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WO2002004458A1 - Modulators of protein tyrosine phosphatases (ptpases) - Google Patents

Modulators of protein tyrosine phosphatases (ptpases) Download PDF

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Publication number
WO2002004458A1
WO2002004458A1 PCT/DK2001/000450 DK0100450W WO0204458A1 WO 2002004458 A1 WO2002004458 A1 WO 2002004458A1 DK 0100450 W DK0100450 W DK 0100450W WO 0204458 A1 WO0204458 A1 WO 0204458A1
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WO
WIPO (PCT)
Prior art keywords
amino
aryl
methyl
carboxylic acid
thieno
Prior art date
Application number
PCT/DK2001/000450
Other languages
French (fr)
Inventor
Thomas Kruse Hansen
Jesper Lau
Niels Peter Hundahl Møller
Ole Hvilsted Olsen
Frank Urban Axe
Farid Bakir
Yu Ge
Daniel Dale Holsworth
Luke Milburn Judge
Michael James Newman
Roy Teruyuki Uyeda
Barry Zvi Shapira
Henrik Sune Andersen
Original Assignee
Novo Nordisk A/S
Ontogen Corporation
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Filing date
Publication date
Application filed by Novo Nordisk A/S, Ontogen Corporation filed Critical Novo Nordisk A/S
Priority to AU2001268951A priority Critical patent/AU2001268951A1/en
Publication of WO2002004458A1 publication Critical patent/WO2002004458A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

Definitions

  • the present invention relates to novel thienopyridines, to methods for their preparation, to compositions comprising the compounds, to the use of these compounds as medicaments and their use in therapy, where such compounds of Formula 1 are pharmacologically useful inhibitors or modulators of Protein Tyrosine Phosphatases (PTPases) including PTP1 B and T cell PTP (TC-PTP),
  • PTPases Protein Tyrosine Phosphatases
  • TC-PTP T cell PTP
  • PTPases play a major role in the modulation and regulation of fundamental cellular signaling mechanisms involved in metabolism, growth, proliferation and differentiation (Fischer et al, Science 253:401-6 (1991); Tonks and Neel, Cell 87: 365-368 (1996); Neel and Tonks, Current Opinion in Cell Biology 9: 193-204 (1997); Hunter, Phil. Trans. R. Soc. Lond. B 353: 583- 605 (1998); Zhang, Critical Reviews in Biochemistry and Molecular Biology 33:1- 52 (1998)).
  • PTPase inhibitors may help treat or manage certain types of diseases such as type 1 and type 2 diabetes, obesity, autoimmune diseases, acute and chronic inflammation, osteoporosis and various forms of cancer.
  • certain infectious diseases may also be treated or managed by administration PTPase inhibitors (Clemens et al. , Molecular Microbiology 5: 2617-2620 (1991 )).
  • Protein phosphorylation is now well recognized as an important mechanism utilized by cells to transduce and regulate signals during different stages of cellular function (Hunter, vide supra; Chan et al., Annu. Rev. Immunol. 12: 555- 592 (1994); Zhang, Curr. Top. Cell. Reg. 35: 21-68 (1997); Matozaki and Kasuga, Cell. Signal. 8: 113-19 (1996); Fischer et al, vide supra).
  • the level of tyrosine phosphorylation is balanced by the opposing action of protein tyrosine kinases and protein tyrosine phosphatases (PTPases).
  • phosphatases There are at least two major classes of phosphatases: (1) those that dephosphorylate proteins (or peptides) that contain a phosphate group(s) on a serine or threonine moiety (termed Ser/Thr phosphatases) and (2) those that remove a phosphate group(s) from the amino acid tyrosine (termed protein tyrosine phosphatases or PTPases or PTPs).
  • the PTPases are a family of enzymes that can be classified into two groups: a) intracellular or nontransmembrane PTPases and b) receptor-type or transmembrane PTPases.
  • dual-specificity phosphatases and low molecular weight phosphatases can also dephosphorylate phosphotyrosyl proteins (WO97/39748, WO97/40017, WO99/15529, WO97/08934, WO98/27065, WO99/46236, WO99/46244, WO99/46267, WO99/46268, WO99/46237).
  • PTPases play a major role in the above modulation and regulation of fundamental cellular signaling mechanisms involved in metabolism, growth, proliferation and differentiation (Fischer et al, Science 253:401-6 (1991); Tonks and Neel, Ce// 87: 365-368 (1996); Neel and Tonks, Current Opinion in Cell Biology 9: 193-204 (1997); Hunter, Phil. Trans. R. Soc. Lond. B 353: 583-605 (1998); Zhang, Critical Reviews in Biochemistry and
  • PTPases can act both as positive and negative regulators of signal transduction processes.
  • PTPases have been implicated in a variety of human diseases, including type 1 and type 2 diabetes, obesity, autoimmune diseases, acute and chronic inflammation, osteoporosis, proliferative disorders including various forms of cancer, growth disorders, and defective platelet aggregation (WO97/39748, WO97/40017, WO99/15529, WO97/08934, WO98/27065, WO99/46236, WO99/46244, WO99/46267, WO99/46268, WO99/46237).
  • Both selective PTPase inhibitors and inhibitors that bind to several PTPases can be used therapeutically to partially or completely restore PTPase-mediated perturbed signal transduction processes and thus for management, treatment or prevention of the above diseases.
  • WO 99/46267 discloses compounds, which are pharmacologically useful inhibitors of PTPases.
  • the present invention which represents a novel selection under WO 99/46267, discloses a class of compounds, which surprisingly are more potent against protein tyrosine phosphatases (e.g. PTP1B) than those disclosed in WO 99/46267.
  • the present invention relates to compounds of the Formula 1 wherein X, R ⁇ R 2 , R 3 , and R are defined below;
  • X is -C(O)- or -S(O) 2 -;
  • R ! and R 2 are independently hydrogen, C C 6 alkyl, aryl-R 5 -, R 6 -C(O)-O-R 7 - or aryl-R 8 -C(O)-O-R 9 - wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, C C ⁇ alkyl or C C 6 alkyloxy;
  • R 3 is hydrogen, C Cealkyl, H 2 N-R 37 -, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, aryl, aryl-R ⁇ 0 -, aryl-N(R35)-, aryl-R 11 -N(R 36 )- , C C 6 alkyloxy or aryl-R 13 -O- where
  • R 4 is hydrogen, C C 6 alkyl, aryl-R 26 -, R 27 -O-C(O)-, aryl-R 28 -O-C(O)-, R 29 -C(O)-O- R 30 -O-C(O)- or aryl-R 3 C(O)-O-R 32 -O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, aryl, aryl-R 33 -, C C 6 aIkyloxy or aryl-R 34 -O- and wherein aryl group is phenyl, naphthyl or thiophenyl; and wherein R5, R 7 , R 8 , Rg, R10, Rn, R ⁇ 3> R ⁇ 4 , R15.
  • R 2 3 > R 2 ⁇ .
  • R3i > R3 2 .
  • R 25 , R 27 and R 29 independently are C 1-6 alkyl and wherein R 6 , R ⁇ 9 , R 2 ⁇ , R 24 , R 35 ,
  • R 36 and R 38 independently are hydrogen or C 1-6 alkyl; 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 f tautomeric form.
  • the compounds of the invention can be further modified to act as prodrugs.
  • a preferred prodrug is acetoxymethyl esters or acetoxymethyl carbamates of the compounds of the present invention.
  • acetoxymethyl esters or acetoxymethyl carbamates of the compounds of the present invention As a general procedure preparation of an acetoxymethyl ester is given below (C.Schultz et al, The Journal of Biological Chemistry, 1993, 268, 6316-6322.;.-
  • attachment or "-" signifies a stable covalent bond, certain preferred points of attachment points being apparent to those skilled in the art.
  • halogen and halo includes fluorine, chlorine, bromine, and iodine.
  • alkyl includes C ⁇ Ce straight chain saturated, C C 6 branched chain saturated and C 3 -C 6 cyclic saturated hydrocarbon groups.
  • this definition shall include but is not limited to methyl (Me), ethyl (Et), propyl (Pr), butyl (Bu), pentyl, hexyl, isopropyl (i-Pr), isobutyl (i-Bu), fe/f-butyl (t- u), sec-butyl
  • alkenyl includes C 2 -C 6 unsaturated aliphatic hydrocarbon groups and
  • C 2 -C 6 branched unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms and at lest one double bond.
  • this definition shall include but is not limited to ethenyl, propenyl, butenyl, pentenyl, hexenyl, isopentenyl, neopentenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like.
  • alkynyl includes C 2 -C 6 straight chain unsaturated aliphatic, C 2 -C 6 branched unsaturated and cyclic C 6 unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms and at lest one triple bond.
  • this definition shall include but is not limited to acetynyl, propynyl, butynyl, pentynyl, hexynyl, cyclohexynyl and the like.
  • alkyloxy (e.g. methoxy, ethoxy, propyloxy, allyloxy, cyclohexyloxy) represents an "alkyl” group as defined above having the indicated number of carbon atoms attached through an oxygen bridge.
  • aryloxy e.g. phenoxy, naphthyloxy and the like
  • aryloxy represents an aryl group as defined below attached through an oxygen bridge.
  • aryl represents an unsubstituted, mono-, di- or trisubstituted monocyclic, polycyclic, biaryl or heterocyclic aromatic group(s) covalently attached at any ring position capable of forming a stable covalent bond, certain preferred points of attachment being apparent to those skilled in the art (e.g., 3- indolyl, 4(5)-imidazolyl).
  • aryl includes phenyl, biphenyl, indenyl, naphthyl (1 -naphthyl, 2- naphthyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1 ,2,3-triazoM-yl, 1 ,2,3-triazol-2-yl 1 ,2,3-triazol-4-yl, 1 ,2,4-triazol-3-yl), thiophenyl (2-thiophenyl, 3-thiophenyl, 4-thiophenyl, 5-thiophenyl), pyridyl (2- pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl,
  • modified compounds which are not intended in any way to limit the scope of the invention, are compounds that have been, cyclized at specific positions - so called 'cyclic compounds' - which upon uptake in cells or mammals become hydrolyzed at the same specific position(s) in the molecule to yield the compounds of the invention, the original compounds, which are then said to be 'non-cyclic'.
  • the latter original compounds in most cases will contain other cyclic or heterocyclic structures that will not be hydrolyzed after uptake in cells or mammals.
  • said modified compounds may not show behavior in biochemical assays similar to that of the original compound, i.e. the corresponding compounds of the invention without the attached chemical groups or said modifications.
  • Said modified compounds may even be inactive in biochemical assays. However, after uptake in cells or mammals these attached chemical groups of the modified compounds may in turn be removed spontaneously or by endogenous enzymes or enzyme systems to yield compounds of the invention, original compounds. 'Uptake' is defined as any process that will lead to a substantial concentration of the compound inside cells or in mammals. After uptake in cells or mammals and after removal of said attached chemical group or hydrolysis of said cyclic compound, the compounds may have the same structure as the original compounds and thereby regain their activity and hence become active in cells and/or in vivo after uptake.
  • the term 'a functional group which can be converted to hydrogen in vivo' is intended to include any group which upon administering the present compounds to the subjects in need thereof can be converted to hydrogen e.g. enzymatically or by the acidic environment in the stomach.
  • the compounds of the present invention have asymmetric centres and may occur as racemates, racemic mixtures, and as individual enantiomers or diastereoisomers, with all isomeric forms being included in the present invention as well as mixtures thereof.
  • salts of the compounds of Formula 1 where a basic or acidic group is present in the structure, are also included within the scope of this invention.
  • an acidic substituent such as -COOH, 5- tetrazolyl or -P(O)(OH) 2 ⁇ there can be formed the ammonium, morpholinium, sodium, potassium, barium, calcium salt, and the like, for use as the dosage form.
  • an acidic salt such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate, ethanesulfonate, ethane sulfonate, picrate and the like, and include acids related to the pharmaceutically acceptable salts listed in Journal of Pharmaceutical Science, 66, 2 (1977) and incorporated herein by reference, can be used as the dosage form.
  • an acidic salt such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate,
  • esters can be employed, e.g., methyl, tert-butyl, pivaloyloxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodrug formulations.
  • some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.
  • terapéuticaally effective amount shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or other.
  • the present invention is concerned with compounds of Formula 1
  • Ri and R 2 are independently hydrogen, d-C 6 alkyl, aryl-R 5 -, R 6 -C(O)-O-R 7 - or aryl-R 8 -C(O)-O-R 9 - wherein aryl is phenyl, naphthyl, thiophenyl, which aryl group is optionally substituted with halogen, trihalomethyl, aryl, aryl- , Rio-, C C 6 alkyloxy or aryl-R ⁇ O-;
  • R 3 is CrC 6 alkyl, H 2 N-R 35 -, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, aryl-R ⁇ 2 -, aryl, aryl-R 13 -, aryl-N(R ⁇ 4 )-, aryl-R 5 -N(R 16 )-, R 38 -aryl-, C C 6 alkyloxy or aryl-R ⁇
  • R 2 o, R 22 , R 2 3, R 24 , R 2 5, R 2 6, and R 35 independently are d-C ⁇ -alkylene, wherein R ⁇ 2 is alkenylene, wherein R 6 , R 2 ⁇ , and R 38 independently are CrC 6 alkyl and wherein R 14 and R 16 independently are hydrogen or C ⁇ -6 alkyl;
  • More preferred compounds of the invention are compounds of Formula 1
  • X is -C(O)-
  • Ri and R 2 are independently hydrogen or C C 6 alkyl
  • R 3 is CrC 6 alkyl, H 2 N-R 18 , aryl-R 5 -, aryl, aryl-R 6 -, aryl-N(R 7 )-, aryl-R 8 -O- or R 19 - aryl- wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, or indolyl, which aryl group is optionally substituted with halogen, hydroxy, CrCealkyl, aryl, aryl-R 9 -, C C 6 alkyloxy, aryloxy, aryl-R 10 -O-;
  • R 4 is hydrogen, aryl-Rn-, R ⁇ 2 -C(O)-O-R 13 -O-C(O)- or aryl-R ⁇ -C(O)-O-R 15
  • R3 is CrC 6 alkyl, H 2 N-R ⁇ 2 -, aryl, aryl-R 5 -, aryl-N(R 6 )-, aryl-R 7 -O- or R 13 -aryl-, wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, or indolyl, which aryl group is optionally substituted with halogen, hydroxy, d-Cealkyl, aryl, aryl-R 8 -, CrC 6 alkyloxy, aryloxy, aryl-R 9 -O- wherein aryl is phenyl or thiophenyl, which aryl group is optionally substituted with halogen, hydroxy, nitro, cyano, trihalomethyl, aryl, aryl-R 10 -, Cr
  • any optical isomer or mixture of optical isomers including a racemic mixture, or any tautomeric form.
