetrazole-5-thione N=N HN S Example 924 20 1-(2,4,5-Trichlorophenyl)-I H-tetrazole-5-thiol
,N'~
N HN N C HN Cl SC Cl CI CI WO 2004/080480 PCT/DK2004/000158 357 Example 925 1-(2,6-Dimethylphenyl)-1,4-dihydrotetrazole-5-thione NN CH 3 HN N Y)IN S H3C 5 Example 926 I -(2,4,6-Trimethylphenyl)-1,4-dihydrotetrazole-5-thione ,NN CH 3 HN
H
3 C CH 3 Example 927 10 1-(4-Dimethylaminophenyl)-1lH-tetrazole-5-thiol ,NN
HN
S
N.CH
3
CH
3 Example 928 1-(3,4-Dichlorophenyl)-1,4-dihydro-1H-tetrazole-5-thione ,NzN HN ' SCI I cl 15 cl Example 929 1-(4-Propylphenyl)-1,4-dihydro-1H-tetrazole-5-thione N N HN 'j I SN
CH
3 20 WO 2004/080480 PCT/DK2004/000158 358 Example 930 1-(3-Chlorophenyl)-1,4-dihydro-1 H-tetrazole-5-thione ,NzN HN SN I CI 5 Example 931 1-(2-Fluorophenyl)-1,4-dihydro-1 H-tetrazole-5-thione ,N-N HN I F Example 932 10 1-(2,4-Dichlorophenyl)-1,4-dihydro-1lH-tetrazole-5-thione ,N N HN I S N Sc ci Example 933 1-(4-Trifluoromethoxyphenyl)-1,4-dihydro-1 H-tetrazole-5-thione ,N N HN SrN OF 15 F Example 934 N-[4-(5-Mercaptotetrazol-1-yl)-phenyl]-acetamide N N HN N SN NH
H
3 C 0 20 WO 2004/080480 PCT/DK2004/000158 359 Example 935 1-(4-Chlorophenyl)-1,4-dihydrotetrazole-5-thione HN ' HN SN CI 5 Example 936 1-(4-Methoxyphenyl)-1,4-dihydrotetrazole-5-thione N N HN S O,CH3 Example 937 10 1-(3-Fluoro-4-pyrrolidin-1 -ylphenyl)-1,4-dihydrotetrazole-5-thione N N HN Example 938 N-[3-(5-Mercaptotetrazol-1 -yl)phenyl]acetamide N-N N"N SH do N ' CH3 15 H Example 939 1-(4-Hydroxyphenyl)-5-mercaptotetrazole WO 2004/080480 PCT/DK2004/000158 360 /N N N\ HS OH Example 940 NzN 0 N N 0
CH
3 SH 5 Preparation of 1-aryl-1,4-dihydrotetrazole-5-thiones (or the tautomeric 1-aryltetrazole-5 thiols) is described in the literature (eg. by Kauer & Sheppard, J. Org. Chem., 32, 3580-92 10 (1967)) and is generally performed eg. by reaction of aryl-isothiocyanates with sodium azide followed by acidification 1-Aryl-1,4-dihydrotetrazole-5-thiones with a carboxylic acid tethered to the aryl group may be prepared as shown in the following scheme: 15 9+ 0+ oN Step1 N (CH 2 * Step 2 H2NOH OHO OH 0 o " Y oH.OH 0 0 N N1Step 3 ,N=N Ni N. (CStep4 SON SNH2 OH S I CN (CH 2
)
m O O O Step 1 is a phenol alkylation and is very similar to steps 1 and 2 of general procedure (D) and may also be prepared similarly as described in example 481.
WO 2004/080480 PCT/DK2004/000158 361 Step 2 is a reduction of the nitro group. SnCl 2 , H 2 over Pd/C and many other procedures known to those skilled in the art may be utilised. Step 3 is formation of an arylisothiocyanate from the corresponding aniline. As reagents CS 2 , 5 CSCl2, or other reagents known to those skilled in the art, may be utilised. Step 4 is a conversion to mercaptotetrazole as described above. Compounds of the invention include: Example 941 Example 942 ,N N HN 'N HN IN S NS Example 943 Example 944 ,N:N NzN HN HN N O OH S O OH O O Example 945 Example 946 ,N::N *NzN HN ' HNO N 0 OH OH S~ OH 0 Example 947 ,NzN HN S OH O 10 Example 948 4-(4-Hydroxyphenyl)-1 H-[1,2,3]triazole-5-carbonitrile WO 2004/080480 PCT/DK2004/000158 362 HO N
-
\_NH N Phenylsulphonyl acetonitrile (2.0 g, 11.04 mmol) was mixed with 4-hydroxybenzaldehyde (1.35 g, 11.04 mmol) in DMF (10 mL) and toluene (20 mL). The mixture was refluxed for 3 hours and subsequently evaporated to dryness in vacuo. The residue was treated with di 5 ethyl ether and toluene. The solid formed was filtered to afford 2.08 g (66%) of 2 benzenesulfonyl-3-(4-hydroxyphenyl)acrylonitrile. HPLC-MS (Method C): m/z: 286 (M+1); Rt. = 3.56 min. A mixture of 2-benzenesulfonyl-3-(4-hydroxyphenyl)acrylonitrile (2.08 g, 7.3 mmol) and so 10 dium azide (0.47g,7.3 mmol) in DMF (50 mL) was heated at reflux temperature 2 hours. After cooling, the mixture was poured on ice. The mixture was evaporated in vacuo to almost dry ness and toluene was added. After filtration, the organic phase was evaporated in vacuo. The residue was purified by silicagel chromatography eluting with a mixture of ethyl acetate and heptane (1:2). This afforded 1.2 g (76%) of the title compound. 15 1H NMR (DMSO-d): 10.2 (broad,1H); 7.74 (d,2H); 6.99 (d,2H); 3.6-3.2 (broad,1H). HPLC-MS (Method C) m/z: = 187 (M+1); Rt. = 1.93 min General procedure (0) for preparation of compounds of general formula 114: NN O N AN N oO o o , N=N O=S H = HN + o= AA AA II Step 1 Step 2 N 20 114 wherein AA is as defined above, 25 Steps 1 and 2 are described in the literature (eg Beck & GOnther, Chem. Ber., 106, 2758-66 (1973)) WO 2004/080480 PCT/DK2004/000158 363 Step 1 is a Knoevenagel condensation of the aldehyde AA-CHO with phenylsulfonyl acetonitrile and step 2 is a reaction of the vinylsulfonyl compound obtained in step 1 with so dium azide. This reaction is usually performed in DMF at 90 - 110 OC. 5 This general procedure is further illustrated in the following example 949: Example 949 (General Procedure (0)) [4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)phenoxy]acetic acid HNN- N 0 O 10 HO Phenylsulphonylacetonitrile (0.1 g, 0.55 mmol) was mixed with 4-formylphenoxyactic acid (0.099 g, 0.55 mmol) in DMF (3 mL) and heated to 110 oC for 3 h and subsequently cooled to RT. Sodium azide (0.036 g, 0.55 mmol) was added and the resulting mixture was heated to 110 'C for 3 h and cooled to RT. The mixture was poured into water (20 mL) and centrifuged. 15 The supernatant was discarded, ethanol (5 mL) was added and the mixture was centrifuged again. After discarding the supernatant, the residue was dried in vacuo to afford 50 mg (37%) of [4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)phenoxy]acetic acid. HPLC-MS (Method C): m/z: 245 (M+1) Rt. 2.19 min. 20 Example 950 (General Procedure (0)) 5-(Naphthalen-1 -yl)-3H-[1,2,3]triazole-4-carbonitrile HNN- N NZ HPLC-MS (Method C): m/z: 221 (M+1); Rt. 3.43 min. 25 WO 2004/080480 PCT/DK2004/000158 364 Example 951 (General Procedure (0)) 5-(Naphthalen-2-yl)-3H-[1,2,3]triazole-4-carbonitrile HN N NZ HPLC-MS (Method C): m/z: 221 (M+1I); Rt = 3.66 min. 5 Example 952 (General procedure (0)) 4-[3-(5-Cyano-[1,2,3]triazol-4-yl)-1,4-dimethylcarbazol-9-ylmethyl]-benzoic acid HN NN HO CH 3/\C0 H 3 C
-
7 N HPLC-MS (Method C): m/z = 422 (M+1); Rt = 3.85 min. '10 Preparation of intermediary aldehyde: 1,4 Dimethylcarbazol-3-carbaldehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mrnix ture was stirred for 1 hour at room temperature. 4-Bromomethylbenzoic acid (0.73 g, 3.4 15 mmol) was slowly added and the resulting slurry was heated to 40 °C for 16 hours. Water (5 mL) and hydrochloric acid (6N, 3 mL) were added. After stirring for 20 min at room tempera ture, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formyl-1,4-dimethylcarbazol-9-ylmethyl)benzoic acid. 20 HPLC-MS (Method C): m/z = 358 (M+1), RT. = 4.15 min. Example 953 (General Procedure (0)) 5-(Anthracen-9-yl)-3H-[1,2,3]triazole-4-carbonitrile WO 2004/080480 PCT/DK2004/000158 365 HNN N N HPLC-MS (Method C): m/z: 271 (M+1); Rt = 3.87 min. Example 954 (General Procedure (0)) 5 5-(4-Methoxynaphthalen-1 -yl)-3H-[1,2,3]triazole-4-carbonitrile HN N
N
/ 0
H
3 C HPLC-MS (Method C): m/z: 251 (M+1); Rt = 3.57 min. Example 955 (General Procedure (0)) 10 5-(1,4-Dimethyl-9H-carbazol-3-yl)-3H-[1,2,3]triazole-4-carbonitrile 'N* HNN N NZ HC CH 3 H3 3 NH HPLC-MS (Method C): m/z: 288 (M+1); Rt = 3.67 min. Example 956 (General procedure (0)) 15 5-(4'-Methoxybiphenyl-4-yl)-3H-[1,2,3]triazole-4-carbonitrile WO 2004/080480 PCT/DK2004/000158 366 HN ,N"N NO 0 HaC HPLC-MS (Method C): m/z = 277 (M+1); Rt = 3.60 min. Example 957 (General procedure (0)) 5 5-(4-Styrylphenyl)-3H-[1,2,3]triazole-4-carbonitrile
N
HNN N HPLC-MS (Method C): m/z = 273 (M+1); Rt = 4.12 min. Example 958 (General procedure (0)) 10 5-(2,6-Dichloro-4-dibenzylaminophenyl)-3H-[1,2,3]triazole-4-carbonitrile HN-f N N Cl CI NHPLC-MS (Method C): mz = 434 (M); Rt 4.64 in. HPLC-MS (Method C): m/z = 434 (M+1); Rt = 4.64 min.
WO 2004/080480 PCT/DK2004/000158 367 Example 959 (General procedure (0)) 5-(1 -Bromonaphthalen-2-yl)-3H-[1,2,3]triazole-4-carbonitrile Br K N- N S/ N H N 5 HPLC-MS (Method C: m/z = 300 (M+1); Rt. = 3.79 min. Example 960 4-(4-Bromophenyl)-1 H-[1,2,3]triazole-5-carbonitrile -* Br N H N 10 This compound is commercially available (MENAI). Example 961 N-[4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)-phenyl]-acetamide H N N / -CH 3 N1/ NOH N H N 15 This compound is commercially available (MENAI). Example 962 (General procedure (0)) WO 2004/080480 PCT/DK2004/000158 368 5-(4'-Chlorobiphenyl-4-yl)-3H-[1,2,3]triazole-4-carbonitrile HN N N N_ Cl HPLC-MS (Method C): m/z = 281 (M+1); Rt = 4.22 min. 5 The compounds in the following examples are commercially available and may be prepared using a similar methodology: Example 963 4-(4-Trifluoromethoxyphenyl)-1 H-[1,2,3]triazole-5-carbonitrile F F F 0 Nq N N 10 H Example 964 4-Benzo[1,3]dioxol-5-yl-1H-[1,2,3]triazole-5-carbonitrile N oN 15 Example 965 4-(3-Trifluoromethylphenyl)-1 H-[1,2,3]triazole-5-carbonitrile WO 2004/080480 PCT/DK2004/000158 369 F F F -N N N---NH Example 966 4-Pyridin-3-yl-1 H-[1,2,3]triazole-5-carbonitrile NH N N 5 N Example 967 4-(2,6-Dichlorophenyl)-1 H-[1,2,3]triazole-5-carbonitrile Cl cii NC % NH \NN 10 Example 968 4-Thiophen-2-yl-1H-[1,2,3]triazole-5-carbonitrile K S N N a e N N H 15 Example 969 WO 2004/080480 PCT/DK2004/000158 370 3,5-Dimethylisoxazole-4-carboxylic acid 4-(5-cyano-1H-[1,2,3]triazol-4-yl)phenyl ester CH 3
CH
3 0 NH N=N Example 970 5 3,3-Dimethyl-butyric acid 4-(5-cyano-1H-[1,2,3]triazol-4-yl)phenyl ester CH3 HC CH, N 0 NH N N Example 971 4-Methyl-[1,2,3]thiadiazole-5-carboxylic acid 4-(5-cyano-1 H-[1,2,3]triazol-4-yl)phenyl ester H N N N 0 10
CH
3 Example 972 4-Chlorobenzoic acid 4-(5-cyano-1 H-[1,2,3]triazol-4-yl)phenyl ester H N N
CI
WO 2004/080480 PCT/DK2004/000158 371 Example 973 4-(3-Phenoxyphenyl)- 1H-[1,2,3]triazole-5-carbonitrile NN\ I ~NH ON 5 Example 974 4-(5-Bromo-2-methoxyphenyl)-1 H-[1,2,3]triazole-5-carbonitrile ,CH 3 NH N Br 10 Example 975 4-(2-Chloro-6-fluorophenyl)-1 H-[1,2,3]triazole-5-carbonitrile F N:N 'NH ~N CI The following cyanotriazoles are also compounds of the invention: 15 4-(2-Chloro-6-fluorophenyl)-1 H-[1,2,3]triazole-5-carbonitrile. Terephthalic acid mono[ 4-(5-cyano-1lH-[1,2,3]triazol-4-yl)phenyl] ester. N- [4-(5-cyano-1lH-[1,2,3]triazol-4-yl)-phenyl]terephthalamic acid 4-(4-Octyloxyphenyl)-1 H-[1,2,3]triazole-5-carbonitrile 20 4-(4-Styrylphenyl)-1H-[1,2,3]triazole-5-carbonitrile. 4-(4'-Trifluoromethylbiphenyl-4-yl)-1 H-[1,2,3]triazole-5-carbonitrile. 4-(4'-Chlorobiphenyl-4-yl)-1 H-[1,2,3]triazole-5-carbonitrile. 4-(4'-Methoxybiphenyl-4-yl)-1 H-[1,2,3]triazole-5-carbonitrile. 4-(1-Naphthyl)-l H-[1,2,3]triazole-5-carbonitrile.
WO 2004/080480 PCT/DK2004/000158 372 4-(9-Anthranyl)-1 H-[1,2,3]triazole-5-carbonitrile. 4-(4-Methoxy-1 -naphthyl)-1 H-[1,2,3]triazole-5-carbonitrile. 4-(4-Aminophenyl)- 1 H-[1,2,3]triazole-5-carbonitrile. 4-(2-Naphthyl)- 1 H-[1,2,3]triazole-5-carbonitrile. 5 General procedure (P) for preparation of compounds of general formula 115: O /OH + L O Step 1 O O 'R" Step 2 O OH +Lea \ 0 R" O1 0 H 0 H 0 H 0 I Step 3
SO
2 Ph S02Ph
(CH
2 ) NN (CH 2 ). Step 4 A / 2) OH HN o T\OH d 0 lo N 115 10 wherein n is 1 or 3-20, AA is as defined above, R" is a standard carboxylic acid protecting group, such as Cl-C 6 -alkyl or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the 15 like. This procedure is very similar to general procedure (D), steps 1 and 2 are identical. Steps 3 and 4 are described in the literature (eg Beck & G0nther, Chem. Ber., 106, 275a-66 20 (1973)) Step 3 is a Knoevenagel condensation of the aldehyde obtained in step 2 with phenylsulfon ylacetonitrile and step 4 is a reaction of the vinylsulfonyl compound obtained in step 3 with sodium azide. This reaction is usually performed in DMF at 90 - 110 OC.