  • the invention relates to a compound of Formula 1
  • X is -C(O)- or -S(O) 2 -;
  • Ri and R 2 are independently hydrogen or a functional group that can be converted to hydrogen in vivo;
  • R 3 is hydrogen, CrC 6 alkyl, H 2 N-R 35 -, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, aryl, aryl-R ⁇ 0 -, aryl-N(R 35 )-, aryl-Rn-N(R 36 )- , d-C 6 alkyloxy or aryl-R 13 -O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC 6 alkyl, R 37 -aryl-, R 38 -, R 38 - R 14 -, CrCealky
  • R 4 is hydrogen, CrC 6 alkyl, aryl-R 26 -, R 27 -O-C(O)-, aryl-R 28 -O-C(O)-, R 29 -C(O)-O- R 30 -O-C(O)- or aryl-R 3 rC(O)-O-R 32 -O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R 39 -, R 39 -R 33 -, C C 6 alkyloxy or R 39 -R 34 -O-; and wherein R5, R 7 , R 8 , R 9 , R 0, Rn, R13, R ⁇ 4 , R15, R 2 3.
  • R and R 2 are independently hydrogen, C ⁇ -C 6 alkyl, aryl- R 5 -, R 6 -C(O)-O-R 7 - or aryl-R 8 -C(O)-O-R 9 - wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, CrC 6 alkyl or d-C 6 alkyloxy;
  • X is C(O).
  • X is S(O) 2 .
  • R 1 and R 2 are independently hydrogen, CrC 6 alkyl, aryl- R -, or R 6 -C(O)-O-R 7 -, wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, d-C 6 alkyl or Ci- Cealkyloxy.
  • Ri and R 2 are independently hydrogen or d-Cealkyl.
  • Ri and R 2 are hydrogen.
  • R 3 is C C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, aryl, aryl- R 1 0-, CrC 6 alkyloxy, H 2 N-R 35 - or aryl-R ⁇ 3 -O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC 6 alkyl, R 37 -aryl-, R 38 -, R 38 - Ri 4 -, CrCealkyloxy, Ras-O-, R 38 -R ⁇ 5 -O-, R3 8 -N(R ⁇ 6
  • R 3 is CrC 6 alkyl, C 2 -C 6 alkenyl, aryl, H 2 N-R 35 - or aryl-R 13 - O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl , group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrCealkyl, R 37 -aryl-, R 38 -, R 38 -Ri4-, d-C 6 alkyloxy, R 38 -O-, R 38 -R ⁇ 5 -O-, Rs 8 -N(R ⁇ 6 )- , R ⁇ 8 -C(O)-N(R ⁇ 9 )-, R 38
  • R 3 is d-C 6 alkyl or aryl, wherein aryl is phenyl, biphenyl, . indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC 6 alkyl, R 37 -aryl-, R 38 -, R 38 - R ⁇ 4 -, CrCealkyloxy, R 38 -O-, R 38 -R ⁇ 5 -O-, R 38 -N(R ⁇ 6 )-, R ⁇ 8 -C(O)-N(R ⁇ 9 )-, R 38 -C(O)- N(R 2 ⁇ )- or R 38 -R 23 -C(O)-N
  • R 3 is aryl, wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC 6 alkyl, R 37 -aryl-, R 38 -, R ⁇ -R ⁇ -, Ci- Cealkyloxy, R 38 -O-, R 38 -Ri 5 -O-, R 38 -N(R ⁇ 6 )-, R ⁇ 8 -C(O)-N(R ⁇ 9 )-, R 38 -C(O)-N(R 2 ⁇ )- or R 38 -R 23 -C(O)-N(R 24 )-.
  • aryl is pheny
  • aryl of R 3 is phenyl, naphthyl, or indolyl.
  • aryl of R 3 is phenyl
  • the aryl of R 3 is substituted by halogen, hydroxy, d- C 6 alkyl, R 37 -aryl-, R 38 -, R 38 -R ⁇ -, CrC 6 alkyloxy, R 38 -O-, R 38 -R ⁇ 5 -O-, R ⁇ 8 -C(O)- N(R ⁇ 9 )-, or R 38 -C(O)-N(R 2 ⁇ )-.
  • the aryl of R 3 is substituted by hydroxy, CrC 6 alkyl, R 38 -, R 38 -R ⁇ -, CrCealkyloxy, R 38 -O-, R ⁇ 8 -C(O)-N(R ⁇ 9 )-, or R 38 -C(O)-N(R 2 ⁇ )-.
  • R 38 is phenyl or thiophenyl.
  • R 4 is hydrogen, CrC 6 alkyl, aryl-R 26 -, R 29 -C(O)-O-R 30 -O-, C(O)- or aryl-R 3 rC(O)-O-R 32 -O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R 39 -, R 39 -R 33 -, CrCealkyloxy or R 39 -R 34 -O-.
  • R 4 is hydrogen, CrC 6 alkyl, or aryl-R 26 - wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R 39 -, R 39 -R 33 -, CrC 6 alkyloxy or R 39 -R 34 -O-.
  • R is hydrogen or aryl-R 26 - wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R 39 -, R 39 -R 33 -, CrC 6 alkyloxy or R 39 -R 34 -O-.
  • R 4 is hydrogen
  • R 4 is aryl-R 26 - wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R 39 -, R3 9 -R 33 -, d-C 6 alkyloxy or R 39 -R 3 -O-.
  • aryl of R 4 is phenyl. In another embodiment the aryl of R 4 is substituted by R ⁇ -.
  • R 39 is CrC 6 alkyloxy.
  • R 39 is methoxy
  • Another aspect of the invention is compounds according to the invention that act as inhibitors of Protein Tyrosine Phosphatases.
  • Another aspect of the invention is a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of the invention or a pharmaceutically acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents.
  • Another aspect of the invention is a pharmaceutical composition suitable for treating type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity comprising a compound of the invention or a pharmaceutically acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents.
  • Another aspect of the invention is a pharmaceutical composition suitable for treating immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases comprising a compound of the invention or a pharmaceutical acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents.
  • the composition may be in the form of an oral dosage unit or parenteral dosage unit.
  • the pharmaceutical composition may said may contain a compound of the invention such that it is administered as a dose in a range from about 0.05 to 1000 mg, preferably from about 0.1 to 500 mg and especially in the range from 50 to 200 mg per day
  • Another aspect of the invention is a compound of the invention or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable aqid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric form for therapeutical use in the treatment or , prevention of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity.
  • Another aspect of the invention is a compound of the invention or a pharmaceutically acceptable 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 form for therapeutical use in the treatment or preventing of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases.
  • immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or
  • Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form as a medicament.
  • Another aspect of the invention is the use of a compound of the invention for preparing a medicament.
  • Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form for the preparation of a medicament suitable for the treatment or preventing of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity.
  • Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form for the preparation of a medicament suitable for the treatment or preventing of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases.
  • immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines
  • Another aspect of the invention is a method of treating type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity comprising administering to a subject in need thereof an effective amount of a compound of the invention.
  • Another aspect of the invention is a method of treating immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases comprising administering to a subject in need thereof an effective amount of a compound of the invention to said subject.
  • Another aspect of the invention is a process for the manufacture of a medicament, particular to be used in the treatment or prevention of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity which process comprising bringing a compound of the invention or a pharmaceutically acceptable salt thereof into a galenic dosage form.
  • Another aspect of the invention is a process for the manufacture of a medicament, particular to be used in the treatment or prevention of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases which process comprising bringing a compound of the invention or a pharmaceutically acceptable salt thereof into a galenic dosage form.
  • Another aspect of the invention is a method for preparing a compound of formula
  • the compounds are evaluated for biological activity with a truncated form of PTP1 B (corresponding to the first 321 amino acids), which was expressed in E. coli and purified to apparent homogeneity using published procedures well- known to those skilled in the art.
  • the enzyme reactions are carried out using standard conditions essentially as described by Burke et al. (Biochemistry 35; 15989-15996 (1996)).
  • the assay conditions are as follows. Appropriate concentrations of the compounds of the invention are added to the reaction mixtures containing different concentrations of the substrate, p-nitrophenyl phosphate (range: 0.16 to 10 mM - final assay concentration).
  • the buffer used was 100 mM sodium acetate pH 5.5, 50 mM sodium chloride, 0.1 % (w/v) bovine serum albumin and 5 mM dithiothreitol (total volume 100 ml).
  • the reaction was started by addition of the enzyme and carried out in microtiter plates at 25 °C for 60 minutes. The reactions are stopped by addition of NaOH.
  • the enzyme activity was determined by measurement of the absorbance at 405 nm with appropriate corrections for absorbance at 405 nm of the compounds and p-nitrophenyl phosphate.
  • the data are analyzed using nonlinear regression fit to classical Michaelis Menten enzyme kinetic models. Inhibition is expressed as K
  • Table 1 The results of representative experiments are shown in Table 1.
  • reaction step a) in Method A gives a mixture of regioisomers which can be separated by use of column chromatography known to thus skilled in the art.
  • C(CO)- and W is -OH, -OSO 2 Me, halogen, R 4 COO- or X is -SO 2 - and W is chloride, a substituted 5-aminomethyl-tetrahydro-thieno[2,3-c]pyridine (II) to react under conditions known to thus skilled in the art which favour amide or sulfon amide bond formation followed by b) R 2 -O-C(O)-C(O)-imidazol-1-yl, tetrahydrofuran and c) 25% trifluoroacetic acid/dichloromethane; to yield (III) wherein R 1 ( R 2 , R 3 , and R 4 are defined above;
  • dosages suitable for oral administration comprise from about 0.5 mg to about 1000 mg, preferably , from about 1 mg to about 500 mg of the compounds of the invention admixed with a pharmaceutical carrier or diluent.
  • the compounds of the invention may be administered in a pharmaceutically acceptable acid addition salt form or where possible as a metal or a - ⁇ - alkylammonium salt.
  • Such salt forms exhibit approximately the same order of activity as the free acid forms.
  • compositions comprising a compound of the invention or a pharmaceutically acceptable salt thereof and, usually, such compositions also contain a pharmaceutical carrier or diluent.
  • compositions containing the compounds of this invention may be prepared by conventional techniques and appear in conventional forms, for example capsules, tablets, solutions or suspensions.
  • the pharmaceutical carrier employed may be a conventional solid or liquid carrier.
  • solid carriers are lactose, terra alba, sucrose, talc, gelatine, agar, pectin, acacia, magnesium stearate and stearic acid.
  • liquid carriers are syrup, peanut oil, olive oil and water.
  • the carrier or diluent may include any time delay material known to the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. If a solid carrier for oral administration is used, the preparation can be tabletted, placed in a hard gelatine capsule in powder or pellet form or it can be in the form of a troche or lozenge.
  • the amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
  • the compounds of this invention are dispensed in unit dosage form comprising 10-200 mg of active ingredient in or together with a pharmaceutically acceptable carrier per unit dosage.
  • the dosage of the compounds according to this invention is 1-500 mg/day, e.g. about 100 mg per dose, when administered to patients, e.g. humans, as a drug.
  • a typical tablet that may be prepared by conventional tabletting techniques contains Core: Active compound (as free compound 100 mg or salt thereof)
  • the route of administration may be any route, which effectively transports the active compound to the appropriate or desired site of action, such as oral or parenteral e.g. rectal, transdermal, subcutaneous, intranasal, intramuscular, , topical, intravenous, intraurethral, ophthalmic solution or an ointment, the oral route being preferred.
  • oral or parenteral e.g. rectal, transdermal, subcutaneous, intranasal, intramuscular, , topical, intravenous, intraurethral, ophthalmic solution or an ointment, the oral route being preferred.
  • a mammalian cell line which can be obtained from the American Tissue Type Collection or other similar governmental or commercial sources, is , incubated with said modified compound. After incubation at conditions well known to those skilled in the art, the cells are washed appropriately, lysed and the lysate is isolated. Appropriate controls, well known to those skilled in the art, must be included.
  • a number of different procedures may in turn be used to extract and purify said compound from said lysate. Said compound may or may not retain the attached chemical group or said cyclic compound may or may not have been hydrolyzed.
  • a number of different procedures - well known to those skilled in the art may be used to structurally and chemically characterize said purified compound. Since said purified compound has been isolated from said cell lysate and hence has been taken up by said cell line, a comparison of said structurally and chemically characterized compound with that of the original unmodified compound (i.e.
  • said purified compound may be subjected to enzyme kinetic analysis as described in detail in the present invention. If the kinetic profile is similar to that of the original compound without said attached chemical group, but different from said modified compound, this confirms that said chemical group has been removed or said cyclic compounds has been hydrolyzed. Similar techniques may be used to analyze compounds of the invention in whole animals and mammals.
  • TLC thin layer chromatography
  • CDCI 3 deuterio chloroform
  • CD 3 OD tetradeuterio methanol
  • DMSO-d 6 hexadeuterio dimethylsulfoxide.
  • H NMR shifts ( ⁇ H ) are given in parts per million (ppm) down field from tetramethylsilane as internal reference standard.
  • M.p.: is melting point and is given in °C and is not corrected. Column chromatography was carried out using the technique described by W.C. Still et al., J. Org. Chem.
  • Example 2 The title compound was prepared in a similar way as described in Example 1 using 5-methyl-2-phenyl-2A-/-[1 ,2,3]triazole-4-carboxylic acid and 2-amino-5- aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material.
  • the hydrogenation step in Example 1 was omitted.
  • Example 2 The title compound was prepared in a similar way as described in Example 1 using 2-hydroxy-4-ethoxybenzoic acid and 2-amino-5-aminomethyl-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester as the starting material.
  • the hydrogenation step in Example 1 was omitted.
  • the title compound was prepared in a similar way as described in Example 1 using 2-hydroxy-4-ethoxybenzoic acid and 2-amino-5-aminomethyl-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester as the starting material.
  • the titie compound was prepared in a similar way as described in Example 1 using 5-methyI-2-phenyl-2H-[1 ,2,3]triazole-4-carboxylic acid and 2-amino- 5- aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material.
  • LC-MS: m/z: 485.2 [M+H] + HPLC (A1): R t 22.04 min
  • the titie compound was prepared in a similar way as described in Example 1 using 1H-indole-2-carboxylic acid and 2-amino-5-aminomethyl-6-(4-ethoxy- benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the titie compound was prepared in a similar way as described in Example 1 using 3-(biphenyl-4-yl)acrylic acid and 2-amino-5-aminomethyl-6-(4-methoxy- benzyI)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the titie compound was prepared in a similar way as described in Example 1 using 1-naphthyI-carboxylic acid and 2-amino-5-aminomethyl-6-(4-ethoxy- benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the hydrogenation step in Example 1 was omitted.
  • the cooled reaction mixture was concentration in vacuo and the residual oil was purified by silica gel chromatography using a mixture of hexanes/ethyl acetate (6:1) as eluent affording 474 mg (77 %) of a 1:1 mixture of 2-amino-7- (benzyloxycarbonylamino-methyl)-4,7-dihydro-5W-thieno[2,3-c]pyridine-3,6- dicarboxylic acid di-ferf-butyl ester and 2-amino-5-(benzyloxy-carbonylamino- methyl)-4,7-dihydro-5AV-thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester as a solids.
  • Salicylic acid (1.25 g, 9.05 mmol) was dissolved in anhydrous N,N-dimethyl- formamide (15 ml) and placed under nitrogen atmosphere.
  • Sodium hydride (0.76 g, 18.1 mmol) was added and the reaction stirred for 10 minutes, then benzyl bromide (4.3 ml, 36.2 mmol) was added.
  • the reaction was stirred at ambient temperature for 4 days.
  • the reaction mixture was diluted with ethyl acetate (75 ml) and washed with water (3 x 25 ml) followed by brine (15 ml).
  • the organic layer was dried (MgSO ), filtered, and the solvent evaporated in vacuo.
  • the residue was purified by silica gel chromatography affording 2.61 g (90 %) of 2- benzyloxy-benzoic acid benzyl ester as an oil.
  • the mixture was stirred overnight at room temperature.
  • the layers wee separated and the aqueous phase extracted with dichloromethane.
  • the combined organic phases were filtered through a plug of silica eluting with dichloromethane.
  • the relevant fractions were concentrated in vacuo.
  • the residue was dissolved in hot heptane and cooled. This leaves a yellowish gummy material on the side of the flask and crystals starts forming.
  • the heptane solution was heated again to dissolve crystals, leaving the gummy material on the side of the flask and the mixture was filtered hot.
  • the band consisting of a mixture of 5- and 7-isomer was collected and purified on a reverse phase (C 8 ) column using a Flash 40 system. The residue was applied in a minimum volume of acetonitrile and eluted with a mixture of 40% acetonitrile in water containing 0.1 % trifluoroacetic acid. When the first isomer (the 5-isomer) was collected the eluent was changed to 50% acetonitrile in water with 0.1% trifluoroacetic acid and the 7-isomer was collected.
  • the ⁇ -methyl benzyl protected amine was dissolved in a mixture of methanol/formic acid (9:1) (app. 10 ml/100 mg ⁇ -methyl benzyl protected amine). The solution was degassed by purging with nitrogen for 1 minute before 10% Pd/C (50% H 2 O content) was added. The reaction mixture was stirred at room temperature until TLC showed that all the starting material was consumed (typical 1-4 days). The Pd/C was filtered off using Celite and the filter cage was washed with plenty of methanol. The filtrate was concentrated in vacuo and the residue partitioned between ethyl acetate and water. The organic phase was separated, dried (MgSO 4 ) and filtered. The solvent was removed in vacuo and the crude product was purified using column chromatography (SiO 2 , and a mixture of ethyl acetate/methanol as eluent).
  • the ⁇ -methyl benzyl protected amine was dissolved in a mixture of isopropyl alcohol/formic acid (9:1) (app. 10 ml/100 mg ⁇ -methyl benzyl protected amine). The solution was degassed by purging with nitrogen for 1 minute before 10% Pd/C (dry) was added. The reaction mixture was stirred at room temperature until TLC showed that all the starting material was consumed (typical 1-4 days). The Pd/C was filtered off using Celite and the filter cage was washed with plenty of methanol. The filtrate was concentrated in vacuo and partitioned between ethyl acetate and water. The organic phase was separated, dried (MgSO 4 ) and filtered. The solvent was removed in vacuo and the crude product was purified using column chromatography (SiO 2 , and a mixture of ethyl acetate/methanol as eluent)
  • the carboxylic acid ferf-butyl ester was dissolved in a mixture of trifluoroacetic acid/dichloromethane (1 :1) (app. 1 ml/100 mg carboxylic acid ferf-butyl ester).
  • the reaction mixture was stirred for 16 hours at room temperature before diethyl ether (2 x the reaction volume) was added drop-wise.
  • the precipitate was filtered off/spun down on a centrifuge and washed with diethyl ether to give analytical pure titie compound.
  • thienor2,3-clpyridine-3-carboxylic acid The titie compound was prepared using 3-phenoxy-benzoic acid and 2-amino-5- (S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
  • the title compound was prepared using 4-benzoylamino-benzoic acid and 2- amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A,
  • the title compound was prepared using 4-benzoylamino-benzoic acid and 2- amino-5-(R)-aminomethyl-6-(1-(S)-phenyI-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
  • the titie compound was prepared using 3-hydroxy-7-methoxy-naphthalene-2- carboxylic acid and 2-amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above. Calculated for C 22 H ⁇ 9 N 3 O 7 S, 1xH 2 O, 0.7xC 2 HF 3 O 2 C, 49.06%; H, 4.00%; N, 7.03%. Found: C, 49.30%; H, 4.37%; N, 6.66%
  • the titie compound was obtained using the above ureido-compound and Method
  • the title compound was prepared using 4-acetylamino-benzenesulfonyI chloride and 2-amino-5-( ?)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. Formation of the sulpfonamide was performed in pyridine at room temperature using a slight excess of the sulfonyl chloride followed by Method B, C and D as described above.
  • the titie compound was prepared using 4-acetylamino-benzenesulfonyl chloride and 2-amino-5-(S)-aminomethyl-6-(1 -(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the title compound was prepared in a similar way as described in Example 1 using 4-benzyl-benzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the titie compound was prepared in a similar way as described in Example 1 using 5-ferf-butoxycarbonylamino-5-methyl-hex-2-enoic acid (prepared as described by Hansen et al. in J. Med. Chem. 41; (1998); 3705-3714) and 2- amino-5-aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
  • the titie compound was prepared from 2-(4,4-diethoxy-1-methylpiperidin-2- yImethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and method A, B, C and D in "General chiral synthesis"
  • the titie compound was prepared from 2-(4,4-diethoxy-1-methylpiperidin-2- ylmethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and method A, B, C and D in "General chiral synthesis"
  • 1 H-NMR 400 MHz, DMSO-d 6 ): ⁇ 13.78 (bs, 1H), 12.34 (bs, 1 H), 11.67 (s, 1H), 10.33 (bs, 1H), 8.79 (bs, 1 H, -CONWCH 2 ), 7.64 (d, 1 H), 7.44 (d, 1 H), 7.18 (m, 2H), 7.04 (m, 1 H), 4.46 (dd, 2H), 3.84-3.71 (m, 3H), 3.53 (bs, 1 H), 3.28 (d, 1H), 3.03 (m, 3H).

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Abstract

The present invention provides novel thienopyridines, novel compositions, methods of their use, and methods of their manufacture, where such compounds of Formula (I) are pharmacologically useful inhibitors of Protein Tyrosine Phosphatases (PTPase's) including PTP1B, T cell PTP (TC-PTP), wherein X, R1, R2, R3, and R4 are defined more fully in the description. The compounds are useful in the treatment of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance, obesity, and other diseases.

Description

Modulators of Protein Tyrosine Phosphatases (PTPases)
FIELD OF THE INVENTION
The present invention relates to novel thienopyridines, to methods for their preparation, to compositions comprising the compounds, to the use of these compounds as medicaments and their use in therapy, where such compounds of Formula 1 are pharmacologically useful inhibitors or modulators of Protein Tyrosine Phosphatases (PTPases) including PTP1 B and T cell PTP (TC-PTP),
Figure imgf000002_0001
Formula 1
wherein X, Ri, R2, R3, and R are defined more fully below.
It has been found that PTPases play a major role in the modulation and regulation of fundamental cellular signaling mechanisms involved in metabolism, growth, proliferation and differentiation (Fischer et al, Science 253:401-6 (1991); Tonks and Neel, Cell 87: 365-368 (1996); Neel and Tonks, Current Opinion in Cell Biology 9: 193-204 (1997); Hunter, Phil. Trans. R. Soc. Lond. B 353: 583- 605 (1998); Zhang, Critical Reviews in Biochemistry and Molecular Biology 33:1- 52 (1998)). There is increasing evidence which suggests that inhibition of these PTPases may help treat or manage certain types of diseases such as type 1 and type 2 diabetes, obesity, autoimmune diseases, acute and chronic inflammation, osteoporosis and various forms of cancer. In addition, certain infectious diseases may also be treated or managed by administration PTPase inhibitors (Clemens et al. , Molecular Microbiology 5: 2617-2620 (1991 )).
BACKGROUND OF THE INVENTION
Protein phosphorylation is now well recognized as an important mechanism utilized by cells to transduce and regulate signals during different stages of cellular function (Hunter, vide supra; Chan et al., Annu. Rev. Immunol. 12: 555- 592 (1994); Zhang, Curr. Top. Cell. Reg. 35: 21-68 (1997); Matozaki and Kasuga, Cell. Signal. 8: 113-19 (1996); Fischer et al, vide supra). The level of tyrosine phosphorylation is balanced by the opposing action of protein tyrosine kinases and protein tyrosine phosphatases (PTPases). There are at least two major classes of phosphatases: (1) those that dephosphorylate proteins (or peptides) that contain a phosphate group(s) on a serine or threonine moiety (termed Ser/Thr phosphatases) and (2) those that remove a phosphate group(s) from the amino acid tyrosine (termed protein tyrosine phosphatases or PTPases or PTPs). The PTPases are a family of enzymes that can be classified into two groups: a) intracellular or nontransmembrane PTPases and b) receptor-type or transmembrane PTPases. In addition, dual-specificity phosphatases and low molecular weight phosphatases can also dephosphorylate phosphotyrosyl proteins (WO97/39748, WO97/40017, WO99/15529, WO97/08934, WO98/27065, WO99/46236, WO99/46244, WO99/46267, WO99/46268, WO99/46237). It has been found that PTPases play a major role in the above modulation and regulation of fundamental cellular signaling mechanisms involved in metabolism, growth, proliferation and differentiation (Fischer et al, Science 253:401-6 (1991); Tonks and Neel, Ce// 87: 365-368 (1996); Neel and Tonks, Current Opinion in Cell Biology 9: 193-204 (1997); Hunter, Phil. Trans. R. Soc. Lond. B 353: 583-605 (1998); Zhang, Critical Reviews in Biochemistry and
Molecular Biology 33:1-52 (1998)). Reports from many laboratories have shown that PTPases can act both as positive and negative regulators of signal transduction processes. PTPases have been implicated in a variety of human diseases, including type 1 and type 2 diabetes, obesity, autoimmune diseases, acute and chronic inflammation, osteoporosis, proliferative disorders including various forms of cancer, growth disorders, and defective platelet aggregation (WO97/39748, WO97/40017, WO99/15529, WO97/08934, WO98/27065, WO99/46236, WO99/46244, WO99/46267, WO99/46268, WO99/46237). Accordingly, there is increasing evidence which suggests that inhibition of these PTPases may help treat or manage said diseases (Hunter, vide supra; Neel and Tonks, vide supra; Frangione et al., EMBO J. 12: 4843-4856; Zhang, Curr. Top. Cell. Reg. 35: 21-68 (1997); Zhang, vide supra; Evans and Jallan, Exp. Opinion. Invest. Drugs 8: 139-160 (1999); Burke and Zhang, Biopolymers (Peptide Science) 47: 225-241 (1998); Elchebly et al., Science 283: 1544-1548 (1999); Wrobel et al., J. Med. Chem. 42: 3199-3202 (1999)). In addition, certain infectious diseases may also be treated or managed by administration PTPase inhibitors (Clemens et al., Molecular Microbiology 5: 2617-2620 (1991)).
Both selective PTPase inhibitors and inhibitors that bind to several PTPases (non-selective inhibitors) can be used therapeutically to partially or completely restore PTPase-mediated perturbed signal transduction processes and thus for management, treatment or prevention of the above diseases.
WO 99/46267 discloses compounds, which are pharmacologically useful inhibitors of PTPases. However, the present invention, which represents a novel selection under WO 99/46267, discloses a class of compounds, which surprisingly are more potent against protein tyrosine phosphatases (e.g. PTP1B) than those disclosed in WO 99/46267.