WO 2004/080480 PCT/DK2004/000158 373 This General procedure (P) is further illustrated in the following two examples Example 976 (General procedure (P)) 5 5-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentanoic acid ethyl ester O -CH 3 0 0 N H N 6-Hydroxynaphthalene-2-carbaldehyde (Syncom BV. NL, 15.5 g, 90 mmol) and K 2
CO
3 (62.2 g, 450 mmol) were mixed in DMF (300mL) and stirred at room temperature for 1hour. Ethyl 5-bromovalerate (21.65 g, 103.5 mmol) was added and the mixture was stirred at room tern 10 perature for 16 hours. Activated carbon was added and the mixture was filtered. The filtrate was evaporated to dryness in vacuo to afford 28.4 g of crude 5-(6-formylnaphthalen-2 yloxy)pentanoic acid ethyl ester, which was used without further purification. HPLC-MS (Method C ): m/z = 301 (M+1); Rt. = 4.39 min. 15 5-(6-Formylnaphthalen-2-yloxy)pentanoic acid ethyl ester (28.4 g, 94.5 mmol), phenylsulfon ylacetonitrile (20.6 g, 113.5 mmol), and piperidine (0.94 mL) were dissolved in DMF (200 mL) and the mixture was heated at 50 °C for 16 hours. The resulting mixture was evaporated to dryness in vacuo and the residue was dried for 16 hours at 40 0C in vacuo. The solid was 20 recrystallised from 2-propanol (800 mL) and dried again as described above. This afforded 35 g (80%) of 5-[6-(2-benzenesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester. HPLC-MS (Method C): m/z = 486 (M+23); Rt. = 5.09 min. 25 5-[6-(2-Benzenesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester (35 g, 74.6 mmol) and sodium azide (4.9 g, 75.6 mmol) were dissolved in DMF (100 mL) and stirred WO 2004/080480 PCT/DK2004/000158 374 for 16 hours at 50 'C. The mixture was evaporated to dryness in vacuo, redissolved in THF / ethanol and a small amount of precipitate was filtered off. The resulting filtrate was poured into water (2.5 L). Filtration afforded after drying 24.5 g (88%) of 5-[6-(5-cyano-1H [1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoic acid ethyl ester (24.5 g, 88%). 5 HPLC-MS (Method C): m/z = 365 (M+1); Rt. = 4.36 min. Example 977 (General procedure (B)) 5-[6-(5-Cyano-1 H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentanoic acid HNN 100 10 N o _'OH 5-[6-(5-Cyano-1lH-[1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoicacid ethyl ester (24.5 g, 67.4 mmol) was dissolved in THF (150 mL) and mixed with sodium hydroxide (8.1 g, 202 mmol) dissolved in water (50 mL). The mixture was stirred for 2 days and the volatiles were evaporated in vacuo. The resulting aqueous solution was poured into a mixture of water (1 L) 15 and hydrochloric acid (1N, 250 mL). The solid was isolated by filtration, dissolved in sodium hydroxide (1N, 200 mL), and the solution was washed with DCM and then ethyl acetate, the aquous layer was acidified with hydrochloric acid (12N). The precipitate was isolated by filtra tion, dissolved in THF / diethyl ether, the solution was treated with MgSO 4 and activated car bon, filtrated and evaporated in vacuo to almost dryness followed by precipitation by addition 20 of pentane (1 L). This afforded after drying in vacuo 17.2 g (76%) of the title compound. HPLC-MS (Method C): m/z = 337 (M+1); Rt. = 3.49 min. Example 978 (General procedure (P)) 25 6-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)naphthalen-2-yloxy]hexanoic acid WO 2004/080480 PCT/DK2004/000158 375 0 OH 0 N N' N H ~ HPLC-MS (Method C): m/z = 351 (M+1I); Rt = 3.68 min. Example 979 (General procedure (P)) 5 11-[6-(5-Cyano- 1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-undecanoic acid
N
N HN' " 0 N H O-- OH HPLC-MS (Method C): m/z = 443 (M+23); Rt = 4.92 min. Example 980 (General procedure (P)) 10 2-{3-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-propyl}-malonic acid diethyl ester N -N HzC 0 CH, HPLC-MS (Method C): m/z = 465 (M+1); Rt. = 4.95 min. Example 981 (General procedure (P)) 15 2-{5-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentyl}-malonic acid diethyl ester WO 2004/080480 PCT/DK2004/000158 376 HN N N 0
O
ooJ 0 0 0 HPLC-MS (Method C): m/z = 465 (M+1); Rt. = 4.95 min. Example 982 (General procedure (P)) 5 2-{3-[6-(5-Cyano-1H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-propyl}-malonic acid N::N O O O N 0 0 HPLC-MS (Method C): m/z = 381 (M+1); Rt. = 3.12 min. Example 983 (General procedure (P)) 10 2-{5-[6-(5-Cyano-1 H-[1,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentyl}-malonic acid NN NO NJN 0 - 0 0 0 0 HPLC-MS (Method C): m/z 0 409 (M+1); Rt. = 3.51 min. Example 984 (General procedure (P)) 15 4-[4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)-phenoxy]butyric acid WO 2004/080480 PCT/DK2004/000158 377 HN-N N O 0 HO 0 HPLC-MS (Method C): m/z = 273 (M+1); Rt = 2.44 min. The following compounds may be prepared according to this general procedure (P): 5 4-(4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)phenoxy)butyric acid: N=N HN - I Io N-N HN I 10 0 I0 / O OH OH N= 0 4-(4-(5-Cyano-1H--[1,2,3]triazol-4-yl)phenoxy)butyric acid ethyl ester 5-(4-(5-Cyano-1H-[1,2,3]triazol-4-yl)phenoxy)pentanoic acid 8-(4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)phenoxy)octanic acid IN=N HN , 10 0 1-(4-(5-Cyano-1 H-[1,2,3]triazol-4-yl)phenoxy)utyrdecanic acid ethyl ester 15 12-(4-(5-Cyano-1H-[1,2,3]triazol-4-yl)phenoxy)dodecnanoic acid 8-(4-(5-Cyano- I H-[i ,2,3]triazol-4-yI)phenoxy)octanoic acid 1 0-(4-(5-Cyano-1 H-[I ,2,3]triazol-4-yI)phenoxy)decanoic acid 15 1 2-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yI)phenoxy)dodecanoic acid WO 2004/080480 PCT/DK2004/000158 378 General procedure (R) for preparation of compounds of general formula 112: ,N N R 3 ,N N R 3 HNI. . R Pol-Cl /DIEA POI-N. - i:DMAP/DIPEA / DMF IN 2 R N 2-_ _ DMF /DCM R N N ii: HT-COCI H H ,NN R 3 NzN R 3 Pol-N, . HN. / -N 2 .1 N HC in N R 2 RaR 2 N THF / Et 2 O I EtOH = 8: :1 N PT-H T-H O O 112 I PoI- = PS -0 C cI 5 wherein T is as defined above and R 2 and R 3 are hydrogen, aryl or lower alkyl, both option ally substituted. The general procedure (R) is further illustrated by the following example: 10 Example 985 (General procedure (R)) Phenyl-[3-(2H-tetrazol-5-yl)-carbazol-9-yl]-methanone N
-
N HNNI 2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloro methane (4 mL) for 30 minutes. The solvent was drained, and a solution of 3-(2H-tetrazol-5 15 yl)-9H-carbazole (80 mg, 0.34 mmol) in a mixture of N,N-dimethylformamide / dichloro methane / N,N-di(2-propyl)ethylamine (5:5:1) (3 mL) was added. The reaction mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed thoroughly with N,N-dimethylformamide (2 x 4 mL) and dichloromethane WO 2004/080480 PCT/DK2004/000158 379 (6 x 4 mL). A solution of 4-(dimethylamino)pyridine (14 mg, 0.11 mmol) and N,N-di(2 propyl)ethylamine (0.23 mL, 171 mg, 1.32 mmol) in N,N-dimethylformamide (2 mL) was added followed by benzoyl chloride (0.13 mL, 157 mg, 1.12 mmol). The mixture was shaken for 48 hours at room temperature. The drained resin was washed consecutively with di 5 chloromethane (2 x 4 mL), methanol (2 x 4 mL) and tetrahydrofuran (4 mL). The resin was treated for 2 hours at room temperature with a solution of dry hydrogen chloride in tetrahy drofuran / ethyl ether / ethanol = 8:1:1 (0.1 M, 3 mL). The reaction mixture was drained and concentrated. The crude product was stripped with dichloromethane (1.5 mL) three times to yield the title compound. 10 HPLC-MS (Method C): m/z: 340 (M+1); Rt = 3.68 min. 1 H-NMR (DMSO-d6): 8 8.91 (1H, s), 8.34 (1H, d), 8.05 (1H, d), 7.78 (3H, m), 7.63 (3H, m), 7.46 (2H, m), 7.33 (1H, dd). 15 The compounds in the following examples were prepared in a similar fashion. Example 986 (General procedure (R)) Phenyl-[5-(2H-tetrazol-5-y)-indol-1 -yl]-methanone , N -'N HN, N N N 0 20 HPLC-MS (Method C): m/z: 290 (M+1); Rt = 3.04 min. 1 H-NMR (DMSO-d 6 ): 8 8.46 (1H, d), 8.42 (1H, d), 8.08 (1H, dd), 7.82 (2H, d), 7.74 (1H, t), 7.64 (2H, t), 7.55 (1H, d), 6.93 (1H, d). Example 987 (General procedure (R)) 25 (2,3-Difluorophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone WO 2004/080480 PCT/DK2004/000158 380 HN HNNF N o F HPLC-MS (Method B): m/z = 326 (M+1); Rt = 3.85 min. Example 988 (General procedure (R)) 5 (2-Fluoro-3-trifluoromethylphenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone fN-- N HN N F F F 0F HPLC-MS (Method B): m/z = 376 (M+1); Rt = 4.32 min. Example 989 (General procedure (R)) 10 (3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone HN,N :. N HNN O . O o4:Z 0 HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min. Example 990 (General procedure (R)) 15 (4-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone HN ,N .:J NO Ho3 oo 0 HPLC-MS (Method B3): mlz = 335 (M+1); Rt = 3.71 min.
WO 2004/080480 PCT/DK2004/000158 381 Example 991 (General procedure (R)) Naphthalen-2-yl-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone HN N . HN, N N H, 5 HPLC-MS (Method C): m/z: = 354 (M+1); Rt = 3.9125 min. 10 Example 993 (General procedure (R)) HNN N /OH 3 Br HPLC-MS (Method C): m/z: 3418 (M+1); Rt = 4.39 min. 10 15 Example 994 (General procedure (R)) / Nz--N HN N 9 N -" N o O-CH3 HPLC-MS (Method C): m/z: 370 (M+1); Rt = 4.01 min.
WO 2004/080480 PCT/DK2004/000158 382 Example 995 (General procedure (R)) H N N / NO N 0 CI HPLC-MS (Method C): m/z: 374 (M+1); Rt = 4.28 min. 5 Example 996 (General procedure (R)) NZN HN N N 0 HPLC-MS (Method C): m/z: 416 (M+1); Rt = 4.55 min. 10 Example 997 (General procedure (R)) 7 N HN . N 0
H
3 C HPLC-MS (Method C): m/z: 354 (M+1); Rt = 4.22 min.
WO 2004/080480 PCT/DK2004/000158 383 Example 998 (General procedure (R)) HN-N. N \/O HPLC-MS (Method C): m/z: 358 (M+1); Rt = 3.91 min. 5 Example 999 (General procedure (R)) HN NO 0 HPLC-MS (Method C): m/z: 390 (M+1); Rt = 4.38 min. Example 1000 (General procedure (R)) HN NO N 0 10 Br HPLC-MS (Method C): m/z: 418 (M+1); Rt = 4.36 min. Example 1001 (General procedure (R)) HNN N <N N 3 Se
CH
3 15 HPLC-MS (Method C): mfz: 304 (M+1); Rt = 3.32 min.
WO 2004/080480 PCT/DK2004/000158 384 Example 1002 (General procedure (R)) ,N- N HN N N N 0 Br HPLC-MS (Method C): mlz: 368 (M+1); Rt = 3.84 min. 5 Example 1003 (General procedure (R)) N N HN N "N O 0
H
3 C HPLC-MS (Method C): m/z: 320 (M+1); Rt = 3.44 min. Example 1004 (General procedure (R)) ,N-N HN, N SN 0 10 CI HPLC-MS (Method C): m/z: 324 (M+1); Rt = 3.73 min.
WO 2004/080480 PCT/DK2004/000158 385 Example 1005 (General procedure (R)) HN :: N'N HN NN o /CH 3 HPLC-MS (Method C): mlz: 304 (M+1); Rt = 3.64 min. 5 Example 1006 (General procedure (R)) HN, NN HN'N N O HPLC-MS (Method A): m/z: 308 (M+1); Rt = 3.61 min. Example 1007 (General procedure (R)) HN N N 0 10 Br HPLC-MS (Method C): m/z: 368 (M+1); Rt = 3.77 min. Example 1008 (General procedure (R)) ,NN HNN 15 HPLC-MS (Method A): (sciex) m/z: 326 (M+1); Rt = 3.73 min. 15 HPLC-MS (Method A): (sciex) m/z: 326 (M I); Rt = 3.73 mai.
WO 2004/080480 PCT/DK2004/000158 386 HPLC-MS (Method C): m/z: 326 (M+1); Rt = 3.37 min. Example 1009 (General procedure (R)) /N-N HN,N N NN 0 F F F 5 HPLC-MS (Method C): m/z: 374 (M+1); Rt = 4.03 min. General procedure (Q) for preparation of compounds of general formula 116s: H2N-PS
H
2 N-(Arg)-N-PS H
H
2 N-(Gly)m- (Arg); N-PS
H
2 N -(Abz);-(Gly) (Arg)N - PS N N D-(4Abz)/Gly)gArg);N - P S H N I H D N NH-(4-Abz)p(Gly)gArg);NH 2
N
I 10 H 116 wherein PS is polymeric support, a Tentagel S RAM resin, n is 1 - 20, m is 0 - 5, and p is 0 or 1.
WO 2004/080480 PCT/DK2004/000158 387 The compounds of the invention of general formula (12) can be prepared by means of stan dard peptide chemistry (General procedure H), e.g. in 0.5 mmol scale, using Fmoc strategy and HOAt or HOBT activated amino acids. The compounds prepared in the following exam ples according to General procedure (Q) were all isolated as the TFA salts. This procedure is 5 further illustrated in the following: Typically, 2 gram of Fmoc Tentagel S RAM resin (Rapp Polymere, Tubingen) with substitu tion 0,25 mmol/g was washed with NMP then treated with 25% piperidine in NMP for 30 rnin followed by wash with NMP which renders the resin ready for coupling. Step wise coupling of Fmoc-Arginine (Fmoc-Arg(Pmc)-OH), Fmoc-Glycine (Fmoc-Gly-OH) 10 and Fmoc-4-aminobenzoic acid (Fmoc-4-Abz-OH): To 2 mmol of Fmoc-L-Arg(Pmc)-OH (Novabiochem) was added 3,33 ml 0,6M HOAt in NMP (Perseptives) or 0,6M HOBT in NMP (Novabiochem) containing 0,2% bromphenolblue as indicator and added 330 il of diisopropylcarbodiimide DIC (Fluka) and the solution was then added to the resin. After coupling for minimum 1 hour, or when the blue colour disappeared, 15 the resin was washed with NMP and the Fmoc group was deprotected with 25% piperidine in NMP for 20 minutes followed by wash with NMP. This stepwise assembling of the arginine residues was repeated to give 3, 4, 5 or 6 arginines on the resin. The Fmoc-Glycine (No vabiochem) and Fmoc-4-aminobenzoic acid (Fluka and Neosystems) were coupled using the same procedure as described for Fmoc-Arg(Pmc)-OH. 20 Coupling of A-OH, e.g. 1H-benzotriazole-5-carboxylic acid on Gly. When A-OH, e.g. 1H-benzotriazole-5-carboxylic acid (Aldrich) was coupled on a glycine or arginine residue the coupling procedure was as described above. Coupling of A-OH, e.g. 1H-benzotriazole-5-carboxylic acid on Abz or 4-Apac: Due to the lower nucleophilicity of the amino group in Abz the following procedure was nec 25 essary. To 4 mmol of A-OH, e.g. 1H-benzotriazole-5-carboxylic acid was added 6,66 ml of a solution of 0,6M HOAt, 0,2 mmol dimethylaminopyridine (DMAP) and 4 mmol DIC and was then added to the resin and allowed to react overnight. Introduction of fragment 4-Apac instead of 4-Abz: 30 4-Nitrophenoxyacetic acid may be coupled on a glycine or arginine residue using DIC and HOBT/HOAt as described above. Subsequent reduction of the nitro group may be done IJs ing SnCl 2 in NMP or DMF e.g. as described by Tumelty et al. (Tet. Lett., (1998) 7467-70). Cleavage of the peptides from the resin. After synthesis the resin was washed extensively with diethyl ether and dried. To 1 grarr of 35 the peptidyl resin was added 25 ml of a TFA solution containing 5% thioanisole, 5% ethanol, WO 2004/080480 PCT/DK2004/000158 388 5% phenol and 2% triisopropylsilane and allowed to react for 2 hours. The TFA solution was filtered and concentrated with argon flow for approximately 30 minutes. Then diethylether ca. 5-7 times the residual volume of TFA was added and the peptide precipitate was extracted in 10% AcOH and washed 5 times with diethyl ether and lyophilized. 5 RP-HPLC analysis and purification: The crude products were analysed on RP-HPLC C18 column (4,6 x 250 mm) using one of two gradients (see table 1 and 2), temperature 25 0 C, wavelength 214 nm and flow rate 1 ml/min with A-buffer 0,15 % (W/w) TFA in H 2 0 and B Buffer (87,5 % (W/w) MeCN, 0,13 % (W/w) TFA in H 2 0). 10 The products were purified on preparative RP-HPLC C18 column (2x25 cm) using a gradient (variable, see e.g example 1013 and similar), temperature 25 0 C, wavelength 214 nm and flow rate 6 mI/min with A-buffer 0,15 % (W/w) TFA in H 2 0 and B-Buffer (87,5 % (W/w) MeCN, 0,13 % (W/w) TFA in H 2 0) and verified by mass spectrometry (MALDI). 15 Table 1: Time (min.) Flow (ml/min) %A %B ( 0 1,00 95,0 5,0 30,00 1,00 80,0 20,0 35,00 1,00 0,0 100,0 40,00 1,00 0,0 100,0 45,00 1,00 95,0 5,0 Table 2: Time (min.) Flow (mI/min) %A %B 0 1,00 95,0 5,0 30,00 1,00 40,0 60,0 31,00 1,00 0,00 100,0 35,00 1,00 0,00 100,0 WO 2004/080480 PCT/DK2004/000158 389 36,00 1,00 95,0 5,0 The following examples were prepared using this general procedure (0). Example 1010 (General Procedure (Q)) 5 Benzotriazol-5-ylcarbonyl-Gly 2 -Arg 3
-NH
2
(BT-G
2
R
3 ). HN NH 2 HNy NH 2 NH NH N N- N NH2 0 H 0 NH H HN H 2 N HN HNNH MS (MALDI): m/z: 746.7 g/mol; calculated: 744.2 g/mol. HPLC gradient: Time (min) Flow %A %B (ml/min) 0,00 6,00 90,0 10,0 120,00 6,00 90,0 10,0 121,00 0,10 90,0 10,0 10 Example 1011 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-Gly 2 -Arg 4
-NH
2
(BT-G
2 R4)-. HN yNH 2 HN yNH 2 NH NH N-N N N NH 2 'N 0 0 HN HN
H
2 N NH H 2 N 'NH MS (MALDI): m/z: 903.0 g/mol; calculated: 900.6 g/mol. 15 HPLC gradient: WO 2004/080480 PCT/DK2004/000158 390 Time (min) Flow %A %B (ml/min) 0,00 6,00 95,0 5,0 30,00 6,00 80,0 20,0 35,00 6,00 0,0 100,0 40,00 6,00 0,0 100,0 45,00 6,00 95,0 5,0 64,00 6,00 95,0 5,0 Example 1012 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-Gly 2 -Args-NH 2
(BT-G
2
R
5 ). HN NH 2 HN NH 2 HN NH 2 NH NH NH O OH O N N N N N N O N H N H 2 N H H - H 0 - H HN HN 5
H
2 N NH H 2 N NH MS (MALDI): m/z: 1060.8 g/mol; calculated: 1057 g/mol. HPLC gradient Time (min) Flow %A %B (ml/min) 0,00 6,00 88,0 12,0 120,00 6,00 88,0 12,0 121,00 0,10 88,0 12,0 Example 1013 (General Procedure (Q)) 10 Benzotriazol-5-ylcarbonyl-Gly 2 -Arg 6
-NH
2
(BT-G
2
RF).
WO 2004/080480 PCT/DK2004/000158 391 HNy NH 2 HN yNH 2 HN yNH 2 NH NH NH N H H NN N NN H H
H
4 N N 0< QNz YNH HN HN HN
H
2 N NH H 2 N NH H 2 N 'NH MS (MALDI): m/z: 1214.8 g/mol; calculated: 1213.4 g/mol. HPLC gradient: Time (min) Flow %A %B (ml/min) 0,00 6,00 88,0 12,0 120,00 6,00 88,0 12,0 121,00 0,10 88,0 12,0 5 Example 1014 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-4-Abz-Gly 2 -Args-NH 2 (BT-4-Abz-G 2 Re). HNy NH 2 HNy NH 2 HN NH 2 NH NH NH 0 H O 0 H 0 H 0 N N N N N N NH2 N NI& H H 0 H 0 H 0 N HN HN H
H
2 N NH H 2 N NH MS (MALDI): m/z: 1176.7 g/mol; calculated: 1177.9 g/mol. 10 HPLC gradient: Time (min) Flow %A %B (ml/min) 0,00 6,00 95,0 5,0 40,00 6,00 60,0 40,0 45,00 6,00 60,0 40,0 50,00 6,00 0,0 100,0 55,00 6,00 0,0 100,0 WO 2004/080480 PCT/DK2004/000158 392 60,00 6,00 95,0 5,0 Example 1015 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-4-Abz-Gly-Args-NH 2 (BT-4-Abz-GR5). H HN~ NH 2 HN NH HN NH 2 N NH NH NH N H HN HN
H
2 N INH H 2 N NH 5 MS (MALDI): m/z: 1122 g/mol; calculated: 1120.4 g/mol. HPLC gradient: Time (min) Flow %A %B (ml/min) 0,00 6,00 95,0 5,0 40,00 6,00 60,0 40,0 45,00 6,00 60,0 40,0 50,00 6,00 0,0 100,0 55,00 6,00 0,0 100,0 60,00 6,00 95,0 5,0 Example 1016 (General Procedure (Q)) Benzotriazol-5-ylcarbonyl-4-Abz-Args-NH 2 (BT-4-Abz-Rs). H H 2 N NH H2N yNH NI H= N HN HN 0 N - H~ N N N NN N N-NH HHN N HNNH NNH NH 10 HN LNH2 HN" NH2 HN 2NH2 MS (MALDI): m/z: 1064.3 g/mol; calculated: 1063.2 g/mol.
WO 2004/080480 PCT/DK2004/000158 393 HPLC gradient: Time (min) Flow %A %B (ml/min) 0,00 6,00 95,0 5,0 40,00 6,00 60,0 40,0 45,00 6,00 60,0 40,0 50,00 6,00 0,0 100,0 55,00 6,00 0,0 100,0 60,00 6,00 95,0 5,0 General procedure (R) for preparation of compounds of general formula 117: H2N-PS H-(Arg);N-PS H H - (Gly)- (Arg)- N-PS H-(Abz)p (Gly)-(Arg); - PS 2 D HN 2 rE' B 1 " " N-(Abz)-(Gly)-(Arg)-N-PS O R 1 o OH m H X Y HNA
E
1 B2 D H NEB " N-(Abz);(Gly)m(Arg)KNH 2 O R 1 0 0H 117 5 wherein X, Y, R 1 0 , E, B', B 2 are as defined above, p is 0 or 1, m is 0-5 and n is 1-20. PS is a polymeric support, e.g. TentagenS RAM resin or a Rink amide resin. 10 WO 2004/080480 PCT/DK2004/000158 394 This general procedure is very similar to General procedure (Q), where benzotriazole-5 carboxylic acid in the last step before cleavage from the resin is replaced with compounds optionally prepared according to general procedure (D): Y HN EBB2 O 1 5,°B OH) O R 10 0 5 Example 1017 (General Procedure (R)) 4-{2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetylamino)benzoyl-Gly 2 -Arg-NH 2 O H 2 NYNH H 2 N YNH S H H HN. HN OHN H HN-^O N H 0 0 H 0 H 0 0,, N 'N N ;A H 0 0 H 0 H NH 2 N H NH NH HN INH 2 HN NH 2 HN NH 2 Example 1018 (General Procedure (R)) 10 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Args-NH 2 HNY NH 2 HN y NH 2 HNY NH 2 NH NH NH 0 0 H 0 S N ~ Y N Nj.k. NH1 2 HN H O H ON O 0 HN HN
H
2 N NH H 2 N NH MS (MALDI): m/z: 1057.3 g/mol; calculated: 1055.3 g/mol. Example 1019 (General Procedure (R)) 15 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloy-Arg 4
-NH
2 WO 2004/080480 PCT/DK2004/000158 395 HN yNH 2 HN NH 2 NH NH 00 0 H 0 s N N N N H2 HN N H O H O 0 HN HN
H
2 NilN H H 2 N INH MS (MALDI): m/z: 899.1 g/mol; calculated: 901.6 g/mol. Example 1020 (General Procedure (R)) 5 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg3-NH 2 HN.y NH 2 HN NH 2 NH NH 00 H 0 HN N H N H 0 0 HN
H
2 NH MS (MALDI): m/z: 746.2 g/mol; calculated: 742.9 g/mol. 10 Example 1021 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyry-Args-NH2
.
WO 2004/080480 PCT/DK2004/000158 396 HN yNH HN NH2 HN yNH 2 NH NH NH 0 0 H 0 H 0 O O /NO /NO N S N N" 'N N N N NH 2 HN I H O H O H O HN HN
H
2 N 'NH H 2 N -NH MS (MALDI): m/z: 1088.7 g/mol; calculated: 1087 g/mol. Example 1022 (General Procedure (R)) 5 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 4
-NH
2 . HNy NH 2 HN yNH 2 NH NH 0 o 0 0 -s ,,, , N:N N HNI1 HN H H 0 0 HN HN
H
2 N'- NH H 2 N /NH MS (MALDI): m/z: 933.0 g/mol; calculated: 931 g/mol. Example 1023 (General Procedure (R)) 10 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 3
-NH
2 HN y NH 2 HN yNH 2 NH NH 0 H 0 0 - N N"N
NH
2 HN J NHO HN H H 0 0 HN H2N NH WO 2004/080480 PCT/DK2004/000158 397 MS (MALDI): m/z: 776.9 g/mol; calculated: 774.0 g/mol. Example 1024 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg 12 -NH2 HN NH 2 HNy NH 2 HNyNH 2 HNy NH 2 HN NH 2 HNNH2 NH NH NH NH NH NH H H 0 H 0 H 0 H 0 H 0 OsN N N N NNN N N -NH HN HO H H H 0 O H 0 EH 0 HN HN HN HN HN HN 5 HN NH H 2 N NH H,N 1NH H 2 N -NH H 2 N )NH H 2 N NH MS (MALDI): m/z: 2232.9.4 g/mol; calculated: 2230.3 g/mol. Example 1025 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 8
-NH
2 . HNy NH2 HN yNH 2 HN yNH 2 HN y NH 2 NH NH NH NH H 0 H 0 H 0 H 0 S ON NNN NNNH HN H H H 0
-H
0 HN HN HN HN 10 H2N1NH H 2 N1 NH H 2 NiNH H 2 N NH 10 2H2N NHNH MS (MALDI): m/z: 1607.4 g/mol; calculated: 1605.5 g/mol. Example 1026 (General Procedure (R)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Args-NH 2
.
WO 2004/080480 PCT/DK2004/000158 398 HN yNH 2 HN yNH 2 HN y NH 2 NH NH NH o 0 H 0 H 0 SO N NN NH 2 HN H H H HN HN
H
2 N NH H 2 N NH MS (MALDI): m/z: 1141.9 g/mol; calculated: 1137.4 g/mol. Example 1027 (General Procedure (R)) 5 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg 4
-NH
2 . HN yNH2 HN NH NH NH S0O N N )NNH 2 HN H OH HN HN
H
2 N NH H 2 N 1NH MS (MALDI): m/z: 985.4 g/mol; calculated: 981.2 g/mol. Example 1028 (General Procedure (R)) 10 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 3
-NH
2 . HN y NH 2 HN yNH 2 NH NH 0 0 S N N N NH 2 HN
H
0 E H 0 HN
H
2 N JNH WO 2004/080480 PCT/DK2004/000158 399 MS (MALDI): m/z: 828.5 g/mol; calculated: 825.0 g/mol. The following compounds were prepared according to the methodology described in general procedure (Q) and (R): 5 Example 1029 4-(2H-Tetrazol-5-yl)benzoyl-4-Abz-Gly 2 -Arg 5
-NH
2 HNyNH 2 HN NH 2 HN YNH 2 NH NH NH 0N N N N N N NH2 NN=N HN ANH HN NH MS (MALDI): m/z: 1203.8 g/mol; calculated: 1203.8 g/mol. 10 Example 1030 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg5-NH2 HNNN N HN H NH HN MNH2 HN HNH2 00 00 NH HN HN H2NNNNNH H HN -NH MS (MALDI): m/z: 1152.5 g/mol; calculated: 1149.3 g/mol. 15 Example 1031 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Arg8-NH 2 WO 2004/080480 PCT/DK2004/000158 400 H , NN ,NN HNyNH 2
HN,NH
2 HiN-<.NH 2 HN2NH, N /NH NH NH NH N N 9 N N &N Nl}'N MJhH, H H H H 0 0 - 0 - 0 HN HN HN HN H 2 N NH HN NH H 2 N NH H 2 N NH MS (MALDI): m/z: 1621.0 g/mol; calculated: 1617.5 g/mol. Example 1032 5 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg 2
-NH
2 HN N HN NH, HN NH 2 HN NH, HN NH 2 HN NH, HN NH, N NH NH NH NH NH NH S H H 0 H H "IN N ~NX$N>)1. N N-:)- kN 'NH
H
0 H 0 0 0 H 0 HN HN HN HN HN HN
H
2 N NH H 2 N NH H N INH H 2 N NH H N NH H 2 N -NH MS (MALDI): m/z: 2247.9 g/mol; calculated: 2242.3 g/mol. General procedure (S) for preparation of compounds of general formula 118: WO 2004/080480 PCT/DK2004/000158 401
H
2 N-PS H-(Arg);N-PS H H-(Lys)-(Arg);N-PS m H 9 CGr-Lnk- (Gly)-- (Lys) - (Arg)- N -PS CGr-Lnk-(Gly)- (Lys)--(Arg) N H 2 118 Wherein PS is polymeric support, a Rink Amide AM resin, n is 0 - 20, m is 0 - 20, with the proviso that n + m <_20, wherein (Gly)p is defined as in a broader sense as Frgl above, p is 0 - 5, and furthermore the Frg2 (the Lys and Arg residues) may comprise one or more neutral 5 amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser, and wherein CGr-Lnk-OH, CGr, and Lnk is as described above. The compounds of the invention of general formula (118) can be prepared by means of standard peptide chemistry (as e.g. in General Procedure Q or R), e.g. in 7 mmol scale, 10 using Fmoc strategy and HOAt or HOBt activated amino acids. The compounds prepared in the following examples according to General Procedure (S) were all isolated as the TFA salts. This procedure is further illustrated in the following: Typically, 10 gram (7 mmol) of Rink Amide AM resin (Novabiochem 200-400 mesh) with substitution 0,70 mmol/g was treated with NMP by shaking for 3 h or more -then treated with 15 NMP/Piperidine/DBU (80/20/2) for (30 - 60 min) x 2 followed by wash with NMP x 6 which renders the resin ready for coupling. Step wise coupling of Fmoc-Arginine (Fmoc-Arg(Pbf)-OH). 21 mmol (13.6 g) of Fmoc-L-Arg(Pbf)-OH (MultiSyn Tech Gmbh., Germany) was dissolved in NMP (80 mL) together with HOBt-hydrate (21 mmol, 3.21 g). DIC (21 mmol, 3.27 mL) was 20 added to the solution and the mixture kept for 2-5 min before it was added to the resin which was shaken for a minimum 3 h, the resin was washed with NMP (80 mL). After each coupling step a capping step was performed using 20 x molar excess of Acetic acid / HOBt/DIC (8.4 WO 2004/080480 PCT/DK2004/000158 402 g/21.4 g/ 21 mL) in NMP (80 mL) followed by washing with NMP (80 mL) x 4 and the coupling was checked with TNBS (no red colour) .The Fmoc group was deprotected with NMP/Piperidine/DBU (80/20/2) for (30 - 60 min) x 2 followed by wash with NMP. Then another Fmoc-L-Arg(Pbf)-OH group was coupled as described above. 5 This stepwise assembling of the arginine residues was repeated to give the wanted number of arginines on the resin. When more than 6 residues were added double couplings were performed on each coupling step, one coupling carried out for 3 h or more- the other overnight. When Lysine or Glycine were part of the synthesised molecules the same procedure as 10 described above was used changing Fmoc-L-Arg(Pbf)-OH to Fmoc-L-Lys(Boc)-OH or Fmoc Gly-OH, respectively. Coupling of Ligand (CGr-Lnk-OH) to the polymer supported amino acid chain. The attachment of the ligand was carried out after deprotection of Fmoc as described above followed by coupling with the ligand containing a carboxylic acid using HOAt /DIC for 15 activation using the same ratio as described above. Double couplings and capping as described above were performed. Cleavage from the resin. The resin was washed with DCM (80 mL) x 4 followed by diethyl ether (80 mL ) x 4. Subsequently the resin was dried well and then added to a 20 fold excess of a mixture of 20 TFA / Thioanisol / Ethanol (90/5/5) and the mixture was stirred at RT overnight, evaporated to almost dryness and the residue was poured into a 20 fold excess of cold diethyl ether. The mixture was stirred for 30 min, filtered and the precipitate was washed with ether twice and dried. The dried compound was then dissolved in water and freeze dried, and the freeze dried material was then purified by HPLC. 25 RP-HPLC purification: The crude products were purified on RP-HPLC Kromasil RP18 100A 50x350 mm, flow 100mL/min; gradient: 0.0-2.0 min: 20% acetonotrile,0.1 % TFA; 2.0 17.0 min: 20 % acetonitrile, ,0.1 % TFA to 28 % acetonitrile, ,0.1 % TFA; 17.0-25.0 100 % acetoni trile, 0.1% TFA. Alternatively other HPLC systems were used. The identity was verified by mass spectrometry (MALDI-TOF). 30 The following examples were prepared using this general procedure. Example 1033 (General Procedure (S)) 5-[6-(5-Cyano-1lH-[1,2,3]triazol-4-yl)naphthalen-2-yloxy]pentanoyl-Arg 12
-NH
2 WO 2004/080480 PCT/DK2004/000158 403 HN NH HzN NH HN NH H 2 N NH HzN -NH H 2 N .NH NH NH NH NH NH NH HO H O HO- O H H ." ,, : H H H 6 NJ N- -H .HN H ;.,HN.0 ,N.H.0 N. .N , NH NH NH NH NH NH
H
2 N NH H 2 N 1NH H 2 N NH H 2 N NH H 2 N NH HN NH MS (MALDI-TOF): m/z: 2210 g/mol; calculated: 2209.2 g/mol. Example 1034 (General Procedure (S)) 5 4'-[5-(2H-Tetrazol-5-yl)indol-1 -ylmethyl]biphenyl-4-carbonyl-Arg 12
-NH
2 N, HN' 'N N H2N NH HN -NH H2N yNH H2N NH H2N -NH H2N NH NH NH NH NH NH NH I0 / NNN.N<NNN NN NNNH2 0 O H O H O H O H O H O NH NH NH NH NH NH
H
2 N iNH H 2 N "-NH H 2 N 'NH H 2 N)" NH H 2 N 'NH HN )NH MS (MALDI-TOF): m/z: 2268 g/mol; calculated: 2269 g/mol. Example 1035 (General Procedure (S)) 10 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argl4-NH 2
N
N N N' / HNy NH2 HNy NH, HNyNH 2 HNy NH 2 HNY NH 2 HN NH, HN NH 2 N NH NH NH NH NH NH NH '-"" N.'""H NH HN ./". N N N N _N "N N N HNN N N N N NH 4 N< J 4 JN HN4OJNNJN12 H H H 0 HO H H O H O HN HN HN- HN - HN HN- HN
H
2 N NH H 2 N NH HN NH H2N NH H 2 N NH H 2 N NH H 2 N NH MS (MALDI-TOF): m/z: 2553 g/mol; calculated: 2554 g/mol. Example 1036 (General Procedure (S)) 15 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys 2 -Arg 12
-NH
2 WO 2004/080480 PCT/DK2004/000158 404
.N,"
N HNN NNH, NH, NH, NH NH, NH, I \ N NH N NH N) NH N' NH NI NH NkNH NH NH NH NH N H H
N
H
N
H H N"N H H N N I N N N HN HN H N MS (MALDI-TOF): m/z: 2498 g/mol; calculated: 2499 g/mol. Example 1037 (General Procedure (S)) 50 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Gly 4 Arg NH 2 N2 M2lH f HN NH N NH HN NH MN NNH HN M MS (MALDI-TOF): m/z: 1845 g/mol; calculated: 1846 g/moL. HINNH HN N H 2 N yNH H 2 N yNH H,N NH H 2N yNH HN HN HN HN 0 H 0 H 0 H ' - H O H ) ' .'N N N N.J N .N N _.N N N NQNH 2 HH O H H H H- 0 0 A 0 0 0 0
HN
/
HN
/
HN
/ HN H2N N H H2N "'NH H"N NH HN i-lNH MS (MALDI-TOF): m/z: 1845 g/mol; calculated: 1846 g/mol. Example 1038 (General Procedure (S)) 10 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyI-Arg2 HN . NH H 2N yNH HN y NH H2N., NH HN N H H 2 N yNH
N
-
N H N' H N ,,H O N HN , H N NHN" HN, H H O ')
"
r A ' N HN H H )/ H N H 2N NH HN )IINH H,N 11,NH H 2N Ik NH H 2N J"NH H,N'I,'NH MS (MALDI-TOF): m/z: 2242 g/moI; calculated: 2243 g/mol.
WO 2004/080480 PCT/DK2004/000158 405 Example 1039 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Gly 2 -Arg 2 HqN ',N N. H N WN SNH N NH H 2 N NH H 2 NNH HNNH HN NH N HHN HN HN HN NH NH N X N N & NjN NJ*J~ &. "Y( U-H : J J NkH H O H O O H H 0 H O H O HN HN'f HNr HN HN)Z HN
NH
2 HN I-NH H 2 N 'NH HN NH H 2 N NH H 2 N NH HN NH MS (MALDI-TOF): m/z: 2497.5 g/mol; calculated: 2496 g/mol. 5 Example 1040 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Arg 1 6 NH 2 H-N H H NN NH HNHHN HNHHN NH2 HN NH 2 HN NH NH HNN HN HZyH y H y NI~NH N h HN HN HN HN NH NH NH NH 0II H ~H H ( 0 / N N _~N NN NN N N N N NNH -0 H0 H 0
H
0 H H O H 0 H HO~ HN N HNJ HNf HN HN HN' HN
H
2 N NH H 2 N NH H 2 N NH HzN NH H 2 N NH H 2 N lNH H 2 N NH HzN NH 10 MS (MALDI-TOF): m/z: 2873 g/mol; calculated: 2864 glmol. Example 1041 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lys 2 NH 2 15 NH 2 NH2 NH 2