DESCRIPTION OF THE INVENTION
The present invention relates to compounds of the Formula 1 wherein X, R^ R2, R3, and R are defined below;
Figure imgf000004_0001
Formula 1 wherein
X is -C(O)- or -S(O)2-; R! and R2 are independently hydrogen, C C6alkyl, aryl-R5-, R6-C(O)-O-R7- or aryl-R8-C(O)-O-R9- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, C Cδalkyl or C C6alkyloxy; R3 is hydrogen, C Cealkyl, H2N-R37-, C2-C6alkenyl, C2-C6alkynyl, aryl, aryl-Rι0-, aryl-N(R35)-, aryl-R11-N(R36)- , C C6alkyloxy or aryl-R13-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, d-Cealkyl, R38-aryl-, aryl, aryl- R14-, d-Cealkyloxy, aryloxy, aryl-R15-O-, aryl-N(R16)-, R18-C(O)-N(Rι9)-, aryl-
C(O)-N(R21)- or aryl-R23-C(O)-N(R24)- and wherein aryl is phenyl, naphthyl or thiophenyl;
R4 is hydrogen, C C6alkyl, aryl-R26-, R27-O-C(O)-, aryl-R28-O-C(O)-, R29-C(O)-O- R30-O-C(O)- or aryl-R3 C(O)-O-R32-O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, aryl, aryl-R33-, C C6aIkyloxy or aryl-R34-O- and wherein aryl group is phenyl, naphthyl or thiophenyl; and wherein R5, R7, R8, Rg, R10, Rn, Rι3> Rι4, R15. R23> R2β. R28ι 30. R3i> R32. R331 R3 ,and Rs7 independently are C1-6-alkylene, wherein R6, Rι2, R17, Rιs, R20, R22,
R25, R27 and R29 independently are C1-6alkyl and wherein R 6, Rι9, R2ι, R24, R35,
R36 and R38 independently are hydrogen or C1-6alkyl; 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 f tautomeric form.
The compounds of the invention can be further modified to act as prodrugs.
A preferred prodrug is acetoxymethyl esters or acetoxymethyl carbamates of the compounds of the present invention. As a general procedure preparation of an acetoxymethyl ester is given below (C.Schultz et al, The Journal of Biological Chemistry, 1993, 268, 6316-6322.;.-
A carboxylic acid (1 equivalent) is suspended in dry acetonitrile (2 ml per 0.1 mmol). Diisopropyl amine (3.0 equivalents) is added followed by bromomethyl acetate (1.5 equivalents). The mixture is stirred under nitrogen overnight at room temperature. Acetonitrile is removed under reduced pressure to yield an oil which is diluted in ethyl acetate and washed with water (3 x). The organic layer is dried over anhydrous magnesium sulfate, filtred and the solvent removal under reduced pressure affording a crude oil. The product is purified by column chromatography on silica gel, using an appropriate solvent system known to those skilled in the arte. DEFINITIONS
As used herein, the term "attached" or "-" signifies a stable covalent bond, certain preferred points of attachment points being apparent to those skilled in the art.
The terms "halogen" and "halo" includes fluorine, chlorine, bromine, and iodine.
The term "alkyl" includes C^Ce straight chain saturated, C C6 branched chain saturated and C3-C6 cyclic saturated hydrocarbon groups. For example, this definition shall include but is not limited to methyl (Me), ethyl (Et), propyl (Pr), butyl (Bu), pentyl, hexyl, isopropyl (i-Pr), isobutyl (i-Bu), fe/f-butyl (t- u), sec-butyl
(s-Bu), cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.
The term "alkenyl" includes C2-C6 unsaturated aliphatic hydrocarbon groups and
C2-C6 branched unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms and at lest one double bond. For example, this definition shall include but is not limited to ethenyl, propenyl, butenyl, pentenyl, hexenyl, isopentenyl, neopentenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, and the like.
The term "alkynyl" includes C2-C6 straight chain unsaturated aliphatic, C2-C6 branched unsaturated and cyclic C6 unsaturated aliphatic hydrocarbon groups having the specified number of carbon atoms and at lest one triple bond. For example, this definition shall include but is not limited to acetynyl, propynyl, butynyl, pentynyl, hexynyl, cyclohexynyl and the like.
The term "alkyloxy" (e.g. methoxy, ethoxy, propyloxy, allyloxy, cyclohexyloxy) represents an "alkyl" group as defined above having the indicated number of carbon atoms attached through an oxygen bridge.
The term "aryloxy" (e.g. phenoxy, naphthyloxy and the like) represents an aryl group as defined below attached through an oxygen bridge.
The term "aryl" represents an unsubstituted, mono-, di- or trisubstituted monocyclic, polycyclic, biaryl or heterocyclic aromatic group(s) covalently attached at any ring position capable of forming a stable covalent bond, certain preferred points of attachment being apparent to those skilled in the art (e.g., 3- indolyl, 4(5)-imidazolyl).
The definition of aryl includes phenyl, biphenyl, indenyl, naphthyl (1 -naphthyl, 2- naphthyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1 ,2,3-triazoM-yl, 1 ,2,3-triazol-2-yl 1 ,2,3-triazol-4-yl, 1 ,2,4-triazol-3-yl), thiophenyl (2-thiophenyl, 3-thiophenyl, 4-thiophenyl, 5-thiophenyl), pyridyl (2- pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1 -isoquinolyl, 3- isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5- benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl).
It is a well known problem in drug discovery that compounds, such as enzyme inhibitors, may be very potent and selective in biochemical assays, yet be, inactive in vivo. This lack of so-called bioavailability may be ascribed to a number of different factors such as lack of or poor absorption in the gut, first pass metabolism in the liver, poor uptake in cells. Although the factors determining bioavailability are not completely understood, there are many examples in the ., scientific literature - well known to those skilled in the art - of how to modify compounds, which are potent and selective in biochemical assays but show low , or no activity in vivo, into drugs that are biologically active. By the term Original compound' is understood a compound of Formula 1 wherein R-i and R2 are both hydrogen. It is within the scope of the invention to modify the original corηpounds of the invention by attaching chemical groups that will improve the bioavailability of said compounds in such a way that the uptake in cells or mammals is facilitated. Examples of said modifications, which are not intended in any way to limit the scope of the invention, include changing of one or more of the carboxy groups at the R and R2 position to esters (for instance methyl esters, ethyl esters, acetoxymethyl esters or other acyloxymethyl esters). Original compounds of the invention modified by attaching chemical groups are termed 'modified compounds'. Other examples of modified compounds, which are not intended in any way to limit the scope of the invention, are compounds that have been, cyclized at specific positions - so called 'cyclic compounds' - which upon uptake in cells or mammals become hydrolyzed at the same specific position(s) in the molecule to yield the compounds of the invention, the original compounds, which are then said to be 'non-cyclic'. For the avoidance of doubt, it is understood that the latter original compounds in most cases will contain other cyclic or heterocyclic structures that will not be hydrolyzed after uptake in cells or mammals. Generally, said modified compounds may not show behavior in biochemical assays similar to that of the original compound, i.e. the corresponding compounds of the invention without the attached chemical groups or said modifications. Said modified compounds may even be inactive in biochemical assays. However, after uptake in cells or mammals these attached chemical groups of the modified compounds may in turn be removed spontaneously or by endogenous enzymes or enzyme systems to yield compounds of the invention, original compounds. 'Uptake' is defined as any process that will lead to a substantial concentration of the compound inside cells or in mammals. After uptake in cells or mammals and after removal of said attached chemical group or hydrolysis of said cyclic compound, the compounds may have the same structure as the original compounds and thereby regain their activity and hence become active in cells and/or in vivo after uptake. Thus, the term 'a functional group which can be converted to hydrogen in vivo' is intended to include any group which upon administering the present compounds to the subjects in need thereof can be converted to hydrogen e.g. enzymatically or by the acidic environment in the stomach.
The compounds of the present invention have asymmetric centres and may occur as racemates, racemic mixtures, and as individual enantiomers or diastereoisomers, with all isomeric forms being included in the present invention as well as mixtures thereof.
Pharmaceutically acceptable salts of the compounds of Formula 1 , where a basic or acidic group is present in the structure, are also included within the scope of this invention. When an acidic substituent is present, such as -COOH, 5- tetrazolyl or -P(O)(OH)2ι there can be formed the ammonium, morpholinium, sodium, potassium, barium, calcium salt, and the like, for use as the dosage form. When a basic group is present, such as amino or a basic heteroaryl radical, such as pyridyl, an acidic salt, such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroacetate, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartarate, fumarate, mandelate, benzoate, cinnamate, ethanesulfonate, ethane sulfonate, picrate and the like, and include acids related to the pharmaceutically acceptable salts listed in Journal of Pharmaceutical Science, 66, 2 (1977) and incorporated herein by reference, can be used as the dosage form. Also, in the case of the -COOH or -P(O)(OH)2 being present, pharmaceutically acceptable esters can be employed, e.g., methyl, tert-butyl, pivaloyloxymethyl, and the like, and those esters known in the art for modifying solubility or hydrolysis characteristics for use as sustained release or prodrug formulations. In addition, some of the compounds of the present invention may form solvates with water or common organic solvents. Such solvates are encompassed within the scope of the invention.
The term "therapeutically effective amount" shall mean that amount of drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor or other.
PREFERRED EMBODIMENTS OF THE INVENTION
In a preferred embodiment, the present invention is concerned with compounds of Formula 1
Figure imgf000009_0001
Formula 1 wherein X is -C(O)-;
Ri and R2 are independently hydrogen, d-C6alkyl, aryl-R5-, R6-C(O)-O-R7- or aryl-R8-C(O)-O-R9- wherein aryl is phenyl, naphthyl, thiophenyl, which aryl group is optionally substituted with halogen, trihalomethyl, aryl, aryl- , Rio-, C C6alkyloxy or aryl-Rι O-; R3 is CrC6alkyl, H2N-R35-, C2-C6alkenyl, C2-C6alkynyl, aryl-Rι2-, aryl, aryl-R13-, aryl-N(Rι4)-, aryl-R 5-N(R16)-, R38-aryl-, C C6alkyloxy or aryl-Rι7-O- wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, or indolyl, which aryl group is optionally substituted with halogen, hydroxy, d-C6alkyl, aryl, aryl-Rι8-, CrC6alkyloxy, aryloxy, aryl-R19-O-; R4 is hydrogen, d-Cealkyl, aryl-R20-, R2rC(O)-O-R22-O-C(O)- or aryl-R23-C(O)-O- R24-O-C(O)- wherein aryl is phenyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, aryl, aryl-R25-, d-C6alkyloxy or aryl-R26-O- and wherein aryl group is phenyl, naphthyl or thiophenyl; and wherein R5, R7, R8, R9, R10, Rn, Rι3, R15, R171 Rie. i9, R2o, R22, R23, R24, R25, R26, and R35 independently are d-Cβ-alkylene, wherein Rι2 is alkenylene, wherein R6, R2ι, and R38 independently are CrC6alkyl and wherein R14 and R16 independently are hydrogen or Cι-6alkyl;
More preferred compounds of the invention are compounds of Formula 1
Figure imgf000010_0001
wherein
X is -C(O)-;
Ri and R2 are independently hydrogen or C C6alkyl;
R3 is CrC6alkyl, H2N-R18, aryl-R5-, aryl, aryl-R6-, aryl-N(R7)-, aryl-R8-O- or R19- aryl- wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, or indolyl, which aryl group is optionally substituted with halogen, hydroxy, CrCealkyl, aryl, aryl-R9-, C C6alkyloxy, aryloxy, aryl-R10-O-; R4 is hydrogen, aryl-Rn-, Rι2-C(O)-O-R13-O-C(O)- or aryl-Rι -C(O)-O-R15-O- C(O)- wherein aryl is phenyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, aryl, aryl-Rι6-, C C6alkyloxy or aryl-Ri7-O- and wherein aryl group is phenyl, naphthyl or thiophenyl; and wherein R5, R8, R9, R10, Rn, Rι3, Rι , Rι5, ie, 17 and Rι8 independently are Cr C6-alkylene, wherein R6 is alkenylene, wherein R7 and Rι2 independently are C C6alkyl and wherein R7 and Ri9 independently are hydrogen or d-C6alkyl;
Even more preferred compounds of the invention are compounds of Formula 1
Figure imgf000010_0002
wherein X is -C(O)-;
Ri, R2 and R* are hydrogen;
R3 is CrC6alkyl, H2N-Rι2-, aryl, aryl-R5-, aryl-N(R6)-, aryl-R7-O- or R13-aryl-, wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, or indolyl, which aryl group is optionally substituted with halogen, hydroxy, d-Cealkyl, aryl, aryl-R8-, CrC6alkyloxy, aryloxy, aryl-R9-O- wherein aryl is phenyl or thiophenyl, which aryl group is optionally substituted with halogen, hydroxy, nitro, cyano, trihalomethyl, aryl, aryl-R10-, CrC6alkyloxy aryloxy, or aryl-Rn-O- and wherein aryl group is phenyl, naphthyl or thiophenyl; wherein R5, R7, R8, R9, Rι0, Rn and Rι2 independently are d-C6-alkylene and wherein R6 and Rι3 independently are hydrogen or Cι-6alkyl;
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 form.