NH
2
NH
2 MS (MALDI-TOF): m/z: 1905 g/mol; calculated: 1904 g/mol. N -' \1H 2
N
2 HN N 0
H
0
H
0
H
0 H H 0 4H Nv N.)y N.)y . YN~JiN N..)I.N N..'A.N N.)
-
0 EH 0 0 0EH 0 6 NH2 15 NH, NH 2
NH
2
NH
2
NH
2
NH
2 MS (MALDI-TOF): m/z: 1905 g/mol; calculated: 1904 g/mol.
WO 2004/080480 PCT/DK2004/000158 406 Example 1042 (General Procedure (S)) 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg ,NN HN NH H2 N NH H2N NH HN, f Y T N /HN HN HN H 0 H 0 H 0 N A N~ N N-AN N-JL.NH2 N)-Y .NY .. Y H H 0 H 0 H 0 HN HN HN
NH
2
H
2 N NH H 2 N NH H 2 N INH MS (MALDI-TOF): m/z: 1303 g/mol; calculated: 1304 g/mol. 5 Example 1043 (General Procedure (S)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 6 HNY NH 2 HN YNH 2 HN YNH 2 NH NH NH 00 H 0 H 0 H0 S & O N N N N N N NH2 H!- 2 HN H O H O H O 0 HN HN 'r HN
NH
2 H2N NNH H2N NH H2N NH NH2 MS (MALDI-TOF): m/z: 1293 g/mol; calculated: 1292 g/mol. 10 Other preferred compounds of the invention that may be prepared according to general pro cedures (Q), (R) and / or general procedure (S) include: Building block from example 469: 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyry-Arg, 0
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Argq-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-ArgrNH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Argl-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyryl-Arg 13
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyry-Arg 14
-NH
2 WO 2004/080480 PCT/DK2004/000158 407 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 15
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-ArgI 6
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 7-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Argj 8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Argas-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Arg 2 o-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 6
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryI-Lys 5
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 4
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 3
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 7
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysjo-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys, 1
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 12
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysj 3
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 4
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysj 5
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysl 6
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 1 7
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysi 8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lysjo-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Lys 2 0-NH 2 Building block from example 470: 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 6
-NH
2 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 5
-NH
2 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 4
-NH
2 5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 3
-NH
2 Building block from page 232: 6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]hexanoyl-Arg 3
-NH
2 6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]hexanoy-Arg 4
-NH
2 6-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]hexanoyl-Arg 5
-NH
2 Building block from page 232: 7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]heptanoyl-Arg 3
-NH
2 WO 2004/080480 PCT/DK2004/000158 408 7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]heptanoyl-Arg 4
-NH
2 7-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]heptanoyl-Args-NH 2 Building block from page 232: 8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]octanoyl-Arg 3
-NH
2 8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]octanoyl-Arg 4
-NH
2 8-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]octanoyl-Arg9 5
-NH
2 Building block from page 232: 10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoyl-Arg 3
-NH
2 10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoyl-Arg 4
-NH
2 10-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]decanoy-Args-NH 2 Building block from page 232: 11-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]undecanoy-Arg 3
-NH
2 11-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]undecanoyl-Arg 4
-NH
2 11-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]undecanoyl-Arg 5
-NH
2 Building block from page 232: 12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]dodecanoyl-Arg 3
-NH
2 12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]dodecanoyl-Arg 4
-NH
2 12-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]dodecanoyl-Arg 5
-NH
2 Building block from page 233: 15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentadecanoyl-Arg 3
-NH
2 15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentadecanoy-Arg 4
-NH
2 15-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentadecanoyl-Args-NH 2 Building block from example 283: 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Argo-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg 9
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyry-Arg 8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg 7
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg 6
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Arg 5
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-I -yloxy]butyryl-Arg 4
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Arg 3
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-I -yloxy]butyryl-Arg -NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Arg 12
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyry-Arg 13
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyry-Arg 14
-NH
2 WO 2004/080480 PCT/DK2004/000158 409 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Argj 5
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Arg 1 6-N H 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Arg 17
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Argr 8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Argy,-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Arg 2 0-N H 2 4-[4-(2,4-Dioxothiazolid in-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lys 6 -N H 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen- 1 -yloxy]butyryl-Lys 5
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lys 4
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lys 3 -N H 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lys 7
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lys8
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lysg-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyry-Lysio-NH2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]butyryl-Lysli-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lysl 2
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lys 1 a-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lysl 4
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lys 15
-NH
2 4-(4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lys1 6
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyry-Lys 17
-NH
2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lyss-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxy]butyryl-Lysy,-NH 2 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1 -yloxylbutyryl-Lys 2 o-NH 2 Building block from example 476: 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]acetyl-Arg 6
-NH
2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1-yloxy]acetyl-Arg 5
-NH
2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1 -yloxy]acetyl-Arg 4
-NH
2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1 -yloxy]acetyl-Arg 3
-NH
2 Building block from example 480: 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3 yl}acetyl-Arg 6
-NH
2 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3 yI}acetyl-Arg 5
-NH
2 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3- WO 2004/080480 PCT/DK2004/000158 410 yllacetyl-Arg 4
-NH
2 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylidene]-4-oxo-2-thioxothiazolidin-3 yI}acetyl-Arg 3
-NH
2 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg-NH 2 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg-NH 2 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg 4
-NH
2 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyryl-Arg 3
-NH
2 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg-NH 2 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg-NH 2 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg 4
-NH
2 15-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]pentadecanoyl-Arg 3
-NH
2 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoy-Arg 6 NH 2 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 5 NH 2 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg 4 NH 2 5-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]pentanoyl-Arg3
NH
2 Building block from example 462: 3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-Arg 6
-NH
2 Building block from example 473: 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg-NH 2 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg-NH 2 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg 4
-NH
2 2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg-NH 2 8-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]octanoyl-Arg-NH 2 8-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]octanoyl-Args-NH 2 8-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]octanoyl-Arg 4
-NH
2 8-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]octanoyl-Arg 3 -N H 2 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoyl-Arg-NH 2 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoyl-Arg-NH 2 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoyl-Arg 4
-NH
2 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]hexanoyl-Arg 3
-NH
2 Building block from example 466: WO 2004/080480 PCT/DK2004/000158 411 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 6
-NH
2 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Args-NH 2 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 4
-NH
2 4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 3
-NH
2 Building block from example 460: 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 6
-NH
2 Building block from example 467: 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg-NH 2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Args-NH 2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 4
-NH
2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-Arg 3
-NH
2 11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg r
-NH
2 11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Args-NH 2 11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg 4
-NH
2 11-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoyl-Arg 3
-NH
2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen- 1 -yloxy]butyryl-Arg 6
-NH
2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyryl-Args-NH 2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyry-Arg 4
-NH
2 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1 -yloxy]butyry-Arg 3
-NH
2 Building block from example 464: 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoy-Arg 6
-NH
2 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg-NH 2 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg 4
-NH
2 4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzoyl-Arg 3
-NH
2 Building block from example 463: 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-ArgB-NH 2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Args-NH 2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg 4
-NH
2 2-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg-NH 2 Building block from example 461: 2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-ArgB-NH 2 2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Args-NH 2 2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg 4
-NH
2 2-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-Arg 3
-NH
2 Building block from example 474: WO 2004/080480 PCT/DK2004/000158 412 4-3(,-ixtizldn5yieeehlpeoybtrlAg-H 4-3(,-ixtizldn5yienmty~hnx~uyy-rr-H 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethy)phenoxybutyry-Arg 4
-NH
2 4-3(,-ixtizldn5yieeehlpeoybtrlAg-H Building block from example 468: 4-2Boo4(-x--hoohaoii-5yieeehlpeoybtrlAg-H 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)pheoxy] butyryl-Arg 5
-NH
2 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)pheoxy]butyry-Arg 4 -NH2 4-2Boo-4oo2tixtizldn5yieeehlpeoybtrlAg-H Building block from example 738: 4-[3-(2H-Tetrazol-5-yl)carbazo-9-ylmethyl]benzoy-Arg 4 -NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Arg-NH 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]beflzoy-Argr-NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg-NH2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argo-NH 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg, ,-NH 2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyllbenzoyl-Arg 1 3 -N H 2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyl]benzoyl-Arglm-N H 2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyl]benzoyl-Arg 7 -N H 2 4-[3-(2H-Tetrazol-5-y)carbazol-9-ylmethylbenzoy-Arg 8 -NH2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyl]belzoy-Arg 9 -N H 2 4-[3-(2H-Tetrazol-5-y)carbazol-9-ylmethyl]benzoy-Arg 2 o-NH 2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyl]benzoyl-Lys 6 -N H 2 4-[3-(2H-Tetrazol-5-y)carbazol-9-ylmethylbenzoyl-LyS 4
-NH
2 4-[3-(2H-Tetrazol-5-y)carbazol-9-ylmethylbelzoy-LYS 3 -N H 2 4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylmethyl]benzoyl-LyS 7 -N H 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethylbenzoy-Lys 8 -N H 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lyso-N H 2 4-II3-(2H-TetrazoI-5-yI)carbazol-9-ylmethylbelzoyl-Lyslo-N H 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lysli -NH 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lysl 3 -N H 2 4-[3-(2H-Tetrazol-5-yl)carbazo-9-ylmethyl]bezoy-Lysl 4
-NH
2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethylbenzoyl-Lys-N H 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lysio-N H 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl] benzoyl-Lysi-N H 2 WO 2004/080480 PCT/DK2004/000158 413 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lysla-NH 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Lys 1 lg-NH 2 4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Lys 20
-NH
2 Building block from page 322: 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg-NH 2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg-NH 2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg 4
-NH
2 4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carbonyl-Arg 3
-NH
2 Building block from example 743: 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg 6
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Args-NH 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg 4
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg 3 -N H 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg-NH 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Arg 8
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argg-NH 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg 0
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argil-NH 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoy-Argl 2
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Arg, 3
-NH
2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argl 4 -N H 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argis-NH 2 3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-Argi 6
-NH
2 Example 1044 Equilibrium Solubility. For pH-solubility profiles, a 0.6 mM insulin stock solution containing 0.3 mM Zn 2 , 30 mM phenol, 1.6% glycerol and Zn 2 -binding ligand as required were prepared 5 and the pH was adjusted to the desired value corresponding to the alkaline endpoint of the pH-solubility profile. From these stock solutions samples were withdrawn, the pH adjusted to the desired value in the pH 3-8 range, and samples were incubated at 23 C for 24 hours. Af ter centrifugation (20,000 g in 20 min at 23 °C) of each sample, pH was measured and the solubility was determined by quantification of insulin contents in the supernatant by SEC 10 HPLC analysis WO 2004/080480 PCT/DK2004/000158 414 The effect of various concentration of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmrnethyl]benzoy-Arg 12
-NH
2 (Example 1032) on the pH-dependence of Gly 21 , AspB 2 8 insulin solubility is illustrated in Figure 1. 5 Example 1045 The effect of a high concentration of the ligand 4-[3-(2H-tetrazol-5-yl)carbazol-9 ylmethyl]benzoyl-Arg l 2
-NH
2 (Example 1032)on the pH-dependence of Gly" 1 insulin solubility is illustrated in Figure 2. The solubility was determined as in example 1044. Solution condi tions: 0.6 mM human insulin, 0.3 mM mM Zn 2 +, 30 mM phenol, 1.6% glycerol, 23 °C. 10 Example 1046 The slow release (prolonged action) properties of certain formulations of the present inven tion was characterized by the disappearance rate from the subcutaneous depot following 15 subcutaneous injections in pigs. To 50 % is the time when 50% of the A14 Tyr(1251) insulin has disappeared from the site of injection as measured with an external y-counter (Ribel et al., The Pig as a Model for Subcutaneous Absorption in Man. In: M. Serrano-Rtios and P.J. Le febre (Eds): Diabetes (1985) Proceedings of the 12 th congress of the International Diabetes Federation, Madrid, Spain, 1985 (Excerpta Medica, Amsterdam (1986), 891-896). The com 20 position of a series of protracted formulations is given in the table below together with the
T
50 o% values. The disappearance curves are illustrated in Figure 3 a-c, e-f. For comparison, the T 50 % for the corresponding insulin preparations formulated without the ligands would be about 2 hours. The disappearance curve for B29-N'-myristoyl-des(B30) human insulin (Fig ure 3 d) is 11 hours. 25 The induction of slow release by addition of exogenous ligands of the invention affords fur ther advantages in terms of versatility regarding the choice of insulin species and release patterns. Consequently, human or mutant insulins such as Gly A 2 ', Gly"lAspB 28 may be for mulated as slow- or dual-release preparations by adding variable amounts of HisBo Zn2+-site 30 ligand. This is illustrated below for Gly" 1 , Asp B28 human insulin and Gly" human insulin em ploying different Zn2+-site ligands. As shown in the table below and in Figure 3 panels a-c and e-f, addition of ligand produces a slow release preparation with T0o% in the range of 5 to 16 hours.
WO 2004/080480 PCT/DK2004/000158 415 2I5-Prep. 1 "I--Prep. 2 1 "I-Prep. 3 z"I-Prep. 4 'Pr -Prep. 6 0.6 B29-N' 0.6 GlyA2, 0.6 Gly" , 0.6Gl 1 06 ly 1 , 0.6 Gly 1, myristoyl- 2 Insulin Asp
B
28 AspB 2 8 Asp0.6 Gy 2 , myristoyl-(B30) 0.6 Gly"' 0.6 Gly 1 (mM) human insu- human in- human insulin human insulin insulin human insulin human lin sulin insulin Zn 2 + (mM) 0.3 0.3 0.3 0.22 0.3 0.3 16mM Phenolic 30mM phe- 30mM phenol, 30 mM 30 mM 30mM phenol ligand nol phenol 16mM m- phenol phenol cresol 0.6 mM 1.2 mM 6mM 6mM 2mM 6mM 6mM 2mM 1430R 12 1430R 12 1253R3 1253Re 1081AbzG2Rs Zn 2 + ligand 1253R 1253R 5 1081AbzG 2
R
5 20 mM NaCI (Example (Example (Example (Example (Example missing, missing, missing, missing, 781) 779) 782) PeM) PeM) Glycerol (%) 1.6 1.6 1.6 1.6 1.6 1.6 Phosphate 0 0 0 7 0 0 buffer (mM) pH 4.0 4.0 4.0 7.5 4.0 4.0
T
50 o% (hrs) 9.3 8.7 5.3 11.0 14.4 16.5 Figure 3 shows the disappearance from the subcutaneous depot (pig model) of insulin prepa rations. Curves a)-c) are GlyA 2 , Asp B28 human insulin formulated with an excess concentra 5 tion compared to Zn 2 + of a) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 yloxy]butyryl-Arg3-NH 2 , b) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1 yloxy]butyryl-Args-NH 2 and c) 4-(2H-Tetrazol-5-yl)benzoyl-Abz-Gly2-Args-NH2. Curve d) is B29-N'-myristoyl-des(B30) human insulin. Curves e) and f) are Gly
A
' human insulin formu lated with two different excess concentrations compared to Zn 2 + of 4-[3-(2H-Tetrazol-5 10 yl)carbazol-9-ylmethyl]benzoy-Arg1 2
-NH
2
.
WO 2004/080480 PCT/DK2004/000158 416 ANALYTICAL METHODS Assays to quantify the binding affinity of ligands to the metal site of the insulin R 6 hexamers: 5 4H3N-assay: The binding affinity of ligands to the metal site of insulin R6 hexamers are measured in a UV/vis based displacement assay. The UV/vis spectrum of 3-hydroxy-4-nitro benzoic acid (4H3N) which is a known ligand for the metal site of insulin R6 shows a shift in absorption 10 maximum upon displacement from the metal site to the solution (Huang et al., 1997, Bio chemistry 36, 9878-9888). Titration of a ligand to a solution of insulin R 0 hexamers with 4H3N mounted in the metal site allows the binding affinity of these ligands to be determined follow ing the reduction of absorption at 444 nm. A stock solution with the following composition 0.2 mM human insulin, 0.067 mM Zn-acetate, 15 40 mM phenol, 0.101 mM 4H3N is prepared in a 10mL quantum as described below. Buffer is always 50mM tris buffer adjusted to pH=8.0 with NaOH/CIO4. 1000 gL of 2.0mM human insulin in buffer 20 66.7 pL of 10mM Zn-acetate in buffer 800 gL of 500mM phenol in H 2 0 201 iL of 4H3N in H 2 0 7.93 ml buffer 25 The ligand is dissolved in DMSO to a concentration of 20 mM. The ligand solution is titrated to a cuvette containing 2 mL stock solution and after each addi tion the UV/vis spectrum is measured. The titration points are listed in Table 3 below. 30 WO 2004/080480 PCT/DK2004/000158 417 Table 3 ligand ligand addition cone. dilution (pl) (mM) factor 1 0.010 1.0005 1 0.020 1.0010 1 0.030 1.0015 2 0.050 1.0025 5 0.100 1.0050 10 0.198 1.0100 20 0.392 1.0200 20 0.583 1.0300 20 0.769 1.0400 20 0.952 1.0500 5 The UV/vis spectra resulting from a titration of the compound 3-hydroxy-2-naphthoic acid is shown in Figure 4. Inserted in the upper right corner is the absorbance at 444nm vs. the con centration of ligand. The following equation is fitted to these datapoints to determine the two parameters KD(obs), 10 the observed dissociation constant, and absmx the absorbance at maximal ligand concentra tion. abs ([ligand]free) = (absmax * [ligand]free)/ (KD(obs) + [ligand]free) 15 The observed dissociation constant is recalculated to obtain the apparent dissociation con stant KD(app) = KD(obs) / ( 1+[4H3N]/K 4 H3N) 20 The value of K 4
H
3 N=50 pM is taken from Huang et al., 1997, Biochemistry 36, 9878-9888. TZD-assay: The binding affinity of ligands to the metal site of insulin R 6 hexamers are measured in a fluo rescense based displacement assay. The fluorescence of 5-(4- WO 2004/080480 PCT/DK2004/000158 418 dimethylaminobenzylidene)thiazolidine-2,4-dione (TZD) which is a ligand for the metal site of insulin R 6 is quenched upon displacement from the metal site to the solution. Titration of a ligand to a stock solution of insulin Re hexamers with this compound mounted in the metal site allows the binding affinity of these ligands to be determined measuring the fluorescence 5 at 455nm upon excitation at 410 Onm. Preparation Stock solution: 0.02 mM human insulin, 0.007 mM Zn-acetate, 40 mM phenol, 0.01 mM TZD in 50mM tris buffer adjusted to pH=8.0 with NaOH/CIO4". 10 The ligand is dissolved in DMSO to a concentration of 5 mM and added in aliquots to the stock solution to final concentrations of 0-250 pM. Measurements Fluorescence measurements were carried out on a Perkin Elmer Spectrofluorometer 15 LS50B.The main absorption band was excited at 410 nm and emission was detected at 455 nm. The resolution was 10 nm and 2.5 nm for excitation and emission, respectively. The fluorescence spectra resulting from a titration of the compound 5-(4 dimethylaminobenzylidene)thiazolidine-2,4-dione (TZD) is shown in Figure 5. Inserted in the 20 upper right corner is the fluorescence at 455 nm upon exitation at 410 nM vs. the concentra tion of ligand. Data analysis This equation is fitted to the datapoints 25 AF(455nm)) = AFmax * [ligand]ree/( KD(app) * ( 1+[TZD]/KTZD)+ [ligand]free)) KD(app) is the apparent dissociation constant and Fmax is the fluorescence at maximal ligand concentration. The value of KTZD is measured separately to 230 nM Two different fitting-procedures can be used. One in which both parameters, KD(app) and 30 Fmax, are adjusted to best fit the data and a second in which the value of Fmax is fixed (Fmax=1) and only KD(app) is adjusted. The given data are from the second fitting procedure. The Solver module of Microsoft Excel can be used to generate the fits from the datapoints.

Claims (233)

1. A pharmaceutical preparation comprising
4. Acid-stabilised insulin
5. Zinc ions 5 6. A zinc-binding ligand of the following general formula (I) CGr-Lnk-Frg1-Frg2-X (I) wherein: 10 CGr is a chemical group which reversibly binds to a HisBio Zn 2+ site of an insulin hexamer; Lnk is a linker selected from * a valence bond 15 * a chemical group GB of the formula -B 1 -B 2 -C(O)-, -B 1 -B 2 -SO 2 -, -B 1 -B 2 -CH 2 -, or -B 1 B 2 -NH-; wherein B 1 is a valence bond, -0-, -S-, or -NR 6B -, B 2 is a valence bond, C 1 -C 18 -alkylene, C 2 -C 1 8 -alkenylene, C 2 -C 1 8 -alkynylene, arylene, heteroarylene, -C 1 -Cl 8 -alkyl-aryl-, -C 2 -C 1 8 -alkenyl-aryl-, -C 2 -C 1 8 -alkynyl-aryl-, -C(=O) C 1 -C 1 8 -alkyl-C(=O)-, -C(=O)-C 1 -C 18 -alkenyl-C(=O)-, -C(=O)-Cj-C 1 8 -alkyl-O-C-Cs 8 20 alkyl-C(=O)-, -C(=O)- C 1 -C, 8 -alkyl-S-C 1 -C 18 -alkyl-C(=O)-, -C(=O)-C 1 -C, 8 -alkyl-NR 6 -Cj C 1 8 -alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C(=O)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF 3 , -OCF 3 , -OR 6 B, or -NReBR 7 B and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR 6B , -C(O)H, OCOR 6 8 , -SO 2 , -CN, -CF 3 , 25 -OCF 3 , -NO 2 , -OR 6 8 , -NR 6 "R 7 8 , C 1 -C18-alkyl, or C 1 -C 1 a-alkanoyl; R 68 and R 7B are independently H, C 1 -C 4 -alkyl; Frgl is a fragment consisting of 0 to 5 neutral a- or 3-amino acids 30 Frg2 is a fragment comprising 1 to 20 positively charged groups independently selected from amino or guanidino groups; and X is -OH, -NH 2 or a diamino group, WO 2004/080480 PCT/DK2004/000158 420 or a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric forms. 2. A pharmaceutical preparation according to claim 1 wherein CGr is a chemical structure 5 selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophe nolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimi dazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5-mercaptotetrazoles, rho danines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids. 10 3. A pharmaceutical preparation according to claim 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4-cyano-1,2,3-triazoles. 15 4. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is X H Y 0Y 0 N 0 A HN or HN NB orR 3 A O R O 4 R1 A RO RA 0 R0 2 0 wherein X is =0, =S or =NH Y is -S-, -0- or -NH 20 R 1 , R 1 A and R 4 are independently selected from hydrogen or Cl-C 6 -alkyl, R 2 and R" are hydrogen or Cl-C 6 -alkyl or aryl, R' and R 2 may optionally be combined to form a double bond, RIA and R 2A may optionally be combined to form a double bond, R 3 , R 3 A and R 5 are independently selected from hydrogen, halogen, aryl optionally substi 25 tuted with one or more substituents independently selected from R' 6 , Cl-Co-alkyl, or -C(0)NR"R 12 , A, A' and B are independently selected from C 1 -C 6 -alkyl, aryl, aryl-Cl-C 6 -alkyl, -NR"-aryl, aryl-C 2 -C 6 -alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one 30 or more substituents independently selected from R 6 and the aryl or heteroaryl is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 , WO 2004/080480 PCT/DK2004/000158 421 A and R 3 may be connected through one or two valence bonds, B and R 5 may be connected through one or two valence bonds, R" is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 , R 7 , R 8 , R 9 and R 1 0 are independently selected from 5 *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -OS(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 11 , -NR 11 R 12 , -SR 1 1 , -NR"S(O) 2 R 1 2 , -S(O) 2 NR"R 1 2 , -S(O)NR"R 1 2 , -S(O)R'", -S(O) 2 R 11 , -OS(O) 2 R 11 , -C(O)NR"R 12 , -OC(O)NR"R 12 , -NR 11 C(O)R 12 , -CH 2 C(O)NR"R 1 2 10 -OC-C 6 -alkyl-C(O)NR 1 1 R 12 , -CH 2 0R 11 , -CH 2 0C(O)R 1 , -CH 2 NR" R 12 , -OC(O)R", -OCI-C 15 -alkyl-C(O)OR 1 , -OCI-C 6 -alkyl-OR 11 , -SC 1 -C 6 -alkyl-C(O)OR 1 -C 2 -C 6 -alkenyl-C(=O)OR 1 1 , -NR"-C(=O)-Cl-C 6 -alkyl-C(=O)OR 1 , -NR 1 -C(=O)-Cl-C 6 -alkenyl-C(=O)OR" 1 , -C(O)OR" 11 , C(O)R 11 , or -C 2 -C 6 -alkenyl C(=O)R", =0, or -C 2 -C 6 -alkenyl-C(=O)-NR" R 12 , 15 * Cl-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, each of which may optionally be substi tuted with one or more substituents independently selected from R 1 3 , * aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, 20 aryl-C 2 -Ce-alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cl C 6 -alkyl, heteroaryl-C 2 -C 6 -alkenyl, heteroaryl-C 2 -C 6 -alkynyl, or C3-C6 cycloalkyl, of which each cyclic moiety may optionally be substituted with one or more substitu ents independently selected from R 14 25 R 11 and R 12 are independently selected from hydrogen, OH, C l -C 2 0 -alkyl, aryl-CI-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 1 and R 12 when attached to the same 30 nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 1 3 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR", -C(O)OR" , -NR11R 2, 35 and -C(O)NR"R 12 WO 2004/080480 PCT/DK2004/000158 422 R 14 is independently selected from halogen, -C(O)OR 11 , -CH 2 C(O)OR 1 1 , -CH 2 OR 1 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR 11 , -NR 11 R 1 2 , -NR 11 C(O)R 11 , -S(O) 2 R 1 , aryl and Cl-C 6 -alkyl, 5 R 15 is independently selected from halogen, -CN, -CF 3 , =0, -OCF 3 , -OC 1 -C 6 -alkyl, -C(O)OC, Ce-alkyl, -COOH and -NH 2 , R 16 is independently selected from halogen, -C(O)OC,-C 6 -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , NO 2 , -OH, -OC 1 -C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, or any enantiomer, diastereomer, in 10 cluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically accept able acid or base. 5. A pharmaceutical composition according to claim 4 wherein X is =0 or =S.