The following compounds are preferred:
5-(Benzoylamino-methyl)-2-(oxaIyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c] pyridine-3-carboxylic acid;
5-(((6-Bromo-2-p-tolyl-quinoline-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-
2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(((5-Benzyloxy-1H-indole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-
(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-((4-Benzoylamino-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-5-(((5-methyl-2-phenyl-2H-[1 ,2,3]triazole-4- carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-
3-carboxylic acid;
5-((4-Ethoxy-2-hydroxy-benzoylamino)-methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(((Biphenyl-4-carbonyl)-amino)-methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(((Biphenyl-4-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Ethoxy-2-hydroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(((5-Methyl-2-phenyl-2H-[1 ,2,3]triazole-4-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-((3-Biphenyl-4-yl-acryloylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-(((1H-lndole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-(((1H-lndole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((3-Biphenyl-4-yl-propionylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid; 6-(4-Methoxy-benzyl)-5-(((naphthalene-1-carbonyl)-amino)-methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(Benzyloxycarbonylamino-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
7-(benzyloxycarbonylamino-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((2-Hydroxy-benzoylamino)-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((3-Hydroxy-benzoylamino)-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c] pyridine-3-carboxylic acid; 2-(Oxalyl-amino)-5-(S)-((3-phenoxy-benzoylamino)methyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-(R)-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((1-Hydroxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((3-Hydroxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(S)-(((3-Hydroxy-7-methoxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 2-(Oxalyl-amino)-5-(S)-((3-phenoxy-benzoylamino)methyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-acetoxymethyl ester;
2-(Oxalyl-amino)-5-(S)-(3-(4-phenoxy-phenyl)ureidomethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridin-3-carboxylic acid; 5-((4-Benzyl-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((5-Amino-5-methyl-hexanoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((Biphenyl-4-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-((4-Benzyl-benzoylamino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((Naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]-pyridine-3-carboxylic acid;
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 6-Benzyl-5-(S)-(((naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
6-Benzyl-5-(S)-(((1 - -indole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
or a pharmaceutically acceptable salt thereof.
In another embodiment the invention relates to a compound of Formula 1
Formula 1
wherein X is -C(O)- or -S(O)2-;
Ri and R2 are independently hydrogen or a functional group that can be converted to hydrogen in vivo;
R3 is hydrogen, CrC6alkyl, H2N-R35-, C2-C6alkenyl, C2-C6alkynyl, aryl, aryl-Rι0-, aryl-N(R35)-, aryl-Rn-N(R36)- , d-C6alkyloxy or aryl-R13-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R38- R14-, CrCealkyloxy, R38-O-, R38-Rι5-O-, R38-N(R16)-, R18-C(O)-N(Rι9)-, R38-C(O)- N(R2ι)- or R38-R23-C(O)-N(R24)-;
R4 is hydrogen, CrC6alkyl, aryl-R26-, R27-O-C(O)-, aryl-R28-O-C(O)-, R29-C(O)-O- R30-O-C(O)- or aryl-R3rC(O)-O-R32-O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, C C6alkyloxy or R39-R34-O-; and wherein R5, R7, R8, R9, R 0, Rn, R13, Rι4, R15, R23. R26, R28» R30, R31, R32, R33, R34 and R35 independently are CrC6alkylene, wherein R6, Rι2, R17, Rι8, R2o, R22, R25, 27 and R29 independently are CrC6alkyl and wherein Rι6, R19, R2ι, R24, R35 and R36, and R37 independently are hydrogen or CrC6alkyl; R38 and R39 are independently phenyl, naphthyl or thiophenyl
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 form.
In another embodiment R and R2 are independently hydrogen, Cι-C6alkyl, aryl- R5-, R6-C(O)-O-R7- or aryl-R8-C(O)-O-R9- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, CrC6alkyl or d-C6alkyloxy;
In another embodiment X is C(O).
In another embodiment X is S(O)2.
In another embodiment R1 and R2 are independently hydrogen, CrC6alkyl, aryl- R -, or R6-C(O)-O-R7-, wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, d-C6alkyl or Ci- Cealkyloxy.
In another embodiment Ri and R2 are independently hydrogen or d-Cealkyl.
In another embodiment Ri and R2 are hydrogen.
In another embodiment R3 is C C6alkyl, C2-C6alkenyl, C2-C6alkynyl, aryl, aryl- R10-, CrC6alkyloxy, H2N-R35- or aryl-Rι3-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R38- Ri4-, CrCealkyloxy, Ras-O-, R38-Rι5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)- N(R2 - or R38-R23-C(O)-N(R24)-.
In another embodiment R3 is CrC6alkyl, C2-C6alkenyl, aryl, H2N-R35- or aryl-R13- O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl , group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrCealkyl, R37-aryl-, R38-, R38-Ri4-, d-C6alkyloxy, R38-O-, R38-Rι5-O-, Rs8-N(Rι6)- , Rι8-C(O)-N(Rι9)-, R38-C(O)-N(R2i)- or R38-R23-C(O)-N(R24)-.
In another embodiment R3 is d-C6alkyl or aryl, wherein aryl is phenyl, biphenyl, . indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R38- Rι4-, CrCealkyloxy, R38-O-, R38-Rι5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)- N(R2ι)- or R38-R23-C(O)-N(R24)-.
In another embodiment R3 is aryl, wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R^-R^-, Ci- Cealkyloxy, R38-O-, R38-Ri5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)-N(R2ι)- or R38-R23-C(O)-N(R24)-. In another embodiment the aryl of R3 is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, quinolyl or indolyl.
In another embodiment the aryl of R3 is phenyl, naphthyl, or indolyl.
In another embodiment the aryl of R3 is phenyl.
In another embodiment the aryl of R3 is substituted by halogen, hydroxy, d- C6alkyl, R37-aryl-, R38-, R38-Rι -, CrC6alkyloxy, R38-O-, R38-Rι5-O-, Rι8-C(O)- N(Rι9)-, or R38-C(O)-N(R2ι)-.
In another embodiment the aryl of R3 is substituted by hydroxy, CrC6alkyl, R38-, R38-Rι -, CrCealkyloxy, R38-O-, Rι8-C(O)-N(Rι9)-, or R38-C(O)-N(R2ι)-.
In another embodiment R38 is phenyl or thiophenyl.
In another embodiment R4 is hydrogen, CrC6alkyl, aryl-R26-, R29-C(O)-O-R30-O-, C(O)- or aryl-R3rC(O)-O-R32-O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, CrCealkyloxy or R39-R34-O-.
In another embodiment R4 is hydrogen, CrC6alkyl, or aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, CrC6alkyloxy or R39-R34-O-.
In another embodiment R is hydrogen or aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, CrC6alkyloxy or R39-R34-O-.
In another embodiment R4 is hydrogen.
In another embodiment R4 is aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, d-C6alkyloxy or R39-R3 -O-.
In another embodiment the aryl of R4 is phenyl. In another embodiment the aryl of R4 is substituted by R^-.
In another embodiment R39 is CrC6alkyloxy.
In another embodiment R39 is methoxy.
Another aspect of the invention is compounds according to the invention that act as inhibitors of Protein Tyrosine Phosphatases.
Another aspect of the invention is a pharmaceutical composition comprising a compound of the invention or a pharmaceutically acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents.
Another aspect of the invention is a pharmaceutical composition suitable for treating type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity comprising a compound of the invention or a pharmaceutically acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents.
Another aspect of the invention is a pharmaceutical composition suitable for treating immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases comprising a compound of the invention or a pharmaceutical acceptable 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 form together with one or more pharmaceutically acceptable carriers or diluents. The composition may be in the form of an oral dosage unit or parenteral dosage unit.
The pharmaceutical composition may said may contain a compound of the invention such that it is administered as a dose in a range from about 0.05 to 1000 mg, preferably from about 0.1 to 500 mg and especially in the range from 50 to 200 mg per day
Another aspect of the invention is a compound of the invention or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable aqid or base, or any optical isomer or mixture of optical isomers, including a racemic mixture, or any tautomeric form for therapeutical use in the treatment or , prevention of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity.
Another aspect of the invention is a compound of the invention or a pharmaceutically acceptable 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 form for therapeutical use in the treatment or preventing of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases.
Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form as a medicament.
Another aspect of the invention is the use of a compound of the invention for preparing a medicament. Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form for the preparation of a medicament suitable for the treatment or preventing of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity.
Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable 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 form for the preparation of a medicament suitable for the treatment or preventing of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases.
Another aspect of the invention is a method of treating type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity comprising administering to a subject in need thereof an effective amount of a compound of the invention.
Another aspect of the invention is a method of treating immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases comprising administering to a subject in need thereof an effective amount of a compound of the invention to said subject.
Another aspect of the invention is a process for the manufacture of a medicament, particular to be used in the treatment or prevention of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance or obesity which process comprising bringing a compound of the invention or a pharmaceutically acceptable salt thereof into a galenic dosage form.
Another aspect of the invention is a process for the manufacture of a medicament, particular to be used in the treatment or prevention of immune dysfunctions including autoimmunity, diseases with dysfunctions of the coagulation system, allergic diseases, osteoporosis, proliferative disorders including cancer and psoriasis, diseases with decreased or increased synthesis or effects of growth hormone, diseases with decreased or increased synthesis of hormones or cytokines that regulate the release of/or response to growth hormone, diseases of the brain including Alzheimer's disease and schizophrenia, and infectious diseases which process comprising bringing a compound of the invention or a pharmaceutically acceptable salt thereof into a galenic dosage form.
Another aspect of the invention is a method for preparing a compound of formula
1 , characterized in
A)
Figure imgf000020_0001
a) NC-CH2-COORι, sulphur, morpholine or triethylamine, and EtOH; b) R2-O- C(O)-C(O)-imidazol-1-yl, THF; c) 25% TFA/CH2CI2; wherein ,, R2, R3, R4 and X are defined in claim 1 ;
The reaction step a) in Method A gives a mixture of regioisomers which can be separated by use of column chromatography known to thus skilled in the art. B)
Figure imgf000021_0001
a) By allowing an activated carboxylic acid or sulfonic acid (I); wherein X is - C(CO)- and W is -OH, -OSO2Me, halogen, R4COO- or X is -SO2- and W is chloride, a substituted 5-aminomethyl-tetrahydro-thieno[2,3-c]pyridine (II) to react under conditions known to thus skilled in the art which favour amide or sulfon amide bond formation followed by b) R2-O-C(O)-C(O)-imidazol-1-yl, THF and c) 25% TFA/CH2CI2; to yield (III) wherein R R2, R3, and R4 are defined in claim 1 ;
PHARMACOLOGICAL METHODS
The compounds are evaluated for biological activity with a truncated form of PTP1 B (corresponding to the first 321 amino acids), which was expressed in E. coli and purified to apparent homogeneity using published procedures well- known to those skilled in the art. The enzyme reactions are carried out using standard conditions essentially as described by Burke et al. (Biochemistry 35; 15989-15996 (1996)). The assay conditions are as follows. Appropriate concentrations of the compounds of the invention are added to the reaction mixtures containing different concentrations of the substrate, p-nitrophenyl phosphate (range: 0.16 to 10 mM - final assay concentration). The buffer used was 100 mM sodium acetate pH 5.5, 50 mM sodium chloride, 0.1 % (w/v) bovine serum albumin and 5 mM dithiothreitol (total volume 100 ml). The reaction was started by addition of the enzyme and carried out in microtiter plates at 25 °C for 60 minutes. The reactions are stopped by addition of NaOH. The enzyme activity was determined by measurement of the absorbance at 405 nm with appropriate corrections for absorbance at 405 nm of the compounds and p-nitrophenyl phosphate. The data are analyzed using nonlinear regression fit to classical Michaelis Menten enzyme kinetic models. Inhibition is expressed as K| values in μM. The results of representative experiments are shown in Table 1. Table 1
Inhibition of classical PTP1B by compounds of the invention
Figure imgf000022_0002
THE SYNTHESIS OF THE COMPOUNDS In accordance with one aspect of the invention, the compounds of the invention are prepared as illustrated in the following reaction scheme:
Method A
Figure imgf000022_0001
a) NC-CH2-COORι, sulphur, morpholine or triethylamine, and ethanol; b) R2-O- C(O)-C(O)-imidazol-1-yl, tetrahydrofurane; c) 25% trifluoroacetic acid/dichloromethane; wherein Rn, R2, R3, R4 and X are defined above;
The reaction step a) in Method A gives a mixture of regioisomers which can be separated by use of column chromatography known to thus skilled in the art.
Method B
Figure imgf000023_0001
a) By allowing an activated carboxylic acid or sulfonic acid (I); wherein X is -
C(CO)- and W is -OH, -OSO2Me, halogen, R4COO- or X is -SO2- and W is chloride, a substituted 5-aminomethyl-tetrahydro-thieno[2,3-c]pyridine (II) to react under conditions known to thus skilled in the art which favour amide or sulfon amide bond formation followed by b) R2-O-C(O)-C(O)-imidazol-1-yl, tetrahydrofuran and c) 25% trifluoroacetic acid/dichloromethane; to yield (III) wherein R1 ( R2, R3, and R4 are defined above;
Pharmacological Preparations
For the above indications the dosage will vary depending on the compound of the invention employed, on the mode of administration and on the therapy desired. However, in general, satisfactory results are obtained with a dosage of from about 0.5 mg to about 1000 mg, preferably from about 1 mg to about 500 mg of compounds of the invention, conveniently given from 1 to 5 times daily, optionally in sustained release form. Usually, dosage forms suitable for oral administration comprise from about 0.5 mg to about 1000 mg, preferably , from about 1 mg to about 500 mg of the compounds of the invention admixed with a pharmaceutical carrier or diluent.
The compounds of the invention may be administered in a pharmaceutically acceptable acid addition salt form or where possible as a metal or a -β- alkylammonium salt. Such salt forms exhibit approximately the same order of activity as the free acid forms.
This invention also relates to pharmaceutical compositions comprising a compound of the invention or a pharmaceutically acceptable salt thereof and, usually, such compositions also contain a pharmaceutical carrier or diluent. The compositions containing the compounds of this invention may be prepared by conventional techniques and appear in conventional forms, for example capsules, tablets, solutions or suspensions.