6. A pharmaceutical composition according to claim 5 wherein X is =0. 15 7. A pharmaceutical composition according to claim 5 wherein X is =S.
8. A pharmaceutical composition according to any one of the claims 4 to 7 wherein Y is -0 or -S-.
9. A pharmaceutical composition according to claim 8 wherein Y is -0-.
10. A pharmaceutical composition according to claim 8 wherein Y is -NH-. 20 11. A pharmaceutical composition according to claim 8 wherein Y is -S-.
12. A pharmaceutical composition according to any one of the claims 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
13. A pharmaceutical composition according to claim 12 wherein A is selected from ArG1 25 optionally substituted with up to four substituents, R 7 , Ra, R 9 , and R 1 0 which may be the same or different.
14. A pharmaceutical composition according to claim 13 wherein A is phenyl or naphtyl op tionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different. 30 15. A pharmaceutical composition according to claim 14 wherein A is 99 Ror R 9 or RR7 R WO 2004/080480 PCT/DK2004/000158 423
16. A pharmaceutical composition according to claim 14 wherein A is phenyl.
17. A pharmaceutical composition according to any one of the claims 4 to 11 wherein A is heteroaryl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or different. 5 18. A pharmaceutical composition according to claim 17 wherein A is selected from Heti op tionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 o which may be the same or different.
19. A pharmaceutical composition according to claim 18 wherein A is selected from Het2 op tionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or 10 different.
20. A pharmaceutical composition according to claim 19 wherein A is selected from Het3 op tionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 o which may be the same or different.
21. A pharmaceutical composition according to claim 20 wherein A is selected from the 15 group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl, wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 1 0 which may be the same or different.
22. A pharmaceutical composition according to claim 20 wherein A is benzofuranyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 which may be the same or differ 20 ent.
23. A pharmaceutical composition according to claim 22 wherein A is R 8 0R 8 R R 7 Oor or R7
24. A pharmaceutical composition according to claim 20 wherein A is carbazolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 1 0 which may be the same or differ 25 ent.
25. A pharmaceutical composition according to claim 24 wherein A is R 9 R8 H N R 7 WO 2004/080480 PCT/DK2004/000158 424
26. A pharmaceutical composition according to claim 20 wherein A is quinolyl optionally sub stituted with up to four substituents R 7 , R 8 , R 9 , and R 1 o which may be the same or different.
27. A pharmaceutical composition according to claim 26 wherein A is R 9 R 9 or 8 R7 R R R 5 28. A pharmaceutical composition according to claim 20 wherein A is indolyl optionally sub stituted with up to four substituents R 7 , R 8 , R 9 , and R 1 o which may be the same or different.
29. A pharmaceutical composition according to claim 28 wherein A is R8 R9 R NH or R N H
30. A pharmaceutical composition according to any one of the claims 4 to 29 wherein R 1 is 10 hydrogen.
31. A pharmaceutical composition according to any one of the claims 4 to 30 wherein R 2 is hydrogen.
32. A pharmaceutical composition according to any one of the claims 4 to 29 wherein R 1 and R 2 are combined to form a double bond. 15 33. A pharmaceutical composition according to any one of the claims 4 to 32 wherein R 3 is C,-CB-alkyl, halogen, or C(O)NR 16 R 17 .
34. A pharmaceutical composition according to claim 33 wherein R 3 is Cl-C 6 -alkyl or C(O)NR 6 R 17 .
35. A pharmaceutical composition according to claim 34 wherein R 3 is methyl. 20 36. A pharmaceutical composition according to any one of the claims 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and Rio which may be the same or different.
37. A pharmaceutical composition according to any one of the claims 4 to 11 or 36 wherein R 4 is hydrogen. 25 38. A pharmaceutical composition according to any one of the claims 4 to 11 or 36 to 37 wherein R 5 is hydrogen. WO 2004/080480 PCT/DK2004/000158 425
39. A pharmaceutical composition according to any one of the claims 4 to 38 wherein R 6 is aryl.
40. A pharmaceutical composition according to claim 39 wherein R 6 is phenyl.
41. A pharmaceutical composition according to any one of the claims 4 to 40 wherein R , R 8 , 5 R 9 and R 10 are independently selected from *hydrogen, halogen, -NO 2 , -OR 11 , -NR"R 1 2 , -SR 11 " , -NR"S(O) 2 R 1 2 , -S(O) 2 NR"R 12 , -S(O)NR"R 12 , -S(O)R 1 ", -S(O) 2 R 11 , -OS(0) 2 R 11 , -NR 1 1 C(O)R 12 , -CH 2 0R 1 , CH 2 OC(O)R 11 , -CH 2 NR" R 12 , -OC(O)R 11 , -OC 1 -C 6 -alkyl-C(O)OR' 1 , -OCI-C 10 alkyl-C(O)NR"R 12, -OC 1 -C 6 -alkyl-OR 11 , -SC 1 -Ce-alkyl-C(O)OR 1 , -C 2 -C 6 -alkenyl C(=O)OR 11 , -C(O)OR 1 1 , or -C 2 -C 6 -alkenyl-C(=O)R 11 , * C 1 -C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may each optionally be substituted with one or more substituents independently selected from R 1 3 15 * aryl, aryloxy, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-C 1 -C 6 -alkyl, aryl-C 2 Co-alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cl-Co-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from Ri. 20 42. A pharmaceutical composition according to claim 41 wherein R 7 , R 8 , R 9 and R W o are independently selected from *hydrogen, halogen, -NO 2 , -OR", -NR" 1 R' 12 , -SR", -S(O) 2 R", -OS(0)2 R 11 , CH 2 OC(O)R 11 , -OC(O)R 11 , -OC,-C 6 -alkyl-C(O)OR 1 , -OCj-C 6 -alkyl-OR 11, -SC-C6 25 alkyl-C(O)OR 1 , -C(0)OR", or -C 2 -C 6 -alkenyl-C(=O)R' 1 , * Cl-C 6 -alkyl or Cl-C 6 -alkenyl which may each optionally be substituted with one or more substituents independently selected from R 13 30 * aryl, aryloxy, aroyl, aryl-C l -C 6 -alkoxy, aryl-C,-C 6 -alkyl, heteroaryl, of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 1 4
43. A pharmaceutical composition according to claim 42 wherein R 7 , R 8 , R 9 and R W o are inde 35 pendently selected from WO 2004/080480 PCT/DK2004/000158 426 *hydrogen, halogen, -NO 2 , -OR 11 , -NR 1 "R 12 , -SR 11 , -S(O) 2 R 1 , -OS(O) 2 R 11 , CH 2 OC(O)R 11 , -OC(O)R 11 , -OCI-C 6 -alkyl-C(O)OR 1 , -OC 1 -C 6 -alkyl-OR 1 , -SC1-C 6 alkyl-C(O)OR 1 1 , -C(O)OR 11 , or -C 2 -C 6 -alkenyl-C(=O)R 1 , 5 * Cl-C 6 -alkyl or CI-C 6 - which may each optionally be substituted with one or more substituents independently selected from R 13 * aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, 10 of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14
44. A pharmaceutical composition according to claim 43 wherein R 7 , R 8 , R 9 and R 1 0 are independently selected from 15 * hydrogen, halogen, -OR 11 , -OC 1 -C 6 -alkyl-C(O)OR" 1 , or -C(O)OR 1 , * CI-C 6 -alkyl which may each optionally be substituted with one or more substituents independently selected from R 13 20 e aryl, aryloxy, aryl-Cl-C 6 -alkoxy, of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R".
45. A pharmaceutical composition according to claim 44 wherein R 7 , R 8 , R 9 and R 1 0 are 25 independently selected from *hydrogen, halogen, -OR 1 ", -OC 1 -C 6 -alkyl-C(O)OR" 1 , or -C(O)OR", * C 1 -C 6 -alkyl which may each optionally be substituted with one or more substituents independently selected from R 1 3 30 *ArG1, ArGloxy, ArG1-Cl-C 6 -alkoxy, of which each of the cyclic moieties optionally may be substituted with one or more substitu ents independently selected from R 14 WO 2004/080480 PCT/DK2004/000158 427
46. A pharmaceutical composition according to claim 45 wherein R 7 , R 8 , R 9 and R 1 0 are independently selected from *hydrogen, halogen, -OR 11 , -OCj-C 6 -alkyl-C(O)OR 1 , or -C(O)OR", 5 * Cl-C 6 -alkyl which may optionally be substituted with one or more substituents inde pendently selected from R 13 * phenyl, phenyloxy, phenyl-C 1 -C 6 -alkoxy, wherein each of the cyclic moieties option ally may be substituted with one or more substituents independently selected from 10 R 1 4
47. A pharmaceutical composition according to any one of the claims 4 to 46 wherein R 11 and R12 are independently selected from hydrogen, C 1 -C 20 -alkyl, aryl or aryl-Cl-C 6 -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substitu 15 ents independently selected from R 16 ; R" 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
48. A pharmaceutical composition according to claim 47 wherein R 11 and R 12 are independ 20 ently selected from hydrogen, C 1 -C 20 -alkyl, aryl or aryl-Cl-C 6 -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R' 6 .
49. A pharmaceutical composition according to claim 48 wherein R 1 " and R 1 2 are independ 25 ently selected from phenyl or phenyl-Cl-C 6 -alkyl.
50. A pharmaceutical composition according to claim 48 wherein one or both of R 1 " and R 12 are methyl.
51. A pharmaceutical composition according to any one of the claims 4 to 50 wherein R 3 is independently selected from halogen, CF 3 , OR" 1 or NR 11 R 1 2 . 30 52. A pharmaceutical composition according to claim 51 wherein R 13 is independently se lected from halogen or OR".
53. A pharmaceutical composition according to claim 52 wherein R 13 is OR".
54. A pharmaceutical composition according to any one of the claims 4 to 53 wherein R 1 4 is independently selected from halogen, -C(O)OR", -CN, -CF 3 , -OR" 1 , S(O) 2 R", and Cl 35 C 6 -alkyl. WO 2004/080480 PCT/DK2004/000158 428
55. A pharmaceutical composition according to claim 54 wherein R 1 4 is independently se lected from halogen, -C(O)OR 1 , or -OR 11 .
56. A pharmaceutical composition according to any one of the claims 4 to 55 wherein R 5 is is independently selected from halogen, -CN, -CF 3 , -C(O)OCI-C 6 -alkyl,and -COOH. 5 57. A pharmaceutical composition according to claim 56 wherein R 15. is independently se lected from halogen or -C(O)OC-C 6 -alkyl.
58. A pharmaceutical composition according to any one of the claims 4 to 57 wherein R 16 is independently selected from halogen, -C(O)OC 1 -Co-alkyl, -COOH, -NO 2 , -OC 1 -C 6 -alkyl, -NH 2 , C(=O) or C 1 -C 6 -alkyl. 10 59. A pharmaceutical composition according to claim 58 wherein R 16 is independently se lected from halogen, -C(O)OC 1 -C 6 -alkyl, -COOH, -NO 2 , or C 1 -C 6 -alkyl.
60. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is 17 -N \D N \ or NDE Or F G N 18 N N E orG H R H R 1 9 H O /R20 or N D1-G H 15 wherein R 19 is hydrogen or Cl-C 6 -alkyl, R 20 is hydrogen or Cl-C6-alkyl, D, D1 and F are a valence bond, Cl-C 6 -alkylene or C-C-alkenylene optionally substituted 20 with one or more substituents independently selected from R 72 R 7 2 is independently selected from hydroxy, C 1 -C 6 -alkyl, or aryl, E is Cl-C 6 -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with 25 up to three substituents R 21 , R2 and R 23 , G and G' are Cl-C 6 -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub stituted with up to three substituents R 24 , R 25 and R , R 17 , R 18 , R 21 , R2, R 23 , R 24 , R 25 and R 26 are independently selected from WO 2004/080480 PCT/DK2004/000158 429 *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(0) 2 CF 3 , -SCF 3 , -NO 2 , =0, -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 S(O) 2 R 28 , -S(O) 2 NR 27 R 28 , -S(O)NR 27 R 28 , -S(O)R 27 , -S(O) 2 R 27 , -C(O)NR 2 7 R 2 8 , -OC(O)NR 2 7 R 28 , 5 -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -CH 2 C(O)NR 2 7 R 28 , -OCH 2 C(O)NR 2 7 R 28 , -CH 2 0R 27 , -CH 2 NR 27 R 28 , -OC(O)R 7 , -OCI-C 6 -alkyl-C(O)OR 27 , -SC 1 -C 6 -alkyl-C(O)OR 27 , -C 2 -C alkenyl-C(=O)OR 27 , -N R 27 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 27 , -NR 27 C(=O)-C,-C 6 alkenyl-C(=O)OR 27 , -C(=O)NR 27 C 1 -C 6 -alkyl-C(=O)OR 2 7 , -C6-C 6 -alkyl-C(=O)OR 27 ,or -C(0)OR 2 7 10 * CI-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R", 15 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2 -C 6 alkenyl or heteroaryl-C 2 -C 6 -alkynyl, 20 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 R 27 and R 28 are independently selected from hydrogen, Cl-C 6 -alkyl, aryl-Cl-Ce-alkyl or aryl, or 25 R 27 and R 28 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 30 R" 9 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 27 , and -NR 27 R 28 R 30 is independently selected from halogen, -C(O)OR 27 , -CN, -CF3, -OCF 3 , -NO 2 , -OR 27 , -NR 27 R 28 and Cl-C 6 -alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. 35 61. A pharmaceutical composition according to claim 60 wherein D is a valence bond. WO 2004/080480 PCT/DK2004/000158 430
62. A pharmaceutical composition according to claim 60 wherein D is Cl-C 6 -alkylene option ally substituted with one or more hydroxy, Cl-C 6 -alkyl, or aryl.
63. A pharmaceutical composition according to any one of the claims 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three sub 5 stituents independently selected from R 2 1 , R 22 and R 23
64. A pharmaceutical composition according to claim 63 wherein E is aryl optionally substi tuted with up to three substituents independently selected from R 21 , R 22 and R 23
65. A pharmaceutical composition according to claim 64 wherein E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 10 and R 2 3
66. A pharmaceutical composition according to claim 65 wherein E is phenyl optionally sub stituted with up to three substituents independently selected from R 2 1 , R" and R 2 3
67. A pharmaceutical composition according to claim 66 wherein CGr is N. 19 R N H 15 68. A pharmaceutical composition according to any one of the claims 60 to 67 wherein R 2 1 , R 22 and R 23 are independently selected from *hydrogen, halogen, -CHF 2 , -CF3, -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -SCF 3 , NO 2 , -OR 27 , -NR 27 R 28 , -SR 27 , -C(O)NR 27 R 2 8 , -OC(O)NR 27 R 28 , -NR 27 C(O)R 28 , 20 -NR 27 C(O)OR 28 , -CH 2 C(O)NR 27 R 28 , -OCH 2 C(O)NR 27 R 28 , -CH 2 0R 27 , -CH 2 NR 27 R 28 , -OC(O)R 27 , -OC 1 -C 6 -alkyl-C(O)OR 2 7 , -SC 1 l-C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -N R 27 -C(=O)-Cl-C-alkyl-C(=O)OR 27 , -NR 27 -C(=O)-C 1 -C 6 alkenyl-C(=O)OR 27 -, -C(=O)NR 7 -Cl-C 6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR, 25 * Ci-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 29 30 WO 2004/080480 PCT/DK2004/000158 431 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C 2 -C 6 alkenyl or heteroaryl-C 2 -C 6 -alkynyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 .
69. A pharmaceutical composition according to claim 68 wherein R 2 1 , R 22 and R 23 are inde pendently selected from 10 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 2 8 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)N R 27 -C-C-alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 7 , 15 * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 29 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-C,-C 6 alkyl, 20 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30
70. A pharmaceutical composition according to claim 69 wherein R 21 , R 22 and R 23 are inde pendently selected from 25 * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27, -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)N R 27 -C-C-alkyl-C(=O)OR 27 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 27 30 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 Saryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 35 alkyl WO 2004/080480 PCT/DK2004/000158 432 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 .
71. A pharmaceutical composition according to claim 70 wherein R 2 1 , R 22 and R 23 are inde pendently selected from 5 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SCI-C 6 -alkyl-C(O)OR 2 7 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)NR 27 -C 1 -C 6 -alkyl-C(=O)OR 27 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 2 7 10 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 * ArG1, ArG1-O-, ArGI-C(O)-, ArGl-Cl-C 6 -alkoxy, ArGl-Cl-C 6 -alkyl, Het3, Het3-C 15 C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 .
72. A pharmaceutical composition according to claim 71 wherein R 2 1 , R 22 and R 23 are inde pendently selected from 20 * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OC,-Co-alkyl-C(O)OR 27 , -SC,-CB-alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)NR 27 C -C 6 -alkyl-C(=O)OR 27 , -C -C 6 -alkyl-C(=O)OR 2 7 , or -C(O)OR 27 25 * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 29 * phenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, phenyl-Cl-C 6 -alkyl, 30 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 9 .
73. A pharmaceutical composition according to any one of the claims 60 to 72 wherein R 1 9 is hydrogen or methyl.
74. A pharmaceutical composition according to claim 73 wherein R 1 9 is hydrogen. WO 2004/080480 PCT/DK2004/000158 433
75. A pharmaceutical composition according to any one of the claims 60 to 74 wherein R 2 7 is Hydrogen, C 1 -C 6 -alkyl or aryl.
76. A pharmaceutical composition according to claim 75 wherein R 27 is hydrogen or CI-C 6 alkyl. 5 77. A pharmaceutical composition according to any one of the claims 60 to 76 wherein R 28 is hydrogen or Cl-C 6 -alkyl.
78. A pharmaceutical composition according to claim 60 wherein F is a valence bond.
79. A pharmaceutical composition according to claim 60 wherein F is C 1 -C 6 -alkylene option ally substituted with one or more hydroxy, C 1 -C 6 -alkyl, or aryl. 10 80. A pharmaceutical composition according to any one of the claims 60 or 78 to 79 wherein G is C 1 -C 6 -alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26
81. A pharmaceutical composition according to any one of the claims 60 or 78 to 79 wherein G is Cl-Co-alkyl or ArG1, wherein the aryl is optionally substituted with up to three substitu 15 ents R 24 , R 25 and R 2 6 .
82. A pharmaceutical composition according to claim 80 wherein G is Cl-C 6 -alkyl.
83. A pharmaceutical composition according to claim 82 wherein G is phenyl optionally sub stituted with up to three substituents R 24 , R 25 and R 26
84. A pharmaceutical composition according to any one of the claims 60 to 83 wherein R 24 20 R 2s and R 26 are independently selected from *hydrogen, halogen, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -SCF 3 , NO 2 , -OR 27 , -NR 27 R 28 , -SR 27 , -C(O)NR 27 R 28 , -OC(O)NR 27 R 28 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -CH 2 C(O)NR 27 R 28 , -OCH 2 C(O)NR 27 R 28 , -CH 2 0R 27 , -CH 2 NR 2 7 R 28 , 25 -OC(O)R 2 7 , -OCl-C 6 -alkyl-C(O)OR 27 , -SC 1 -C 6 -alkyl-C(O)OR 2 7 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -NR 27 -C(=O)-C l -C 6 -alkyl-C(=O) O R 27 , -NR 27 -C(=O)-C 1 -C 6 alkenyl-C(=O)OR-, -C(=O)NR 27 -C 1 -C 6 -alkyl-C(=O)OR 7 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 7 , 30 * Cl-Ce-alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 29 WO 2004/080480 PCT/DK2004/000158 434 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-C 1 -C 6 -alkyl, heteroaryl-C 2 -C6 alkenyl or heteroaryl-C 2 -C 6 -alkynyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 .
85. A pharmaceutical composition according to claim 84 wherein R 2 4 , R 2, 5 and R 26 are inde pendently selected from 10 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 2 8 , -NR 27 C(O)OR 2 8 , -OC(O)R 27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC 1 -C 6 -alkyl-C(O)OR 2 7 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)N R 27 -Cl 1 -C 6 -alkyl-C(=O)OR 2 7 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 27 , 15 * Cl-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents independently se lected from R 20 *aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cs-C 6 -alkyl, heteroaryl-C 2 -C 6 alkenyl or heteroaryl-C 2 -C 6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu 25 ents selected from R3.
86. A pharmaceutical composition according to claim 85 wherein R 24 , R 25 and R 26 are inde pendently selected from * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , 30 -OC(O)R 27 , -OC,-C 6 -alkyl-C(O)OR 27 , -SC 1 -C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)NR 27 -Cl -C 6 -alkyl-C(=O)OR 2 7 , -C1-C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 27 * Cl-C 6 -alkyl optionally substituted with one or more substituents independently se 35 lected from R 29 WO 2004/080480 PCT/DK2004/000158 435 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30 .
87. A pharmaceutical composition according to claim 86 wherein R 21 , R 22 and R 23 are inde pendently selected from 10 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 2 8 -OC(O)R 27 , -OC 1 -C 6 -alkyl-C(O)OR 27 , -SC,-C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=O)OR 27 , -C(=O)NR 27 -C -C 6 -alkyl-C(=O)OR 27 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 7 , 15 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 29 * ArG1, ArG1-O-, ArG1-C(O)-, ArGl1-Cl-C 6 -alkoxy, ArGl-C 1 -CB-alkyl, Het3, Het3-C 1 C 6 -alkyl 20 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 30
88. A pharmaceutical composition according to claim 87 wherein R 21 , R 22 and R 23 are inde pendently selected from 25 * hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 2 8 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OC 1 -C 6 -alkyl-C(O)OR 27 , -SC 1 -C 6 -alkyl-C(O)OR 27 , -C 2 -C 6 -alkenyl C(=0)OR 27 , -C(=O)NR 27 -Cl-C 6 -alkyl-C(=O)OR 27 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 7 , 30 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R' *ArG1, ArG1-O-, ArGI-C(O)-, ArGl-Cl-C 6 -alkoxy, ArGl-Cl-CB-alkyl, Het3, Het3-Cl C 6 -alkyl WO 2004/080480 PCT/DK2004/000158 436 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 30
89. A pharmaceutical composition according to claim 88 wherein R 21 , R 22 and R 23 are inde pendently selected from 5 *hydrogen, halogen, -OCF 3 , -OR 27 , -NR 27 R 28 , -SR 27 , -NR 27 C(O)R 28 , -NR 27 C(O)OR 28 , -OC(O)R 27 , -OCl-C 6 -alkyl-C(O)OR 27 , -SCI-C 6 -alkyl-C(O)OR 27 , -C 2 -Ce-alkenyl C(=O)OR 27 , -C(=O)N R 27 -Cl -C 6 -alkyl-C(=O)OR 27 , -C 1 -C 6 -alkyl-C(=O)OR 27 , or -C(O)OR 2 7 , 10 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 9 *ArG1, ArG1-O-, ArG1-Cl-C 6 -alkoxy, ArGl-Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se 15 lected from R.
90. A pharmaceutical composition according to any one of the claims 60 or 78 to 89 wherein R 2 0 is hydrogen or methyl.
91. A pharmaceutical composition according to claim 90 wherein R 2 is hydrogen.
92. A pharmaceutical composition according to any one of the claims 60 or 78 to 91 wherein 20 R 27 is hydrogen, CI-C 6 -alkyl or aryl.
93. A pharmaceutical composition according to claim 92 wherein R 27 is hydrogen or Cj-C 6 alkyl or ArG1.
94. A pharmaceutical composition according to claim 93 wherein R 2 7 is hydrogen or Cl-C 6 alkyl. 25 95. A pharmaceutical composition according to any one of the claims 60 or 78 to 93 wherein R 28 is hydrogen or Cl-CB-alkyl.
96. A pharmaceutical composition according to claim 60 wherein R 17 and R 18 are independ ently selected from 30 *hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 27 , -NR 27 R 28 , -SR 27, -S(O)R 27 -S(O) 2 R 27 , -C(O)NR 27 R 28 , -CH 2 0R 2 7 , -OC(O)R 27 , -OCl-C 6 -alkyl-C(O)OR 2 7 , -SC 1 -C 6 alkyl-C(O)OR 27 , or-C(O)OR 7 , * C,-C6-alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, optionally substituted with one or more 35 substituents independently selected from R 29 WO 2004/080480 PCT/DK2004/000158 437 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl, 5 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 .
97. A pharmaceutical composition according to claim 96 wherein R 1 7 and R 18 are independ ently selected from 10 e hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 2 7 R 28 , or -C(O)OR 27 , * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se lected from R2 15 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-C 1 -Ce-alkyl, heteroaryl, heteroaryl-Cl-C 6 alkyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 . 20 98. A pharmaceutical composition according to claim 97 wherein R 1 7 and R 1 8 are independ ently selected from * hydrogen, halogen, -CN, -CF3, -NO 2 , -OR 27 , -NR 27 R 2 8 , or -C(O)OR 2 7 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R 9 25 *aryl, aryloxy, aroyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, heteroaryl, heteroaryl-C- 1 -C 6 alkyl of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 3 .
99. A pharmaceutical composition according to claim 98 wherein R 17 and R 1 " are independ 30 ently selected from *hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 , -NR 27 R 2 8 , or -C(O)OR 27 * methyl, ethyl propyl optionally substituted with one or more substituents independ ently selected from R *ArG1, ArG1-O-, ArG1-C(O)-, ArG1-C 1 -C 6 -alkoxy, ArGl-Cl-C 6 -alkyl, Het3, Het3-Cl 35 C 6 -alkyl WO 2004/080480 PCT/DK2004/000158 438 of which the cyclic moieties optionally may be substituted with one or more substituents se lected from R 3 .
100. A pharmaceutical composition according to claim 99 wherein R 1 7 and R 1 8 are independ ently selected from 5 * hydrogen, halogen, -CN, -CF3, -NO 2 , -OR 27 , -NR 2 7 R 2 8 , or-C(O)OR 27 * C,-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 2 9 * phenyl, phenyloxy, phenyl-Cl-C 6 -alkoxy, phenyl-Cl-C 6 -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se 10 lected from R 3 .
101. A pharmaceutical composition according to any one of the claims 60 to 100 wherein R 2 7 is hydrogen or CI-C 6 -alkyl.
102. A pharmaceutical composition according to claim 101 wherein R 27 is hydrogen, methyl or ethyl. 15 103. A pharmaceutical composition according to any one of the claims 60 to 102 wherein R 28 is hydrogen or C 1 -C6-alkyl.
104. A pharmaceutical composition according to claim 103 wherein R 28 is hydrogen, methyl or ethyl.
105. A pharmaceutical composition according to any one of the claims 60 to 104 wherein R 72 20 is -OH or phenyl.
106. A pharmaceutical composition according to claim 60 wherein CGr is N H
107. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is of the form H-I-J 25 wherein H is 0 OH O OH HO O OHO / HO or HO or N H WO 2004/080480 PCT/DK2004/000158 439 wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 I is selected from 5 * a valence bond, * -CH 2 N(R 32 )- or -SO 2 N(R 33 )-, z 2 * wherein Z 1 is S(O) 2 or CH 2 , Z 2 is -NH-, -O-or -S-, and n is 1 or 2, J is 10 * C 1 -C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R 34, * Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-Cl-C 6 -alkoxy-, aryl-C 1 -C 6 -alkyl-, aryl-C2 C 6 -alkenyl-, aryl-C 2 -C 6 -alkynyl-, heteroaryl, heteroaryl-Cl-C 6 -alkyl-, heteroaryl-C 2 -C 6 alkenyl- or heteroaryl-C 2 -C 6 -alkynyl-, wherein the cyclic moieties are optionally substi 15 tuted with one or more substituents selected from R 37 , * hydrogen, R 3 1 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 35 , -C(0)R 3 5 , -NRR35 36 , -SR 35 , 20 -NR 35 S(0) 2 R 36 , -S(0) 2 NR3R 36 , -S(O)NR 35 R 36 , -S(O)R 3 5 , -S(O) 2 R 35 , -C(O)NR 3 5 R 36 , -OC(O)NR 35 R 36 , -NR 35 C(0)R 36 , -CH 2 C(0)NR 35 R 3 6 , -OCH 2 C(O)NR 35 R 36 , -CH 2 0R 3 5 , -CH 2 NR 35 R 36 , -OC(0)R 35 , -OC,-C 6 -alkyl-C(O)OR 3 5 , -SCl-C 6 -alkyl-C(O)OR 3 5 -C 2 -C 6 -alkenyl C(=O)OR 35 , -NR 35 -C(=0)-C-C-alkyl-C(=0)OR35, -NR 35 -C(=O)-Cl-C 6 -alkenyl-C(=O)OR3, Cl-C 6 -alkyl, CI-C 6 -alkanoyl or -C(O)OR 5 , ?5 R 32 and R 33 are independently selected from hydrogen, Cl-C 6 -alkyl or C 1 -C 6 -alkanoyl, R3 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR , and -NR 35 R 36 , !0 R 3 5 and R 36 are independently selected from hydrogen, C 1 -C 6 -alkyl, aryl-Cl-C 6 -alkyl or aryl, or R 35 s and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- WO 2004/080480 PCT/DK2004/000158 440 eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 37 is independently selected from halogen, -C(O)OR 35 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , 5 OR" , -NR 35 R 36 , CI-C 6 -alkyl or Cl-C 6 -alkanoyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
108. A pharmaceutical composition according to claim 107 wherein CGr is of the form H-I-J, 10 wherein H is 0 0 0 OH HO HO HO O HO or O N H wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 , I is selected from 15 *a valence bond, * -CH 2 N(R 32 )- or -SO 2 N(R 33 )-, -Z-N- i n * wherein Z 1 is S(O) 2 or CH 2 , Z 2 is N,-O-or -S-, and n is 1 or 2, J is 20 * Cl-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R3, *Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-Cl-C 6 -alkoxy-, aryl-Cl-C 6 -alkyl-, aryl-C 2 C 6 -alkenyl-, aryl-C 2 -C 6 -alkynyl-, heteroaryl, heteroaryl-Cl-C 6 -alkyl-, heteroaryl-C 2 -C 6 alkenyl- or heteroaryl-C 2 -C 6 -alkynyl-, wherein the cyclic moieties are optionally substi 25 tuted with one or more substituents selected from R 37 * hydrogen, WO 2004/080480 PCT/DK2004/000158 441 R 3 1 ' is independently selected from hydrogen, halogen, -CN, -CHzCN, -CHF 2 , -CF 3 , -OCFa, -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 35 , -C(O)R 35 , -NR 35 R 3 , -SR, -NR 35 S(O) 2 R 36 , -S(O) 2 NR 35 R 3 6 , -S(O)NR 3 5 sR 36 , -S(O)R 35 , -S(O) 2 R 35 , -C(O)NR 35 R 36 , -OC(O)NR 3 5 R 36 , -NR"C(O)R 3 6 , -CH 2 C(O)NR"R 36 , -OCH 2 C(O)NR 35 R 3 6 , -CH 2 0R 35 , 5 -CH 2 NR 35 R 36 , -OC(O)R 3 5 , -OC,-C 6 -alkyl-C(O)OR", -SC 1 -Co-alkyl-C(O)OR 35 -C 2 -C 6 -alkenyl C(=O)OR 35 , -NR3-C(=O)-Cl-C 6 -alkyl-C(=O)OR", -NR 3 5 -C(=O)-Cl-C-alkenyl-C(=O)OR 35 _ C-C 6 -alkyl, C 1 -C 6 -alkanoyl or -C(O)ORa, R 32 and R 33 are independently selected from hydrogen, Cl-C 6 -alkyl or C 1 -C 6 -alkanoyl, 10 R 34 is independently selected from halogen, -CN, -CF3, -OCF 3 , -OR , and -NR 35 R 3 , R 35 s and R 36 are independently selected from hydrogen, C,-C 6 -alkyl, aryl-Cl-C 6 -alkyl or aryl, or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom 15 may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 37 is independently selected from halogen, -C(O)OR 3 5 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , 20 OR 3 , -NR 35 R 36 , C 1 -C 6 -alkyl or C 1 -C 6 -alkanoyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R 31 and J cannot both be hydrogen. 25 109. A pharmaceutical composition according to any one of the claims 107 or 108 wherein H is O O HO HO or HO HO
110. A pharmaceutical composition according to claim 109 wherein H is WO 2004/080480 PCT/DK2004/000158 442 0 HO HO
111. A pharmaceutical composition according to claim 109 wherein H is 0 HO HO
112. A pharmaceutical composition according to any one of the claims 107 to 111wherein I is 5 a valence bond, -CH 2 N(R 32 )-, or-SO 2 N(R 33 )-.
113. A pharmaceutical composition according to claim 112 wherein I is a valence bond.
114. A pharmaceutical composition according to any one of the claims 107 to 113 wherein J is * hydrogen, 10 * Cl-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 3 5 , and -NR 3 sR 3 6 , *aryl, or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 3 . 15 115. A pharmaceutical composition according to claim 114 wherein J is * hydrogen, * aryl or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 3 .
116. A pharmaceutical composition according to claim 114 wherein J is 20 *hydrogen, eArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 3 .
117. A pharmaceutical composition according to claim 116 wherein J is * hydrogen, 25 * phenyl or naphthyl optionally substituted with one or more substituents inde pendently selected from R 37
118. A pharmaceutical composition according to claim 117 wherein J is hydrogen. WO 2004/080480 PCT/DK2004/000158 443
119. A pharmaceutical composition according to any one of the claims 107 to 118 wherein R 32 and R 33 are independently selected from hydrogen or Cl-C 6 -alkyl.
120. A pharmaceutical composition according to any one of the claims 107 to 119 wherein R3 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -ORa, -C(O)R 35 , -NR 35 R 3 , -SR 3 , 5 -C(O)NRMR 36 , -OC(O)NR"R 36 , -NRaC(O)R 3 6 , -OC(O)Ra, -OCI-C 6 -alkyl-C(O)OR 35 , -SCI-C 6 alkyl-C(O)OR s or -C(O)OR 5 .
121. A pharmaceutical composition according to claim 120 wherein R 34 is hydrogen, halogen, -CF 3 , -NO 2 , -OR 3 5 , -NR 3 5 R 3 6 , -SR 3 5 , -NR 35 C(O)R 3 6 , or -C(O)OR".
122. A pharmaceutical composition according to claim 121 wherein R3 is hydrogen, halogen, 10 -CF 3 , -NO 2 , -OR", -NR 35 R 3 6 , or -NR 3 5 C(O)R 36 .
123. A pharmaceutical composition according to claim 122 wherein R3 is hydrogen, halogen, or -OR 35 .
124. A pharmaceutical composition according to any one of the claims 107 to 123 wherein R3 and R3 are independently selected from hydrogen, C 1 -C6-alkyl, or aryl. 15 125. A pharmaceutical composition according to claim 124 wherein R3 and R 36 are inde pendently selected from hydrogen or Cl-C 6 -alkyl.
126. A pharmaceutical composition according to any one of the claims 107 to 125 wherein R 37 is halogen, -C(O)OR 35 , -CN, -CF3, -OR 35 , -NR 35 R 36 , C 1 -C 6 -alkyl or CI-C 6 -alkanoyl.
127. A pharmaceutical composition according to claim 126 wherein R 37 is halogen, 20 C(O)OR 3 5 , -OR 35 , -NR 35 R 36 , Cl-C 6 -alkyl or C 1 -C 6 -alkanoyl.
128. A pharmaceutical composition according to claim 127 wherein R 37 is halogen, C(O)OR or -OR 3 .
129. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is HN N K\ Q \ M N=N T 25 wherein K is a valence bond, Cl-C 6 -alkylene, -NH-C(=O)-U-, -CI-C 6 -alkyl-S-, -Cl-Cs-alkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any C 1 -C 6 -alkyl moiety is optionally substituted with R3, U is a valence bond, C 1 -C 6 -alkenylene, -Cl-C 6 -alkyl-O- or Cl-C 6 -alkylene wherein any Cj 30 C 6 -alkyl moiety is optionally substituted with C 1 -C 6 -alkyl, R3 is CI-C 6 -alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R 39 , WO 2004/080480 PCT/DK2004/000158 444 R 39 is independently selected from halogen, cyano, nitro, amino, M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are 5 optionally substituted with one or more substituents independently selected from R 40 , R 4 is selected from *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , .-OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -OS(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 41 , -NR41R 42 , -SR 41 , 10 -NR 41 S(O) 2 R 42 , -S(O) 2 NR 41 R 42 , -S(O)NR 41 R 42 , -S(O)R 41 , -S(O) 2 R 41 , -OS(O) 2 R 4 1 , -C(O)NR 41 R 42 , -OC(O)NR 41 R 42 , -NR 41 C(O)R 42 , -CH 2 C(O)NR 4 1 R 42 , -OC 1 -C 6 alkyl-C(O)NR 41 R 42 , -CH 2 0R 4 1 , -CH 2 OC(O)R 41 , -CH 2 NR 41 R 42 , -OC(O)R 41 , -OC1-C6 alkyl-C(O)OR 4 1 , -OC 1 -C 6 -alkyl-OR 41 , -S-Cj-C 6 -alkyl-C(O)OR 41 , -C 2 -C6-alkenyl C(=O)OR 41 , -NR 4 1 -C(=O)-C 1 -C 6 -alkyl-C(=O)OR 41 , -NR 4 1 -C(=O)-C,-C 6 15 alkenyl-C(=O)OR 4 1 , -C(O)OR 41 , -C 2 -C 6 -alkenyl-C(=O)R 41 , =0, -NH-C(=O)-O-Cl C 6 -alkyl, or -NH-C(=O)-C(=O)-O-C 1 -C 6 -alkyl, * C 1 -C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -C 6 -alkynyl, which may each optionally be substituted with one or more substituents selected from R 43 , 20 *aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C 1 -C 6 -alkoxy, aryl-Cl-C6-alkyl, aryl-C 2 -C 6 -alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C6-alkynyl, heteroaryl, heteroaryl-Cl C 6 -alkyl, heteroaryl-C 2 -C 6 -alkenyl or heteroary-C 2 -C 6 -alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from 25 R , R 4 1 and R 42 are independently selected from hydrogen, -OH, C 1 -C 6 -alkyl, Cl-C 6 -alkenyl, aryl C 1 -Ce-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substi 30 tuted with one or more substituents independently selected from R 46 ; R 41 and R 42 when at tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 35 WO 2004/080480 PCT/DK2004/000158 445 R 43 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 4 1 , and -NR 41 R 42 R" is independently selected from halogen, -C(O)OR 41 , -CH 2 C(O)OR 41 , -CH 2 0R 41 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR 41 , -NR 41 R 42 and C 1 -C 6 -alkyl, 5 R is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -O-C 1 -C 6 -alkyl, -C(O)-O-Cl C 6 -alkyl, -COOH and -NH 2 , R 46 is independently selected from halogen, -C(O)OC-C-alkyl, -COOH, -CN, -CF 3 , -OCF 3 , NO 2 , -OH, -OC,-Ce-alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, 10 Q is a valence bond, Cl-C 6 -alkylene, -C-Co 6 -alkyl-O-, -Cl-C 6 -alkyl-NH-, -NH-Cj-C 6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-Cl-C 6 -alkyl, -C(=O)-, or -Cl-C 6 -alkyl-C(=O)-N(R 4 7 )- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R4, 15 R 47 and R 48 are independently selected from hydrogen, C 1 -Co-alkyl, aryl optionally substituted with one or more R 49 R 49 is independently selected from halogen and -COOH, 20 T is * hydrogen, * C,-C 6 -alkyl, C 2 -Ce-alkenyl , C 2 -C6-alkynyl, C 1 -C 6 -alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents 25 independently selected from R 5 0 , * aryl, aryloxy, aryloxy-carbonyl, aryl-Cs-C 6 -alkyl, aroyl, aryl-Cl-C 6 -alkoxy, aryl-C2 C 6 -alkenyl, aryl-C 2 -C 6 -alkyny-, heteroaryl, heteroaryl-Cl-C 6 -alkyl, heteroaryl-C2 C 6 -alkenyl, heteroaryl-C 2 -C6-alkynyl, 30 wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 50 , R s 50 is Cl-C 6 -alkyl, Cl-Co-alkoxy, aryl, aryloxy, aryl-C,-C 6 -alkoxy, -C(=O)-NH-Cj-C 6 -alkyl-aryl, -C(=O)-NRSOA_ -C-alkyl, -C(=O)-NH-(CH 2 CH 2 0)mC,-C-alkyl-COOH, heteroaryl, het 35 eroaryl-Cs-C 6 -alkoxy, -Cl-C 6 -alkyl-COOH, -O-C-C 6 -alkyl-COOH, -S(O) 2 R' 1 , WO 2004/080480 PCT/DK2004/000158 446 -C 2 -C 6 -alkenyl-COOH, -OR 5 1 , -NO 2 , halogen, -COOH, -CF 3 , -CN, =0, -N(R 5 1 R 52 ), wherein m is 1, 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R 53 , and the alkyl moieties are optionally substituted with one or more RoB. R 5 0A and Rs 5 B are independently selected from -C(O)OC-C 6 -alkyl, -COOH, -Cl-C 6 -alkyl 5 C(O)OC-C 6 -alkyl, -C-C 6 -alkyl-COOH, or Cl-C 6 -alkyl, R 5 1 and R 52 are independently selected from hydrogen and Cl-C 6 -alkyl, R 53 is independently selected from Cl-C 6 -alkyl, CI-C 6 -alkoxy, -Cl-C 6 -alkyl-COOH, -C2 C 6 -alkenyl-COOH, -OR 5 1 , -NO 2 , halogen, -COOH, -CF 3 , -CN, or -N(R 5 ,R 5 2 ), 10 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
130. A pharmaceutical composition according to claim 129 wherein K is a valence bond, C CB-alkylene, -NH-C(=O)-U-, -C-C 6 -alkyl-S-, -C1-C 6 -alkyl-O-, or -C(=O)-, wherein any C 1 C 6 -alkyl moiety is optionally substituted with R3. 15 131. A pharmaceutical composition according to claim 130 wherein K is a valence bond, C 1 C-alkylene, -NH-C(=O)-U-, -C-C 6 -alkyl-S-, or -C-C 6 -alkyl-O, wherein any Cl-C 6 -alkyl moiety is optionally substituted with R 38 .