The pharmaceutical carrier employed may be a conventional solid or liquid carrier. Examples of solid carriers are lactose, terra alba, sucrose, talc, gelatine, agar, pectin, acacia, magnesium stearate and stearic acid. Examples of liquid carriers are syrup, peanut oil, olive oil and water. Similarly, the carrier or diluent may include any time delay material known to the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. If a solid carrier for oral administration is used, the preparation can be tabletted, placed in a hard gelatine capsule in powder or pellet form or it can be in the form of a troche or lozenge. The amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
Generally, the compounds of this invention are dispensed in unit dosage form comprising 10-200 mg of active ingredient in or together with a pharmaceutically acceptable carrier per unit dosage.
The dosage of the compounds according to this invention is 1-500 mg/day, e.g. about 100 mg per dose, when administered to patients, e.g. humans, as a drug.
A typical tablet that may be prepared by conventional tabletting techniques contains Core: Active compound (as free compound 100 mg or salt thereof)
Colloidal silicon dioxide (Areosil®) 1.5 mg
Cellulose, microcryst. (Avicel®) 70 mg Modified cellulose gum (Ac-Di-Sol®) 7.5 mg Magnesium stearate
Coating:
HPMC approx. 9 mg *Mywacett® 9-40 T approx. 0.9 mg
*Acylated monoglyceride used as plasticiser for film coating.
The route of administration may be any route, which effectively transports the active compound to the appropriate or desired site of action, such as oral or parenteral e.g. rectal, transdermal, subcutaneous, intranasal, intramuscular, , topical, intravenous, intraurethral, ophthalmic solution or an ointment, the oral route being preferred.
A number of procedures, well known to those skilled in the art, may be used to verify that the attached chemical groups have been removed or that the cyclic compound has been hydrolyzed after uptake in cells or mammals. An example, ., which is not intended in any way to limit the scope of the invention, is given in the following. A mammalian cell line, which can be obtained from the American Tissue Type Collection or other similar governmental or commercial sources, is , incubated with said modified compound. After incubation at conditions well known to those skilled in the art, the cells are washed appropriately, lysed and the lysate is isolated. Appropriate controls, well known to those skilled in the art, must be included. A number of different procedures, well known to those skilled in the art, may in turn be used to extract and purify said compound from said lysate. Said compound may or may not retain the attached chemical group or said cyclic compound may or may not have been hydrolyzed. Similarly, a number of different procedures - well known to those skilled in the art - may be used to structurally and chemically characterize said purified compound. Since said purified compound has been isolated from said cell lysate and hence has been taken up by said cell line, a comparison of said structurally and chemically characterized compound with that of the original unmodified compound (i.e. without said attached chemical group or said non-cyclic compound) will immediately provide those skilled in the art information on whether the attached chemical group as been removed in the cell or if the cyclic compound has been hydrolyzed. As a further analysis, said purified compound may be subjected to enzyme kinetic analysis as described in detail in the present invention. If the kinetic profile is similar to that of the original compound without said attached chemical group, but different from said modified compound, this confirms that said chemical group has been removed or said cyclic compounds has been hydrolyzed. Similar techniques may be used to analyze compounds of the invention in whole animals and mammals.
EXAMPLES
The process for preparing compounds of Formula 1 and preparations containing them is further illustrated in the following examples, which, however, are not to be construed as limiting.
Hereinafter, TLC is thin layer chromatography, CDCI3 is deuterio chloroform, CD3OD is tetradeuterio methanol and DMSO-d6 is hexadeuterio dimethylsulfoxide. The structures of the compounds were confirmed by either elemental analysis or NMR, where peaks assigned to characteristic protons in the title compounds are presented where appropriate. H NMR shifts (δH) are given in parts per million (ppm) down field from tetramethylsilane as internal reference standard. M.p.: is melting point and is given in °C and is not corrected. Column chromatography was carried out using the technique described by W.C. Still et al., J. Org. Chem. 43: 2923 (1978) on Merck silica gel 60 (Art. 9385). HPLC analyses are performed using 5μm C18 4 x 250 mm column eluted with various mixtures of water and acetonitrile, flow = 1 ml/min, as described in the experimental section.
Compounds used as starting material are either known compounds or compounds, which can readily be prepared by methods known per se.
EXAMPLE 1
Figure imgf000027_0001
5-(Benzoylamino-methyl)-2-(oxalyl-amino)-4.5,6,7-tetrahvdro-thienor2.3-c1 pyridine-3-carboxylic acid
To a solution of 4-methoxybenzyl amine (22.63 ml, 0.173 mol), 4 A molecular sieves (18 g), and dichloromethane (400 ml) was added ethyl glyoxalate (38.86 ml (50 % solution in toluene), 0.19 mol). After stirring for 1 hr, the solution was cooled to 0 °C and trifluoroacetic acid (13.34 ml, 0.173 mol) was added via syringe. After 10 min, boron trifluoride etherate (21.94 ml, 0.173 mol) was added via syringe and the reaction mixture allowed to stir for 10 min. A solution of 2- trimethylsilyloxy-1 ,3-butadiene (25 g, 0.173 mol) in dichloromethane (30 ml) was added dropwise over a 5 min period. The reaction mixture was allowed to stir at 0 °C for 4 hours, at which time it was quenched with saturated sodium bicarbonate (100 ml). The mixture was filtered and the organic layer washed with saturated sodium bicarbonate (3 x 300 ml) and water (500 ml). The organic layer was dried (MgSO4), filtered and evaporated in vacuo. The residue (51 g) was subjected to flash chromatography using a mixture of hexanes/ethyl acetate (60:40) as eluent which afforded 16.5 g (33 %) of 4-oxo-1-(4-methoxy-benzyl)-piperidine-2- carboxylic acid ethyl ester as an oil. 1H-NMR (CDCI3) δ 7.28 (d, 2H, J = 8.4 Hz), 6.86 (d, 2H, J = 8.4 Hz), 4.20 (dq, 2H, J = 7.2 and J = 1.8 Hz), 3.97 (s, 3H), 3.70 (d, 2H, J = 3.6 Hz), 3.01 (ddd, 1 H„ J = 15.3, J = 8.7 and J = 5.4 Hz), 2.88-2.81 (m, 1 H), 2.56 (dd, 1 H, J = 9.9 and J = 6 Hz), 2.45-2.40 (m, 1H), 1.29 (t, 3H, J = 7.2 Hz); LC-MS; Rt=1.54, m/z: 292.3 [M+1]+.
To a solution of 4-oxo-1-(4-methoxy-benzyl)-piperidine-2-carboxylic acid ethyl ester (26.3 g, 0.09 mmol) in absolute ethanol (500 ml) was added tert- butylcyanoacetate (16.6 g, 0.12 mol), sulfur (3.18 g, 0.099 mol), and morpholine (15.7 ml, 0.18 mol). The reaction was placed in a 50 °C oil bath and stirred under nitrogen atmosphere for 18 hours. The mixture was cooled to ambient temperature, concentrated in vacuo. and the residue purified by silica gel chromatography using a gradient of ethyl acetate/hexane, (10 % to 20 % gradient) as eluent in two portions, which afforded a mixture of 2-amino-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3,5-dicarboxylic acid 3- fe/f-butyl ester 5-ethyl ester and 2-amino-6-(4-methoxy-benzyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3,7-dicarboxylic acid 3-ferf-butyI ester 7-ethyl ester 16.4 g (41 %).
1H-NMR (400 MHz, CDCI3) δ 7.28 (d, J = 8 Hz, 2H), 6.86 (d, J = 8 Hz, 2H), 6.00- 5.91 (2s, 2H), 4.24-4.13 (m, 2H), 3.83-2.64 (m, 7H), 3.80 (s, 3H), 1.54-1.52 (2s, 9H) LC-MS: Rt = 3.01 min, m/z: 447.2 [M+H]+
A mixture of 2-amino-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3,5-dicarboxylic acid 3-ferf-butyl ester 5-ethyl ester and 2-amino-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3,7-dicarboxylic acid 3- ferf-butyl ester 7-ethyl ester (16.4 g, 36.7 mmol) was dissolved in anhydrous tetrahydrofuran (250 ml), treated with lithium borohydride (3.99 g, 184 mmol) and heated in a oil bath at 45 °C for 16 hours. TLC analysis showed that the reaction was incomplete. Additional lithium borohydride (0.8 g, 36.7 mmol) was added and the reaction stirred at 50 °C for an additional 6 hours. The solution was cooled to ambient temperature and neutralized by addition of 1 N hydrochloric acid until no bubbling was observed and final pH = 7 was obtained. The precipitate was filtratred off and the filtrate extracted with ethyl acetate (3 x 100 ml). The combined organic phases were washed with brine (100 ml), dried (MgSO4), and the solvent evaporated in vacuo affording 14.4 g of a crude , mixture of 2-amino-5-hydroxymethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid tert-buty\ ester and 2-amino-7- hydroxymethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid terf-butyl ester as a solid.
1H-NMR (400 MHz, CDCI3) δ 7.23-7.19 (m, 2H), 6.87-6.82 (m, 2H), 5.99-5.91 (2s, 2H), 3.80 (s, 3H), 3.79-2.40 (m, 9H), 1.56-1.52 (2s, 9H) LC-MS: Rt = 2.50 min, m/z: 405.4 [M+H]+
A solution of 2-amino-5-hydroxymethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester and 2-amino-7- hydroxymethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester (14.4 g, 35.6 mmol), phthalimide (5.76 g, 39.2 mmol), and triphenyl phosphine (10.3 g, 39.2 mmol) was prepared in anhydrous tetrahydrofuran (250 ml) and stirred under nitrogen with cooling in an ice bath. Diethyl azadicarboxylate (5.6 ml, 35.6 mmol) was added dropwise. The reaction was allowed to warm to ambient temperature overnight. The volatiles were removed and the crude mixture dissolved in dichloromethane (50 ml) and filtered to remove excess phthalimide. The filtrate was concentrated in vacuo and the residue purified by silica gel chromatography in two portions using a gradient of ethyl acetate/hexane, (20% to 25% gradient) as eluent to give 7.1 g of 2-amino- 5-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-ylmethyI)-6-(4-methoxy-benzyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid terf-butyl ester. 1H-NMR (400 MHz, CDCI3) δ 7.87-7.84 (m, 2H), 7.75-7.73 (m, 2H), 7.07 (d, 2H, J = 8 Hz), 6.70 (d, 2H, J = 8 Hz), 5.94 (s, 2H), 4.09-4.05 (m, 1H), 3.84-3.73 (m, 2H), 3.76 (s, 3H), 3.66-3.43 (m, 4H), 2.89-2.83 (m, 1 H), 2.67-2.61 (m, 1 H), 1.53 (s, 9H) LC-MS: Rt = 3.03 min., m/z: 534.2 [M+H]+
A mixture of 2-amino-5-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-ylmethyl)-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (7.1 g, 13.3 mmol) in anhydrous ethanol (95 ml) was placed in a sonicator for complete dissolution. To this solution was added anhydrous hydrazine (1.92 ml, 58.7 mmol) and the reaction placed in an 80 °C bath with stirring for 6 hours, then allowed to cool to ambient temperature and stirred for an additional 2 days. The precipitate was filtered off and washed with ethanol (2 x 25 ml). The filtrate was evaporated to dryness under reduced pressure, the residue dissolved in dichloromethane (50 ml), filtered, and the solvent removed in vacuo. All volatile materials were removed under vacuum (0.5 x 10"6 mm Hg) at 50 °C for 6 hours, which afforded 5.26 g (98 %) of 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as a solid.
1H-NMR (400 MHz, CDCI3) δ 7.24 (d, 2H, J = 8 Hz), 6.86 (d, 2H, J = 8 Hz), 5.91 (s, 2H), 3.80 (s, 3H), 3.71-3.46 (m, 4H), 3.11-3.05 (m, 1 H), 2.98-2.93 (m, 1H), 2.87-2.72 (m, 2H), 2.53-2.47 (m, 1 H), 1.54 (s, 9H) LC-MS: Rt = 2.03 min, m/z: 404.2 [M+H]+
2-Amino-5-aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester (0.15 g, 0.37 mmol) was dissolved in a mixture of anhydrous dichloromethane (5 ml) and triethylamine (78 μl, 0.56 mmol). The flask was cooled in an ice-bath and stirred under nitrogen. Benzoyl chloride (43 μl, 0.37 mmol) was added dropwise and the reaction stirred for 5 minutes, then warmed to ambient temperature and stirred for an additional 16 hours. The solvent was removed in vacuo and the residue re-dissolved in ethyl acetate (20 ml). The organic phase was washed with 0.5 N hydrochloric acid (5 ml), saturated sodium bicarbonate (5 ml), and brine (5 ml). The solution was dried (MgSO4), filtered, and concentrated in vacuo to give 0.18 g of 2-amino-5- (benzoylamino-methyl)-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester.
1H-NMR (300 MHz, CDCI3) δ 7.77 (d, 2H, J - 7 Hz), 7.51-7.42 (m, 3H), 7.21 (d, 2H, J = 8 Hz), 7.00-6.98 (m, 1H), 6.84 (d, 2H, J = 8 Hz), 5.96 (s, 2H), 3.78 (s, 3H), 3.75-3.50 (m, 5H), 3.40-3.23 (m, 2H), 2.92 (dd, 1 H, J = 18 Hz and J = 6 Hz), 2.60 (dd, 1 H, J = 18 Hz and J = 6 Hz), 1.55 (s, 9H) LC-MS: Rt = 1.29 min, m/z: 509 [M+H]+
To a solution of crude 2-amino-5-(benzoylamino-methyl)-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (0.18 g, 0.35 mmol) in anhydrous dichloromethane (5 ml) was added imidazol-1-yl-oxo- acetic acid ferf-butyl ester (0.20 g, 1.05 mmol). The reaction was. stirred at ambient temperature for 16 hours. The solution was concentrated in vacuo and the residue purified by silica gel chromatography using a gradient of ethyl acetate/dichloromethane (5% to 10% gradient) as eluent, which afforded 0.18 g of 5-(benzoylamino-methyl)-2-(terf-butoxyoxalyl-amino)-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester. The overall yield for the first two steps was 77 %.