132. A pharmaceutical composition according to claim 131 wherein K is a valence bond, C C-alkylene, or -NH-C(=O)-U, wherein any Cl-C 6 -alkyl moiety is optionally substituted with 20 R 38 .
133. A pharmaceutical composition according to claim 132 wherein K is a valence bond or C0 1 -C 6 -alkylene, wherein any Cl-C 6 -alkyl moiety is optionally substituted with R 38 .
134. A pharmaceutical composition according to claim 132 wherein K is a valence bond or -NH-C(=O)-U. 25 135. A pharmaceutical composition according to claim 133 wherein K is a valence bond.
136. A pharmaceutical composition according to any one of the claims 129 to 135 wherein U is a valence bond or -Cl-C 6 -alkyl-O-.
137. A pharmaceutical composition according to claim 136 wherein U is a valence bond.
138. A pharmaceutical composition according to any one of the claims 129 to 137 wherein M 30 is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 4 .
139. A pharmaceutical composition according to claim 138 wherein M is ArG1 or Hetl, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 WO 2004/080480 PCT/DK2004/000158 447
140. A pharmaceutical composition according to claim 139 wherein M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40
141. A pharmaceutical composition according to claim 140 wherein M is ArG1 or Het3, 5 wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40
142. A pharmaceutical composition according to claim 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from R 40
143. A pharmaceutical composition according to claim 141 wherein M is indolylene optionally 10 substituted with one or more substituents independently selected from R 40 .
144. A pharmaceutical composition according to claim 143 wherein M is R40
145. A pharmaceutical composition according to claim 141 wherein M is carbazolylene op tionally substituted with one or more substituents independently selected from R 40 . 15 146. A pharmaceutical composition according to claim 145 wherein M is R 4 0
147. A pharmaceutical composition according to any one of the claims 129 to 146 wherein R 40 is selected from hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 4 1 , -NR 41 R 42 , -SR 41 , -S(O) 2 R 41 20 -NR 4 1 C(O)R 42 , -OCl-C 6 -alkyl-C(O)NR 41R 4 2 , -C 2 -C 6 -alkenyl-C(=O)OR 41 , -C(O)OR 41 , =O, -NH-C(=O)-O-C-C 6 -alkyl, or -NH-C(=O)-C(=O)-O-CI-C 6 -alkyl, CI-C 0 -alkyl or C 2 -C 6 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 25 * aryl, aryloxy, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, aryl-C 2 -C 6 -alkenyl, heteroaryl, het eroaryl-Cl-C 6 -alkyl, or heteroaryl-C 2 -C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R".
148. A pharmaceutical composition according to claim 147 wherein R 4 0 is selected from WO 2004/080480 PCT/DK2004/000158 448 *hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 41 , -NR 41 R 42 , -SR 41 , -S(O) 2 R 4 1 , -NR 41 C(O)R 42 , -OCI-C 6 -alkyl-C(O)NR 4 1 R 42 , -C 2 -C 6 -alkenyl-C(=O)OR 41 , -C(O) O R 4 1 , =O, -NH-C(=O)-O-C-C 6 -alkyl, or -NH-C(=O)-C(=O)-O-C 1 -C 6 -alkyl, 5 Cl-C 6 -alkyl or C 2 -CB 5 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 , *ArG1, ArG1-O-, ArGl-Ci-C 6 -alkoxy, ArG1-C 1 -C 6 -alkyl, ArG1-C 2 -C 6 -alkenyl, Het3, Het3-C 1 -C 6 -alkyl, or Het3-C 2 -C 6 -alkenyl, wherein the cyclic moieties optionally may be 10 substituted with one or more substituents selected from R".
149. A pharmaceutical composition according to claim 148 wherein R 4 is selected from e hydrogen, halogen, -CF 3 , -NO 2 , -OR 41 , -NR 41 R 42 , -C(O)OR 41 , =0, or-NR 41 C(O)R 42 , * C 1 -C 6 -alkyl, * ArG 1. 15 150. A pharmaceutical composition according to claim 149 wherein R 4 0 is hydrogen.
151. A pharmaceutical composition according to claim 149 wherein R 40 is selected from * halogen, -NO 2 , -OR 41 , -NR 41 R 4 2 , -C(O)OR 41 , or -NR 41 C(O)R 42 , * methyl, * phenyl. 20 152. A pharmaceutical composition according to any one of the claims 129 to 151 wherein R 41 and R 42 are independently selected from hydrogen, C 1 -C 6 -alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
153. A pharmaceutical composition according to claim 152 wherein R 41 and R 42 are inde pendently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may 25 optionally be substituted with halogen or-COOH.
154. A pharmaceutical composition according to any one of the claims 129 to 153 wherein Q is a valence bond, Cl-C 6 -alkylene, -Cl-C 6 -alkyl-O-, -C 1 -C 6 -alkyl-NH-, -NH-Cl-C 6 -alkyl, -NH-C(=O)-, -C(=O)-NH-, -O-C 1 -C 6 -alkyl, -C(=O)-, or -C 1 -C 6 -alkyl-C(=O)-N(R 47 )- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected 30 from R4.
155. A pharmaceutical composition according to claim 154 wherein Q is a valence bond, -CH 2 -, -CH 2 -CH 2 -, -CH 2 -O-, -CH 2 -CH 2 -O-, -CH 2 -NH-, -CH 2 -CH 2 -NH-, -NH-CH 2 -, -NH-CH 2 -CH 2 -, -NH-C(=O)-, -C(=O)-NH-, -O-CH 2 -, -O-CH 2 -CH 2 -, or -C(=O)-.
156. A pharmaceutical composition according to any one of the claims 129 to 155 wherein 35 R 47 and R 48 are independently selected from hydrogen, methyl and phenyl. WO 2004/080480 PCT/DK2004/000158 449
157. A pharmaceutical composition according to any one of the claims 129 to 156 wherein T is * hydrogen, * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se 5 lected from R 50, earyl, aryl-Cl-C 6 -alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 50 .
158. A pharmaceutical composition according to claim 157 wherein T is * hydrogen, 10 * Cl-C 6 -alkyl optionally substituted with one or more substituents independently se lected from R5, *ArG1, ArG1-Cl-CB-alkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are op tionally substituted with one or more substituents independently selected from R 5 .
159. A pharmaceutical composition according to claim 158 wherein T is 15 * hydrogen, * C 1 -C 6 -alkyl, optionally substituted with one or more substituents independently se lected from R 5 0, o phenyl, phenyl-Cl-C 6 -alkyl, wherein the alkyl and phenyl moieties are optionally substituted with one or more substituents independently selected from R 5 0 . 20 160. A pharmaceutical composition according to claim 159 wherein T is phenyl substituted with R 5 0 .
161. A pharmaceutical composition according to any one of the claims 129 to 160 wherein R5 is C 1 l-C 6 -alkyl, Cl-C6-alkoxy, aryl, aryloxy, aryl-Cl-C 6 -alkoxy, -C(=O)-NH-C 1 -C 6 -alkyl-aryl, -C(=O)-NR50A-C -C 6 -alkyl, -C(=O)-NH-(CH 2 CH20)mCI-C6-alkyl-COOH, heteroaryl, -Cl 25 C 6 -alkyl-COOH, -O-C 1 -C 6 -alkyl-COOH, -S(O) 2 R 5 ', -C 2 -C 6 -alkenyl-COOH, -OR - ', -NO 2 , halo gen, -COOH, -CF3, -CN, =0, -N(R 5 s 1 R 5 2 ), wherein the aryl or heteroaryl moieties are option ally substituted with one or more R 5 .
162. A pharmaceutical composition according to claim 161 wherein R s 50 is CI-C 6 -alkyl, Cj C 6 -alkoxy, aryl, aryloxy, -C(=O)-NR50A-C1-C6-alkyl, -C(=O)-NH-(CH 2 CH 2 0)mCl-C 6 -alkyl 30 COOH, aryl-Cl-C 6 -alkoxy , -OR s 51 , -NO 2 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
163. A pharmaceutical composition according to claim 162 wherein R 50 sis Cl-C 6 -alkyl, ary loxy, -C(=O)-NRSAC -C 6 -alkyl, -C(=O)-NH-(CH 2 CH 2 0)mCl-C 6 -alkyl-COOH, aryl-Cl C 6 -alkoxy, -OR 5 ', halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted 35 with one or more R 53 . WO 2004/080480 PCT/DK2004/000158 450
164. A pharmaceutical composition according to claim 163 wherein R" is Cl-C 6 -alkyl, ArGI1-O-, -C(=O)-NR50A-C-C6-alkyl , -C(=O)-NH-(CH 2 CH 2 0)mCI-C 6 -alkyl-COOH, ArGI1-Cj C 6 -alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R s 53 . 5 165. A pharmaceutical composition according to claim 164 wherein R6 0 is -C(=O)-NR50ACH 2 , -C(=O)-NH-(CH 2 CH 2 0) 2 CH 2 1-COOH, or -C(=O)-NR50ACH 2 CH 2 .
166. A pharmaceutical composition according to claim 164 wherein R ° is phenyl, methyl or ethyl.
167. A pharmaceutical composition according to claim 166 wherein R" is methyl or ethyl. 10 168. A pharmaceutical composition according to any one of the claims 129 to 167 wherein m is 1 or 2.
169. A pharmaceutical composition according to any one of the claims 129 to 168 wherein R 51 is methyl.
170. A pharmaceutical composition according to any one of the claims 129 to 169 wherein 15 R 53 is CI-Cs-alkyl, CI-C 6 -alkoxy, -OR 5 1 , halogen,or -CF 3 .
171. A pharmaceutical composition according to any one of the claims 129 to 170 wherein R 5 0A is -C(O)OCH 3 , -C(O)OCH 2 CH 3 -COOH, -CH 2 C(O)OCH 3 , -CH 2 C(O)OCH 2 CH 3 , -CH 2 CH 2 C(O)OCH 3 , -CH 2 CH 2 C(O)OCH 2 CH 3 , -CH 2 COOH, methyl, or ethyl.
172. A pharmaceutical composition according to any one of the claims 129 to 171 wherein 20 R s 50B is -C(O)OCH 3 , -C(O)OCH 2 CH 3 -COOH, -CH 2 C(O)OCH 3 , -CH 2 C(O)OCH 2 CH 3 , -CH 2 CH 2 C(O)OCH 3 , -CH 2 CH 2 C(O)OCH 2 CH 3 , -CH 2 COOH, methyl, or ethyl.
173. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is ,N --N HN N S 25 wherein V is C 1 -C 6 -alkyl, aryl, heteroaryl, aryl-C 1 l_-alkyl- or aryl-C 2 -6-alkenyl-, wherein the al kyl or alkenyl is optionally substituted with one or more substituents independently selected from R " , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 55 , 30 R " is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 , R 55 s is independently selected from WO 2004/080480 PCT/DK2004/000158 451 *hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF3, -OCF 3 , -OCHF 2 , -OCH 2 CFa, -OCF 2 CHF 2 , -S(O) 2 CF 3 , -OS(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 56 , -NR6R 57 , -SR 5 , -NR 56 S(O) 2 R 5 7 , -S(O) 2 NR 56 R 5 7 , -S(O)NRR 57 , -S(O)R 56, -S(O) 2 R 5 6 , -OS(O) 2 R 56 , -C(O)NR 6 R 57 , -OC(O)NR" 6 R 5 7 , -NR 56 C(O)R 57 , -CH 2 C(O)NR 56 R 57 , -OC 1 -C 6 5 alkyl-C(O)NRR 57 , -CH 2 OR 56, -CH 2 OC(O)R e 56 , -CH 2 NR 56 R 5 7 , -OC(O)R", -OC-Cs alkyl-C(O)OR 5 6, -OC 1 l-C 6 -alkyl-OR 6, -SCI-C 6 -alkyl-C(O)OR 5 6 , -C 2 -C 6 -alkenyl C(=O)OR 56 , -NR 56 -C(=O)-C 1 -C 6 -alkyl-C(=O)OR 5 6 , -NR"-C(=O)-Cj-C 6 alkenyl-C(=O)OR , -C(O)OR - ", or -C 2 -C6-alkenyl-C(=O)R, 10 * Cl-C 6 -alkyl, Cz-C 6 -alkenyl or C 2 -C 6 -alkynyl, which may optionally be substituted with one or more substituents selected from R 8 , *aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Cl-C 6 -alkoxy, aryl-Cl-C 6 -alkyl, 15 aryl-C 2 -C 6 -alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-Cl C 6 -alkyl, heteroaryl-C 2 -C 6 -alkenyl or heteroaryl-C 2 -C 6 -alkynyl, of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 59 , 20 R56 and R 57 are independently selected from hydrogen, OH, CF3, C 1 -C 1 2 -alkyl, aryl-Cl-C 6 alkyl, -C(=O)-Cl-C 6 -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 60 , and the aryl groups may option ally be substituted with one or more substituents independently selected from R 61 ; R 5 6 and 25 R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, 30 R 58 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR s 56 , and -NRR s 57 , R 59 is independently selected from halogen, -C(O)OR", -CH 2 C(O)OR s6 , -CH 2 OR5, -CN, CF3, -OCF 3 , -NO 2 , -OR", -NR"R 57 and Cl-C 6 -alkyl, WO 2004/080480 PCT/DK2004/000158 452 R'o is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OC-C 6 -alkyl, -C(O)OC-C 6 alkyl, -C(=O)-R 62 , -COOH and -NH 2 , R 61 is independently selected from halogen, -C(O)OC-C 6 -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , 5 NO 2 , -OH, -OC,-C-alkyl, -NH 2 , C(=O) or C,-C 6 -alkyl, R 62 is Cl-C 6 -alkyl, aryl optionally substituted with one or more substituents independently se lected from halogen, or heteroaryl optionally substituted with one or more C 1 -C 6 -alkyl inde pendently, 10 or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
174. A pharmaceutical composition according to claim 173 wherein V is aryl, heteroaryl, or aryl-Cl_6-alkyl-, wherein the alkyl is optionally substituted with one or more substituents inde pendently selected R5, and the aryl or heteroaryl is optionally substituted with one or more 15 substituents independently selected from R 55 .
175. A pharmaceutical composition according to claim 174 wherein V is aryl, Hetl, or aryl-C. 6 -alkyl-, wherein the alkyl is optionally substituted with one or more substituents independ ently selected from R " , and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 . 20 176. A pharmaceutical composition according to claim 175 wherein V is aryl, Het2, or aryl-C_ 6 -alkyl-, wherein the alkyl is optionally substituted with one or more substituents independ ently selected from R " , and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 .
177. A pharmaceutical composition according to claim 176 wherein V is aryl, Het3, or aryl-C. 25 6 -alkyl-, wherein the alkyl is optionally substituted with one or more substituents independ ently selected from R5, and the aryl or heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 55 .
178. A pharmaceutical composition according to claim 177 wherein V is aryl optionally substi tuted with one or more substituents independently selected from R 55 . 30 179. A pharmaceutical composition according to claim 178 wherein V is ArG1 optionally sub stituted with one or more substituents independently selected from R 5 .
180. A pharmaceutical composition according to claim 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independently selected from R 5 5 . WO 2004/080480 PCT/DK2004/000158 453
181. A pharmaceutical composition according to claim 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R 55 .
182. A pharmaceutical composition according to any one of the claims 173 to 181 wherein R 55 is independently selected from 5 halogen, C 1 -C 6 -alkyl, -CN, -OCF 3 ,-CF 3 , -NO 2 , -OR", -NR 5 6 R s 5 7 , -NR"C(O)R 5 7 -SR 56 , -OC-C 8 -alkyl-C(O)OR 5 6 , or -C(O)OR", * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se lected from R s8 * aryl, aryl-Cl-C 6 -alkyl, heteroaryl, or heteroaryl-Cl-C 6 -alkyl 10 of which the cyclic moieties optionally may be substituted with one or more substitu ents independently selected from R .
183. A pharmaceutical composition according to claim 182 wherein R 55 is independently se lected from halogen, Cl-C 6 -alkyl, -CN, -OCF 3 ,-CF 3 , -NO 2 , -OR', -NR 56 R 57 , -NR 56 C(O)R 5 7 15 -SR 56 , -OC,-Cs-alkyl-C(O)OR" , or -C(O)OR" * C 1 -C 6 -alkyl optionally substituted with one or more substituents independently se lected from R 58 *ArG1, ArG 1-Cl-C 6 -alkyl, Het3, or Het3-C j -C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substitu 20 ents independently selected from R 59 .
184. A pharmaceutical composition according to claim 183 wherein R 55 is independently se lected from halogen, -OR 5 6 , -NRR 57 , -C(O)OR 56, -OC 1 -C 8 -alkyl-C(O)OR" , -NR"C(O)R 5 7 or CI-C 6 -alkyl.
185. A pharmaceutical composition according to claim 184 wherein R 55 is independently se 25 lected from halogen, -OR", -NR"R 7 , -C(O)OR 5 e, -OC-C 8 -alkyl-C(O)OR", -NR 56 C(O)R 57 , methyl or ethyl.
186. A pharmaceutical composition according to any one of the claims 173 to 185 wherein R5 and R 57 are independently selected from hydrogen, CF3, C 1 -C 1 2 -alkyl, or -C(=O)-Cl-C-alkyl; R5 and R 57 when attached to the same nitrogen atom may form a 3 to 8 30 membered heterocyclic ring with the said nitrogen atom.