1H-NMR (300 MHz, CDCI3) δ 12.54 (s, 1 H), 7.76 (d, 2H, J = 7 Hz), 7.51-7.42 (m, 3H), 7.19 (d, 2H, J = 8 Hz), 6.99-6.96 (m, 1 H), 6.83 (d, 2H, J = 8 Hz), 3.87-3.68 (m, 4H), 3.77 (s, 3H), 3.58-3.53 (m, 1H), 3.40-3.29 (m, 2H), 2.99 (bd, 1H, J = 18 Hz), 2.68 (bd, 1H, J = 18 Hz), 1.60 (s, 9H) LC-MS: R, = 3.57 min, m/z: 636 [M+H]+
5-(Benzoylamino-methyl)-2-(førf-butoxyoxalyl-amino)-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (0.18 g, 0.283 mmol) was dissolved in a solution of 10 % formic acid/methanol (4 ml). Palladium on activated carbon (10 %, 90 mg) was added and the reaction stirred at ambient temperature for 24 hours. Another 70 mg of catalyst was added and the reaction stirred for an additional 72 hours. The solution was filtered through celite and the filter cake washed with hot methanol. The filtrate was concentrated In vacuo and the residue precipitated from hexanes. The solid material was filtered off and washed further with hexane affording 90 mg (62 %) of 5-(benzoyI- amino-methyl)-2-(terf-butoxyoxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as a solid.
1H-NMR (400 MHz, CDCI3) δ 12.49 (s, 1 H), 8.23 (bs, 1H), 7.97 (bs, 1H), 7.85 (d,
2H, J = 7 Hz), 7.49-7.38 (m, 3H), 4.17-4.04 (m, 2H), 3.99-3.94 (m, 1H), 3.65-3.59
(m, 1 H), 3.47-3.41 (m, 1 H), 3.22 (d, 1 H, J = 16 Hz), 2.81 (dd, 1 H, J = 16 Hz and
J = 10 Hz), 1.61 (s, 9H), 1.59 (s, 9H)
LC-MS: Rt = 1.26 min, m/z: 404 (loss of two t-butyl groups) [M+H]+
5-(Benzoyl-amino-methyl)-2-(ferf-butoxyoxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (60 mg, 0.12 mmol) was dissolved in a solution of 50 % trifluoroacetic acid/dichloromethane (3 ml) and stirred at ambient temperature for 16 hours. Adding the trifluoroacetic acid solution dropwise to diethyl ether precipitated the product. The suspension was stirred for 1 hour, filtered and washed with diethyl ether, which afforded 36 mg (60 %) of the title compound as a solid.
1H-NMR (400 MHz, DMSO) δ 12.34 (bs, 1H), 9.24 (bs, 2H), 8.86 (s, 1 H), 7.89 (d, 2H, J = 7 Hz), 7.56-7.46 (m, 3H), 4.39-4.24 (m, 2H), 3.68-3.59 (m, 3H), 3.24-3.18 (m, 2H, partially obscured by water), 2.89-2.83 (m, 1H), 2.65-2.32 (m, 1H, partially obscured by DMSO) LC-MS: Rt = 0.63 min, m/z: 404 [M+H]+
EXAMPLE 2
Figure imgf000031_0001
5-(((6-Bromo-2-p-tolyl-quinoline-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)- 2-(oxalyl-amino)-4,5.6,7-tetrahvdro-thienof2.3-clpyridine-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using 6-bromo-2-p-tolyl-quinoline-4-carboxylic acid and 2-amino-5-aminomethyl- 6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 745.2 [M+H]+ Calculated for C36H3ιN O7SBr, C2HF3O2; C, 53.22%; H, 3.76%; N, 6.65%. Found: C, 52.96%; H, 4.04%; N, 6.63%.
EXAMPLE 3
Figure imgf000032_0001
5-(((5-Benzyloxy-1H-indole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2- (oxalyl-amino)-4.5,6.7-tetrahvdro-thienor2.3-c1pyridine-3-carboxylic acid , The title compound was prepared in a similar way as described in Example 1 using 5-benzyloxy-indole-2-carboxylic acid and 2-amino-5-aminomethyl-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 669.4 [M+H]+
Calculated for C35H32N4O8S, 0.5xC2HF3O2, H2O; C, 58.14%; H, 4.68%; N, 7.53%. Found: C, 58.13%; H, 4.61%; N, 7.50%. EXAMPLE 4
Figure imgf000033_0001
5-((4-Benzoylamino-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4.5,6.7-tetrahvdro-thienor2.3-c1pyridine-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using 4-(benzoylamino)benzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 643.2 [M+H]+
Calculated for C33H3oN4O8S, 2xC2HF3O2; C, 51.04%; H, 3.70%; N, 6.43%. Found: C, 50.74%; H, 3.98%; N, 6.49%.
EXAMPLE 5
Figure imgf000033_0002
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4.5.6,7-tetrahydro- thienoF2,3-clpyridine-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using 4-(benzoylamino)-benzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
LC-MS: m/z: 523.0 [M+H]+
Calculated for C25H22N4O7S, 2/3xC2HF3O2, H2O; C, 51.80%; H, 3.96%; N, 9.18%. Found: C, 51.99%; H, 4.32%; N, 8.94%. EXAMPLE 6
Figure imgf000034_0001
6-(4-Methoxy-benzyl)-5-(((5-methyl-2-phenyl-2H-n .2.31triazole-4-carbonvn- amino)methyl)-2-(oxalyl-amino)-4,5.6.7-tetrahvdro-thienor2.3-clpyridine-3- carboxylic acid
The title compound was prepared in a similar way as described in Example 1 using 5-methyl-2-phenyl-2A-/-[1 ,2,3]triazole-4-carboxylic acid and 2-amino-5- aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 605.2 [M+H]+ Calculated for C29H28N6O7S, 1.5xC2HF3O2, H2O; C, 48.43%; H, 4.00%; N, 10.59%. Found: C, 48.72%; H, 4.19%; N, 10.18%.
EXAMPLE 7
Figure imgf000034_0002
5-((4-Ethoxy-2-hvdroxy-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4.5.6.7-tetrahvdro-thienor2.3-c1pyridine-3-carboxylic acid
The title compound was prepared in a similar way as described in Example 1 using 2-hydroxy-4-ethoxybenzoic acid and 2-amino-5-aminomethyl-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester as the starting material. The hydrogenation step in Example 1 was omitted. LC-MS: m/z: 585.2 [M+H]+ Calculated for C36H3ιN4O7SBr, C2HF3O2, 2/3xH2O; C, 48.19%; H, 4.24%; N, 5.44%. Found: C, 47.99%; H, 4.26%; N, 5.27%.
EXAMPLE 8
Figure imgf000035_0001
5-(((Biphenyl-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4,5.6.7-tetrahvdro-thienor2.3-clpyridine-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using 4-phenylbenzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 601.1 [M+H]+ Calculated for C32H29N3O S, 2xC2HF3O2, 2xH2O; C, 50.06%; H, 4.08%; N, 4.86%. Found: C, 50.20%; H, 3.83%; N, 4.70%.
EXAMPLE 9
Figure imgf000035_0002
5-(((Biphenyl-4-carbonyl)amino)methyl)-2-(oxalyl-amino)-4.5.6.7-tetrahvdro- thienor2,3-clpyridine-3-carboxylic acid
The title compound was prepared in a similar way as described in Example 1 using 4-phenylbenzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. LC-MS: m/z: 480.2 [M+H]+ HPLC (A1): Rt = 25.81 min.
EXAMPLE 10
Figure imgf000036_0001
5-((4-Ethoxy-2-hvdroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4.5.6.7- tetrahvdro-thienor2.3-clpyridine-3-carboxylic acid
The title compound was prepared in a similar way as described in Example 1 using 2-hydroxy-4-ethoxybenzoic acid and 2-amino-5-aminomethyl-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester as the starting material.
LC-MS: m/z: 464.1 [M+H]+ HPLC (A1 ): Rt = 19.98 min.
EXAMPLE 11
Figure imgf000036_0002
5-f(f5-Methyl-2-phenyl-2H-ri .2.31triazole-4-carbonyl)aminolmethyl)-2-(oxalyl- amino)-4,5,6.7-tetrahvdro-thienor2.3-clpyridin-3-carboxylic acid
The titie compound was prepared in a similar way as described in Example 1 using 5-methyI-2-phenyl-2H-[1 ,2,3]triazole-4-carboxylic acid and 2-amino- 5- aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material. LC-MS: m/z: 485.2 [M+H]+ HPLC (A1): Rt = 22.04 min
EXAMPLE 12
Figure imgf000037_0001
5-(f3-Biphenyl-4-yl-acryloylamino)methvπ-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4.5.6.7-tetrahvdro-thienof2.3-clpyridin-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using 3-(biphenyl-4-yl)acrylic acid and 2-amino-5-aminomethyl-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted. LC-MS: m/z: 626.2 [M+H]+ HPLC (A1): Rt = 35.60 min.
EXAMPLE 13
Figure imgf000037_0002
5-(((1H-lndole-2-carbonyl)-amino1-methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4.5.6.7-tetrahvdro-thienor2.3-clpyridin-3-carboxylic acid The title compound was prepared in a similar way as described in Example 1 using indole-2-carboxylic acid and 2-amino-5-aminomethyl-6-(4-ethoxybenzyl)- 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 563.2 [M+H]+ HPLC (A1): Rt = 27.59 min.
EXAMPLE 14
Figure imgf000038_0001
5-(((1H-lndole-2-carbonvπ-aminol-methyl>-2-(oxalyl-amino)-4.5.6 -tetrahvdro- thienor2,3-clpyridine-3-carboxylic acid
The titie compound was prepared in a similar way as described in Example 1 using 1H-indole-2-carboxylic acid and 2-amino-5-aminomethyl-6-(4-ethoxy- benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
LC-MS: m/z: 443.0 [M+H]+ HPLC (B1): Rt = 19.67 min. Calculated for C20Hι8N4O6S, 4xC2HF3O2 C, 37.43%; H, 2.47%; N, 6.24%. Found: C, 37.32%; H, 2.93%; N, 6.12%
EXAMPLE 15
Figure imgf000038_0002
5-((3-Biphenyl-4-yl-propionylamino)methyl)-2-(oxalyl-amino)-4.5.6.7-tetrahvdro- thieno[2.3-clpyridine-3-carboxylic acid
The titie compound was prepared in a similar way as described in Example 1 using 3-(biphenyl-4-yl)acrylic acid and 2-amino-5-aminomethyl-6-(4-methoxy- benzyI)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. LC-MS: m/z: 508,4 [M+Hf HPLC (A1): Rt = 22,23 min;
EXAMPLE 16
Figure imgf000039_0001
6-(4-Methoxy-benzyl)-5-(((naphthalene-1-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5.6.7-tetrahvdro-thienor2.3-clpyridine-3-carboxylic acid
The titie compound was prepared in a similar way as described in Example 1 using 1-naphthyI-carboxylic acid and 2-amino-5-aminomethyl-6-(4-ethoxy- benzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. The hydrogenation step in Example 1 was omitted.
LC-MS: m/z: 574.2 [M+H]+ HPLC (A1): Rt = 27.36 min
EXAMPLE 17
Figure imgf000040_0001
5-(Benzyloxycarbonylamino-methyl)-2-(oxalyl-amino)-4.5.6.7-tetrahvdro- thienof2.3-c1pyridine-3-carboxylic acid and 7-(benzyloxycarbonylamino-methyl)-2- (oxalyl-amino)-4.5.6.7-tetrahvdro-thienor2,3-c1pyridine-3-carboxylic acid In a 50 ml round-bottom flask, 2-(isoindole-1 ,3-dione-2-ylmethyl)-4-(2- spiro[1 ,3]dioxolane)-piperidine-1 -carboxylic acid ferf-butyl ester (543 mg, 1.35 mmol) was dissolved in 20 % trifluoroacetic acid in dichloromethane (20 ml). After stirring in for 25 hours at room temperature, dichloromethane (30 ml) was added to dilute the solution. Solid sodium bicarbonate was then slowly added to neutralize the solution, which was then extracted with dichloromethane (3 x 40 ml). The combined organic extracts were dried (MgSO ), filtered, and the solvent evaporated in vacuo. which afforded 385 mg (94%) of 2-(isoindole-1 ,3-dione-2- ylmethyl)-4-(2-spiro[1 ,3]dioxolane)-piperidine as a foam. 1H-NMR (300 MHz, CDCI3) δ 7.88-7.82 (m, 2H), 7.76-7.70 (m, 2H), 5.02-4.88 (bs, 1 H), 4.66-4.55 (bs, 1 H), 4.40-4.24 (bs, 1 H), 4.08-3.08 (m, 8H), 2.26-1.75 (m, 2H).
In a 25 ml round-bottom flask, 2-(isoindole-1 ,3-dione-2-ylmethyl)-4-(2- spiro[1 ,3]dioxolane)-piperidine (385 mg, 1.27 mmol) was suspended in absolute ethanol (13 ml), flushed with nitrogen and sealed with a rubber septum. The reaction mixture was stirred at room temperature. Hydrazine (160 μl, 5.1 mmol) was added via syringe. The reaction mixture was heated to 80 °C for 6 hours and stirred at room temperature for 14 hours. The white precipitate was filtered off and washed with absolute ethanol. The filtrate was concentrated in vacuo affording an oil, which was dissolved in dichloromethane (10 ml) and re- concentrated in vacuo affording 214 mg (98%) of 2-aminomethyl-4-(2- spiro[1 ,3]dioxolane)-piperidine as a solid.