187. A pharmaceutical composition according to claim 186 wherein R5 and R 5 7 are inde pendently selected from hydrogen or C 1 -C 12 -alkyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
188. A pharmaceutical composition according to claim 187 wherein R s 56 and R 57 are inde 35 pendently selected from hydrogen or methyl, ethyl, propyl butyl, R 56 and R s 57 when attached WO 2004/080480 PCT/DK2004/000158 454 to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro gen atom.
189. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is NN H N 5 AA wherein AA is Cl-C 6 -alkyl, aryl, heteroaryl, aryI-C1.6-alkyl- or aryl-C 2 -6-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 6 3 , and the aryl or heteroaryl is optionally substituted with one or more substituents 10 independently selected from R, R 63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 , R 64 is independently selected from 15 * hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -OS(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 65 , -NR 65 R 66 , -SR 65 , -NR 65 S(O) 2 R 66 , -S(O) 2 NR6R 66 , -S(O)NR 65 R 66 , -S(O)R 65 , -S(O) 2 R 65 , -OS(O) 2 R 65 , -C(O)NR 65 R 66 , -OC(O)NRs 65 R 66 , -NR 65 C(O)R 6 , -CH 2 C(O)NR 6 5 R 66 , -OC 1 ,-C 6 20 alkyl-C(O)NR 6 5 R 66 , -CH 2 0R 6 5 , -CH 2 0C(O)R 65 , -CH 2 NR 65 R 66 , -OC(O)R 65 , -OC1-C 6 alkyl-C(O)OR 65 , -OC,-C 6 -alkyl-OR 6 5 , -SC,-C6-alkyl-C(O)OR 65 , -C 2 -C 6 -alkenyl C(=O)OR 65 , -N R 65 -C(=O)-Cl-C 6 -alkyl-C(=O)OR 6 5 , -NR 65 -C(=O)-CI-C 6 alkenyl-C(=O)OR 65 , -C(O)OR 65 , or-C 2 -C 6 -alkenyl-C(=O)R 5 , 25 * C,-C 6 -alkyl, C 2 -C 6 -alkenyl or C 2 -Ce-alkynyl, each of which may optionally be substi tuted with one or more substituents selected from R 67 * aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C 1 -Ce-alkoxy, aryl-C 1 -C 6 -alkyl, aryl-C 2 -C 6 -alkenyl, aroyl-C 2 -C 6 -alkenyl, aryl-C 2 -C 6 -alkynyl, heteroaryl, heteroaryl-C, 30 C 6 -alkyl, heteroaryl-C 2 -C 6 -alkenyl or heteroaryl-C 2 -C 6 -alkynyl, WO 2004/080480 PCT/DK2004/000158 455 of which the cyclic moieties optionally may be substituted with one or more substitu ents selected from R 8 , R 6 5 and R5 6 are independently selected from hydrogen, OH, CF 3 , Cl-C 1 2 -alkyl, aryl-Cl-C6 5 alkyl, -C(=O)-R 69 , aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R 70 , and the aryl and heteroaryl groups may op tionally be substituted with one or more substituents independently selected from R 71 ; R 65 and R 66 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further 10 heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 6 7 is independently selected from halogen, -CN, -CF3, -OCF 3 , -OR 65 , and -NR 6 5 R 66 , 15 R 68 is independently selected from halogen, -C(O)OR 5 , -CH 2 C(O)OR 6 5 , -CH 2 0R 6 5 , -CN, CF 3 , -OCF 3 , -NO 2 , -OR 65 , -NR" 5 R 66 and Cl-C 6 -alkyl, R 69 is independently selected from Cl-C 6 -alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more C 1 -CG-alkyl, 20 R 70 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OC 1 -C 6 -alkyl, -C(O)OC 1 -C 6 alkyl, -COOH and -NH 2 , R 71 is independently selected from halogen, -C(O)OC 1 -C 6 -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , 25 NO 2 , -OH, -OC-C 6 -alkyl, -NH 2 , C(=O) or Cl-C 6 -alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
190. A pharmaceutical composition according to claim 189 wherein AA is aryl, heteroaryl or 30 aryl-C 1 .e-alkyl-, wherein the alkyl is optionally substituted with one or more R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 64.
191. A pharmaceutical composition according to claim 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently selected from R6. WO 2004/080480 PCT/DK2004/000158 456
192. A pharmaceutical composition according to claim 191 wherein AA is ArG1 or Hetl op tionally substituted with one or more substituents independently selected from R 4 .
193. A pharmaceutical composition according to claim 192 wherein AA is ArG1 or Het2 op tionally substituted with one or more substituents independently selected from R 64 5 194. A pharmaceutical composition according to claim 193 wherein AA is ArG1 or Het3 op tionally substituted with one or more substituents independently selected from R 64.
195. A pharmaceutical composition according to claim 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R 64. 10 196. A pharmaceutical composition according to claim 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently selected from R 64
197. A pharmaceutical composition according to any one of the claims 189 to 196 wherein R" is independently selected from hydrogen, halogen, -CF3, -OCF 3 , -OR 65 , -NR 6 6R 66 , C 1 -C 6 alkyl , -OC(O)R " 5 , -OC 1 -C 6 -alkyl-C(O)OR 5 , aryl-C 2 -C 6 -alkenyl, aryloxy or aryl, wherein C, 15 C 6 -alkyl is optionally substituted with one or more substituents independently selected from R 67 , and the cyclic moieties optionally are substituted with one or more substituents inde pendently selected from R 68 .
198. A pharmaceutical composition according to claim 197 wherein R6 is independently se lected from halogen, -CF 3 , -OCF 3 , -OR" 5 , -NR 65 R 66 , methyl, ethyl, propyl, -OC(O)R 6 5 , 20 -OCH 2 -C(O)OR 65 , -OCH 2 -CH 2 -C(O)OR 6 5 , phenoxy optionally substituted with one or more substituents independently selected from R 8 .
199. A pharmaceutical composition according to any one of the claims 189 to 198 wherein R 65 and R 66 are independently selected from hydrogen, CF 3 , C 1 -C 1 2 -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R" 1 . 25 200. A pharmaceutical composition according to claim 199 wherein R 65 and R 6 ' are inde pendently hydrogen, Cl-C 12 -alkyl, aryl, or heteroaryl optionally substituted with one or rnore substituents independently selected from R 71
201. A pharmaceutical composition according to claim 200 wherein R 65 and R 6 are inde pendently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Hetl optionally 30 substituted with one or more substituents independently selected from R 71 .
202. A pharmaceutical composition according to claim 201 wherein R 6 5 and R 66 are inde pendently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 7 . WO 2004/080480 PCT/DK2004/000158 457
203. A pharmaceutical composition according to claim 202 wherein R 6 5 and R 66 are inde pendently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 71 .
204. A pharmaceutical composition according to claim 203 wherein R 65 and R 66 are inde 5 pendently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadi azolyl optionally substituted with one or more R71 independently; or isoxazolyl optionally sub stituted with one or more substituents independently selected from R 71 .
205. A pharmaceutical composition according to any one of the claims 189 to 204 wherein R 71 is halogen or CI-C 6 -alkyl. 10 206. A pharmaceutical composition according to claim 205 wherein R 71 is halogen or methyl.
207. A pharmaceutical preparation according to any one of the claims 1 to 205 wherein Frgl consists of 0 to 5 neutral amino acids independently selected from the group consisting of Gly, Ala, Thr, and Ser.
208. A pharmaceutical preparation according to claim 207 wherein Frgi consists of 0 to 5 15 Gly.
209. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 0 Gly.
210. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 1 Gly.
211. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 2 Gly.
212. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 3 Gly. 20 213. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 4 Gly.
214. A pharmaceutical preparation according to claim 208 wherein Frgl consists of 5 Gly.
215. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein G" is of the formula B'-B 2 -C(O)-, B 1 -B 2 -SO 2 - or B 1 -B 2 -CH 2 -, wherein B' and B 2 are as defined in claim 1. 25 216. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein G B is of the formula B'-B 2 -C(O)-, B'-B 2 -SO2- or B'-B 2 -NH-, wherein B' and B 2 are as defined in claim 1.
217. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein G B is of the formula B'-B 2 -C(O)-, B 1 -B 2 -CH 2 - or B-B 2 -NH-, wherein B' and B 2 are as defined in 30 claim 1.
218. A pharmaceutical preparation according to any one of the claims 1 to 214 wherein GB is of the formula B'-B 2 -CH 2 -, 1-B 2 SO 2 - or B31-B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
219. A pharmaceutical preparation according to any one of the claims 215 or 216 wherein GB 35 is of the formula B 1 -B 2 -C(O)- or B'-B 2 -SO 2 -, wherein B' and B 2 are as defined in claim 1. WO 2004/080480 PCT/DK2004/000158 458
220. A pharmaceutical preparation according to any one of the claims 215 or 217 wherein GB is of the formula B'-B 2 -C(O)- or B'-B 2 -CH 2 -, wherein B 1 and B 2 are as defined in claim 1.
221. A pharmaceutical preparation according to any one of the claims 216 or 217 wherein GB is of the formula B 1 -B 2 -C(O)- or B'-B 2 -NH-, wherein B' and B 2 are as defined in claim 1. 5 222. A pharmaceutical preparation according to any one of the claims 215 or 218 wherein GB is of the formula B'-B 2 -CH 2 - or B'-B 2 -SO 2 -, wherein B' and B 2 are as defined in claim 1.
223. A pharmaceutical preparation according to any one of the claims 216 or 218 wherein GB is of the formula B'-B 2 -NH- or B'-B 2 -SO 2 -, wherein B' and B 2 are as defined in claim 1.
224. A pharmaceutical preparation according to any one of the claims 217 or 218 wherein GB 10 is of the formula B'-B 2 -CH 2 - or B 1 -B 2 -NH-, wherein B' and B 2 are as defined in claim 1.
225. A pharmaceutical preparation according to any one of the claims 219, 220, or 221 wherein GB is of the formula B'-B 2 -C(O)-.
226. A pharmaceutical preparation according to any one of the claims 220, 222 or 224 wherein GB is of the formula B'-B 2 -CH 2 -. 15 227. A pharmaceutical preparation according to any one of the claims 220, 222 or 223 wherein G3 is of the formula B'-B 2 -SO2-.
228. A pharmaceutical preparation according to any one of the claims 221, 223 or 224 wherein G3 is of the formula B'-B 2 -NH-.
229. A pharmaceutical preparation according to any one of the claims 1 to 228 wherein B' is 20 a valence bond, -0-, or -S-.
230. A pharmaceutical preparation according to any one of the claims 1 to 228 wherein B' is a valence bond, -0-, or -N(RB)-.
231. A pharmaceutical preparation according to any one of the claims 1 to 228 wherein B' is a valence bond, -S-, or -N(RB) -. 25 232. A pharmaceutical preparation according to any one of the claims 1 to 228 wherein B 1 is -0-, -S- or -N(R 6 B)-.
233. A pharmaceutical preparation according to any one of the claims 229 or 230 wherein B 1 is a valence bond or -0-.
234. A pharmaceutical preparation according to any one of the claims 229 or 231 wherein B' 30 is a valence bond or -S-.
235. A pharmaceutical preparation according to any one of the claims 230 or 231 wherein B' is a valence bond or -N(R6B)_.
236. A pharmaceutical preparation according to any one of the claims 229 or 232 wherein B' is -O-or -S-. WO 2004/080480 PCT/DK2004/000158 459
237. A pharmaceutical preparation according to any one of the claims 230 or 232 wherein B' is -O-or -N(R 6 ")-.
238. A pharmaceutical preparation according to any one of the claims 231 or 232 wherein B' is -S-or -N(R 6 B)-. 5 239. A pharmaceutical preparation according to any one of the claims 233, 234 or 235 wherein B' is a valence bond.
240. A pharmaceutical preparation according to any one of the claims 233, 236 or 237 wherein B' is -0-.
241. A pharmaceutical preparation according to any one of the claims 234, 236 or 238 10 wherein B' is -S-.
242. A pharmaceutical preparation according to any one of the claims 235, 237 or 238 wherein B' is -N(R6B) - .
243. A pharmaceutical preparation according to any one of the claims 1 to 242 wherein B 2 is a valence bond, C,-C, 8 -alkylene, C 2 -C1 8 -alkenylene, C 2 -C,1 8 -alkynylene, arylene, heteroary 15 lene, -C,-C, 8 -alkyl-aryl-, -C(=0)-C-Cj8-alkyl-C(=O)-, -C(=0)-Cl-C 8 -alkyl-O-Cl-C, 8 -alkyl C(=O)-, -C(=O)-C1-Cl8-alkyl-S-Cl-C,8-alkyl-C(=O)-, -C(=O)-Cl-Cl8-alkyl-NR6-Cl-C18-alkyl C(=O)-; and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
244. A pharmaceutical preparation according to claim 243 wherein B 2 is a valence bond, C, 20 C1 8 -alkylene, C 2 -C 18 -alkenylene, C 2 -C18-alkynylene, arylene, heteroarylene, -Cl-C 18 -alkyl-aryl , -C(=O)-C,-Cj 8 -alkyl-C(=O)-, -C(=O)-Cl-Cs, 8 -alkyl-O-Cl-Cl 8 -alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
245. A pharmaceutical preparation according to claim 244 wherein B 2 is a valence bond, C, C1 8 -alkylene, C 2 -C1 8 -alkenylene, C 2 -C 1 8 -alkynylene, arylene, heteroarylene, -C,-C, 8 -alkyl-aryl 25 , -C(=O)-C1-C 8 -alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substi tuted as defined in claim 1.
246. A pharmaceutical preparation according to claim 245 wherein B 2 is a valence bond, Cj C1 8 -alkylene, arylene, heteroarylene, -Cl-C 8 -alkyl-aryl-, -C(=0)-Cj-C1 8 -alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. 30 247. A pharmaceutical preparation according to claim 246 wherein B 2 is a valence bond, C 1 C1 8 -alkylene, arylene, heteroarylene, -Cl-C, 8 -alkyl-aryl-, and the alkylene and arylene moie ties are optionally substituted as defined in claim 1.
248. A pharmaceutical preparation according to claim 247 wherein B 2 is a valence bond, C, C1 8 -alkylene, arylene, -Cl-C, 8 -alkyl-aryl-, and the alkylene and arylene moieties are optionally 35 substituted as defined in claim 1. WO 2004/080480 PCT/DK2004/000158 460
249. A pharmaceutical preparation according to claim 248 wherein B 2 is a valence bond or -Cl-C 1 8 -alkylene, and the alkylene moieties are optionally substituted as defined in claim 1.
250. A pharmaceutical preparation according to any one of the claims 1 to 249 whereiin Frg2 comprises 1 to 16 positively charged groups. 5 251. A pharmaceutical preparation according to claim 250 wherein Frg2 comprises 1 to 12 positively charged groups.
252. A pharmaceutical preparation according to claim 251 wherein Frg2 comprises 1 to 10 positively charged groups.
253. A pharmaceutical preparation according to any one of the claims 1 to 249 wherein Frg2 10 comprises 10 to 20 positively charged groups.
254. A pharmaceutical preparation according to claim 253 wherein Frg2 comprises 12 to 20 positively charged groups.
255. A pharmaceutical preparation according to claim 254 wherein Frg2 comprises 16 to 20 positively charged groups. 15 256. A pharmaceutical preparation according to any one of the claims 250 to 255 wherein the positively charged groups of Frg2 are basic amino acids independently selected from the group consisting of Lys and Arg and D-isomers of these.
257. A pharmaceutical preparation according to claim 256 wherein the basic amino acids are all Arg. 20 258. A pharmaceutical preparation according to any one of the claims 250 to 257, wherein Frg2 comprises one or more neutral amino acids independently selected from the group con sisting of Gly, Ala, Thr, and Ser.
259. A pharmaceutical preparation according to claim 258, wherein Frg2 comprises one or more Gly. 25 260. A pharmaceutical preparation according to any one of the claims 1 to 259 wherein X is OH or -NH 2 .
261. A pharmaceutical preparation according to claim 260 wherein X is -NH 2 .
262. A pharmaceutical preparation according to any one of the claims 1 to 261 which further comprises at least 3 phenolic molecules per putative insulin hexamer. 30 263. A pharmaceutical preparation according to any one of the claims 1 to 262 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gln, Glu, Gly, His, lie, Leu, Met, Phe, Ser, Thr, Trp, Tyr, Val, and hSer.
264. A pharmaceutical preparation according to claim 263 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala, Gly, lie, Leu, Phe, Ser, Thr, Val, and 35 hSer. WO 2004/080480 PCT/DK2004/000158 461
265. A pharmaceutical preparation according to claim 264 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Ala or Gly.
266. A pharmaceutical preparation according to claim 265 wherein the acid-stabilised insulin is an analogue of human insulin wherein A21 is Gly. 5 267. A pharmaceutical preparation according to any one of the claims 263 to 266 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange or dele tion of one or more amino acid residues according to the following: B3 is selected from Thr, Ser, Lys or Ala A18 is Gin 10 B28 is Lys, Asp or Glu B29 is Pro or Glu B9 is Glu or Asp 810 is Glu B25 is deleted 15 B30 is deleted.
268. A pharmaceutical preparation according to claim 267 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys or Asp.
269. A pharmaceutical preparation according to claim 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Asp. 20 270. A pharmaceutical preparation according to claim 268 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B28 to Lys.
271. A pharmaceutical preparation according any one of the claims 263 to 270 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B29 to Pro. 25 272. A pharmaceutical preparation according any one of the claims 263 to 271 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of B3 to Lys or Ala.
273. A pharmaceutical preparation according any one of the claims 263 to 272 wherein the acid-stabilised insulin is an analogue of human insulin further modified by exchange of A 18 30 to Gln.
274. A pharmaceutical preparation according any one of the claims 263 to 273 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B25.
275. A pharmaceutical preparation according any one of the claims 263 to 274 wherein the acid-stabilised insulin is an analogue of human insulin further modified by deletion of B30. WO 2004/080480 PCT/DK2004/000158 462
276. A pharmaceutical preparation according to claim 263 wherein the acid-stabilised insulin is selected from the group A21G A21G, B28K, B29P 5 A21G,B28D A21G, B28E A21G, B3K, B29E A21G, desB27 A21G, B9E 10 A21G, B9D A21G, B10E A21G, desB25 A21G, desB30 A21G, B28K, B29P, desB30 15 A21G, B28D, desB30 A21G, B28E, desB30 A21G, B3K, B29E, desB30 A21G, desB27, desB30 A21G, B9E, desB30 20 A21G, B9D, desB30 A21G, B10E, desB30 A21G, desB25, desB30.
277. A pharmaceutical preparation according to any one of the claims 1 to 276 wherein zinc ions are present in an amount corresponding to 10 to 40 pg Zn/100 U insulin. 25 278. A pharmaceutical preparation according to claim 277 wherein zinc ions are present in an amount corresponding to 10 to 26 pg Zn/100 U insulin.
279. A pharmaceutical preparation according to any one of the claims 1 to 278 wherein the ratio between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 99:1 to 1:99. 30 280. A pharmaceutical preparation according to claim 279 wherein the ratio between insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 95:5 to 5:95.
281. A pharmaceutical preparation according to claim 280 wherein the ratio between be tween insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the 35 range from 80:20 to 20:80. WO 2004/080480 PCT/DK2004/000158 463
282. A pharmaceutical preparation according to claim 281 wherein the ratio between be tween insulin and the zinc-binding ligand according to any one of the claims 1 to 261 is in the range from 70:30 to 30:70.
283. A pharmaceutical preparation according to any one of the claims 1 to 282 wherein the 5 concentration of insulin is 60 to 3000 nmol/ml.
284. A pharmaceutical preparation according to claim 283 wherein the concentration of insulin is 240 to 1200 nmol/ml.
285. A pharmaceutical preparation according to claim 284 wherein the concentration of insulin is about 600 nmol/ml. 10 286. A method of preparing a zinc-binding ligand according to claim 1 comprising the steps of * Identifying starter compounds that binds to the R-state HisBlO-Zn 2 + site *optionally attaching a fragment consisting of 0 to 5 neutral a- or 1-amino acids * attaching a fragment comprising 1 to 20 positively charged groups independently se 15 lected from amino or guanidino groups.
287. Method of prolonging the action of an acid-stabilised insulin preparation which com prises adding a zinc-binding ligand according to any of claims 1 to 261 to the acid-stabilised insulin preparation.
288. A method of treating type 1 or type 2 diabetes comprising administering to a patient in 20 need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the claims 1 to 282.
289. Use of a preparation according to any one of the claims 1 to 282 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
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