1H-NMR (300 MHz, CDCI3) δ 3.87 (s, 4H), 3.60-3.29 (bs, 2H), 3.27-3.01 (m, 2H), 2.98-2.79 (m, 2H), 2.78-2.67 (m, 1 H), 1.95-1.62 (m, 3H), 1.61-1.45 (m, 1 H). In a 25 ml round-bottom flask, 2-aminomethyl-4-(2-spiro[1 ,3]dioxolane)-piperidine (553 mg, 3.21 mmol) in dichloromethane (10 ml) was flushed with nitrogen and sealed with a rubber septum. This solution was cooled to 0 °C before 95% benzyl chloroformate (550 μl, 3.9 mmol) was added dropwise. The solution was concentrated in vacuo and the residue was redissolved in 2N aqueous hydrochloric acid (12 ml) and heated to 50 °C with stirring for 15 hours. The reaction mixture was cooled to room temperature before dichloromethane (13 ml) and BOC-anhydride (11 ml, 10.4 g, 48 mmol) were added. Solid sodium bicarbonate was added slowly until the pH of the solution was 8. The bi-phasic mixture was stirred briskly at room temperaturet for 5 hours, extracted with dichloromethane (2 x 30 ml). The combined organic extracts were washed with brine (25 ml), dried (MgSO4), filtered, and the solvent evaporated in vacuo. The residue was purified by silica gel chromatography using a mixture of hexanes/ethyl acetate (3:1), which afforded 430 mg (37 %) of 2- (benzyloxycarbonyl-amino-methyl)-4-oxo-piperidine-1 -carboxylic acid ferf-butyl ester as an oil.
1H-NMR (300 MHz, CDCI3) δ 7.40-7.30 (bs, 5H), 5.09 (s, 2H), 4.74-4.63 (bs, 1 H), 4.36-4.16 (bs, 1H), 3.50-3.10 (m, 3H), 2.66 (dd, 1H, J = 16 Hz and J = 7 Hz), 2.56-2.29 (m, 3H), 1.46 (s, 9H).
In a 50 ml round-bottom flask, 2-(benzyloxycarbonyl-amino-methyl)-4-oxo- piperidine-1 -carboxylic acid ferf-butyl ester (430 mg, 1.19 mmol) in absolute ethanol (15 ml) was flushed with nitrogen and sealed with a rubber septum. Sulfur (44 mg, 1.4 mmol) and ferf-butyl cyano acetate (218 μl, 1.5 mmol) were added. The solution was cooled to 0 °C before morpholine (210 μl, 2.4 mmol) was added. Stirring was commenced as the reaction was heated to 50 °C for 20 hours. The cooled reaction mixture was concentration in vacuo and the residual oil was purified by silica gel chromatography using a mixture of hexanes/ethyl acetate (6:1) as eluent affording 474 mg (77 %) of a 1:1 mixture of 2-amino-7- (benzyloxycarbonylamino-methyl)-4,7-dihydro-5W-thieno[2,3-c]pyridine-3,6- dicarboxylic acid di-ferf-butyl ester and 2-amino-5-(benzyloxy-carbonylamino- methyl)-4,7-dihydro-5AV-thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester as a solids. 1H-NMR (300 MHz, CDCI3) δ 7.40-7.32 (m, 5H), 5.42-2.41 (m, 9H), 1.55 (s, 9H), 1.46 (s, 4.5H), 1.45 (s, 4.5H). In a 10 ml scintillating vial a solution of 2-amino-7-(benzyloxycarbonylamino- methyl)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester and 2-amino-5-(benzyloxy-carbonylamino-methyl)-4,7-dihydro-5H- thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester (193 mg, 0.37 mmol) in dichloromethane (4 ml) was treated with imidazol-1-yl-oxo-acetic acid ferf-butyl ester (520 mg, 2.7 mmol) in dichloromethane (2 ml). After stirring for 3 hours at room temperature, the reaction solution was concentrated in vacuo. The residue was purified by silica gel chromatography using a mixture of hexanes/ethyl acetate (6:1), which afforded 192 mg (77 %) of a mixture of 5-(benzyloxy- carbonylamino-methyl)-2-(ferf-butoxyoxalyl-amino)-4,7-dihydro-5H-thieno[2,3- c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester and 7-(benzyloxy- carbonylamino-methyl)-2-(terf-butoxyoxalyl-amino)-4,7-dihydro-5H-thieno[2,3- c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester as a foam.
LC-MS (APCI+) m/z: 646.3 [M+H]+;
In a 10 ml round-bottom flask, a mixture of 5-(benzyloxy-carbonylamino-methyl)- 2-(ferf-butoxyoxalyI-amino)-4,7-dihydro-5H-thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf-butyl ester and 7-(benzyloxy-carbonylamino-methyl)-2-(ferf-butoxy- oxalyl-amino)-4,7-dihydro-5 - -thieno[2,3-c]pyridine-3,6-dicarboxylic acid di-ferf- butyl ester (46 mg, 0.07 mmol) was dissolved in 30 % trifluoroacetic acid in dichloromethane (4 ml). The solution was left open to the atmosphere without stirring. After 24 hours a precipitate was filtered off and washed with diethyl ether, affording 35 mg (90 %) of the title compounds as a solid. 1H-NMR (300 MHz, DMSO) δ 12.35 (bs, 1H), 9.28 (bs, 2H), 7.79 (s, 0.5H), 7.68 (s, 0.5 H), 7.42-7.30 (m, 5 H), 5.09 (s, 1H), 5.06 (s, 1H), 4.57 (bs, 0.5H), 4.30 (q, 1H, J = 6 Hz), 3.60-2.70 (m partially obscured by water, 7H).
EXAMPLE 18
Figure imgf000043_0001
5-((2-Hvdroxy-benzoylamino)-methyl)-2-(oxalyl-amino)-4,5.6.7-tetrahvdro- thienoF2.3-clpyridine-3-carboxylic acid
Salicylic acid (1.25 g, 9.05 mmol) was dissolved in anhydrous N,N-dimethyl- formamide (15 ml) and placed under nitrogen atmosphere. Sodium hydride (0.76 g, 18.1 mmol) was added and the reaction stirred for 10 minutes, then benzyl bromide (4.3 ml, 36.2 mmol) was added. The reaction was stirred at ambient temperature for 4 days. The reaction mixture was diluted with ethyl acetate (75 ml) and washed with water (3 x 25 ml) followed by brine (15 ml). The organic layer was dried (MgSO ), filtered, and the solvent evaporated in vacuo. The residue was purified by silica gel chromatography affording 2.61 g (90 %) of 2- benzyloxy-benzoic acid benzyl ester as an oil.
1H-NMR (400 MHz, CDCI3) δ 7.86 (dd, 1H, J = 8 Hz and J = 2 Hz), 7.46-7I39 (m, 5H), 7.36-7.30 (m, 6H), 7.03-6.97 (m, 2H), 5.35 (s, 2H), 5.16 (s, 2H). LC-MS: Rt = 4.03 min, m/z: 319 [M+H]+
2-BenzyIoxy-benzoic acid benzyl ester (2.61 g, 8.15 mmol ) was dissolved in a solution of methanol (15 ml) and tetrahydrofuran (7 ml). Sodium hydroxide (0.49 g, 12.22 mmol) was dissolved in water (2 ml) and added to the reaction mixture. The reaction was stirred at ambient temperature for 24 hours, quenched with 6N hydrochloric acid to a final pH of 5. The mixture was evaporated to dryness in vacuo and the residue extracted into ethyl acetate (50 ml). The organic layer was extracted with 1 N sodium hydroxide (30 ml) and the layers separated. The aqueous layer was washed with dichloromethane (2 x 10 ml), acidified with 6N hydrochloric acid and extracted with ethyl acetate (2 x 30 ml). The combined organic extracts were washed with brine (15 ml), dried (MgSO ), filtered and the solvent evaporated in vacuo to give 1.71 g (92 %) of 2-benzyloxy-benzoic acid as an oil 1H-NMR (400 MHz, CDCI3) δ 8.20 (dd, 1H, J = 8 Hz and J = 2 Hz), 7.58-7.53 (m, 1H), 7.45-7.40 (m, 5H), 7.17-7.12 (m, 2H), 5.30 (s, 2H) LC-MS: Rt = 1.21 min, m/z: = 229 [M+H]+
To a solution of 2-benzyloxy-benzoic acid (0.12 g, 0.53 mmol) and 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.10 g, 0.62 mmol) in anhydrous acetonitrile (5 ml) was added triethylamine (0.18 ml, 1.6 mmol), followed by 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (0.176 g, 0.44 mmol). The reaction was stirred under nitrogen atmosphere for 16 hours. The solvents were removed in vacuo and the residue partitioned between ethyl acetate (30 ml) and water (10 ml). The layers were separated and the organic phase washed with 1% hydrochloric acid (10 ml), saturated sodium bicarbonate (10ml), and brine (10 ml). The organic layer was dried (MgSO4), filtered, and the solvent evaporated in vacuo. The residue was purified by silica gel chromatography using a mixture of methanol/dichloromethane (1 :19) as eluent, which afforded 82 mg (31 %) of 2- amino-5-((2-benzyloxy-benzoylamino)methyl)-6-(4-methoxy-benzyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as an oil. 1H-NMR (300 MHz, CDCI3) δ 8.43 (bs, 1 H), 8.23 (d, 1 H, J = 7 Hz), 7.46-7.40 (m, 1 H), 7.37-7.33 (m, 2H), 7.30-7.27 (m, 3H), 7.13-7.03 (m, 4H), 6.71 (d, 2H, J = 8 Hz), 5.95 (s, 2H), 5.20 (s, 2H), 3.82-3.74 (m, 1H), 3.76 (s, 3H), 3.61-3.29 (m, 4H), 3.12-3.06 (m, 2H), 2.85-2.77 (m, 1H), 2.57-2.50 (m, 1 H), 1.53 (s, 9H) LC-MS: R, = 1.39 min, m/z: 614 [M+H]+
To a solution of 2-amino-5-((2-benzyloxy-benzoylamino)methyl)-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (81 mg, 0.132 mmol) in anhydrous dichloromethane (4 ml) was added imidazol- 1-yl-oxo-acetic acid ferf-butyl ester (80 mg, 0.4 mmol). The reaction was stirred at ambient temperature for 3 hours and the solvent evaporated jn vacuo. The residue was purified by silica gel chromatography using a gradient of ethyl acetate/dichloromethane (8% to 11% gradient) as eluent, which afforded 97 mg (100 %) of 5-((2-benzyIoxy-benzoylamino)methyl)-2-(ferf-butoxyoxalyl-amino)-6- (4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester.
1H-NMR (400 MHz, CDCI3) δ 12.54 (s, 1 H), 8.41-8.39 (m, 1 H), 8.23 (dd, 1H, J = 8 Hz and J = 2 Hz), 7.46-7.41 (m, 1 H), 7.35-7.33 (m, 2H), 7.29-7.27 (m, 3H), 7.13-7.05 (m, 4H), 6.72 (d, 2H, J = 8 Hz), 5.19 (s, 2H), 3.84-3.75 (m, 1 H), 3.76 (s, 3H), 3.57-3.31 (m, 4H), 3.22-3.12 (m, 2H), 2.88 (dd, 1 H, J = 18 Hz and J = 5 Hz), 2.61 (dd, 1 H, J = 18 Hz and J = 5 Hz), 1.62 (s, 9H), 1.59 (s, 9H)
5-((2-Benzyloxy-benzoylamino)methyl)-2-(ferf-butoxyoxalyl-amino)-6-(4-methoxy- benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (97 mg, 0.13 mmol) was dissolved in a solution of 10 % formic acid/methanol (3 ml). Palladium on activated carbon (10 %, 25 mg) was added and the reaction stirred at ambient temperature for 24 hours. TLC analysis indicated that the reaction was incomplete. Additional catalyst (25 mg) was added and the reaction stirred for another 24 hours. Again, the reaction was judged incomplete by TLC and another 50 mg of catalyst was added and the reaction stirred for 72 hours. The solution was filtered through celite and the filter cake washed with hot methanol and ethyl acetate. The filtrate was concentrated jn vacuo, to the residue was added diethyl ether (4 ml) and the solution allowed to stand overnight. The precipitate was collected by filtration to give 12.5 mg (18 %) of 2- (ferf-butoxyoxalyl-amino)-5-((2-hydroxy-benzoylamino)methyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester.
1H-NMR (400 MHz, CDCI3) δ 12.47 (s, 1 H), 12.18 (s, 1H), 10.26-10.03 (m, 2H), 8.67 (bs, 1H), 7.94-7.91 (m, 1 H), 7.35-7.31 (m, 1H), 6.90 (d, 2H, J = 8 Hz), 4.46- 3.10 (m, 7H), 1.61 (s, 9H), 1.58 (s, 9H). LC-MS: Rt = 3.03 min, m/z: 532 [M+H]+
2-(ferf-Butoxyoxalyl-amino)-5-((2-hydroxy-benzoylamino)methyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (12 mg, 0.022 mmol) was dissolved in a solution of 50% trifluoroacetic acid/dichloromethane (1 ml) and stirred at ambient temperature for 16 hours. The trifluoroacetic acid solution was added dropwise to diethyl ether and stirred for 1 hour. The precipitate was filtered off, which afforded 10 mg (85 %) of the title compound as a solid. 1H-NMR (400 MHz, DMSO) δ 12.21 (s, 1H), 9.19-9.08 (m, 2H), 7.87-7.85 (m, 1 H), 7.42 (t, 1 H, J = 8 Hz), 6.92 (d, 2H, J = 8 Hz), 4.36-2.43 (m, 7H, partially obscured by water) LC-MS: Rt = 1.32 min, m/z: 420 [M+H]+
EXAMPLE 19
Figure imgf000045_0001
5-((3-Hvdroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4.5.6.7-tetrahvdro- thienor2,3-cl pyridine-3-carboxylic acid
A solution of 3-hydroxybenzoic acid (1.01 g, 7.3 mmol) was dissolved in anhydrous dichloromethane (20 ml) and placed under nitrogen atmosphere. , Diisopropylethylamine (5.7 ml, 32.9 mmol) was added, followed by additional dichloromethane (10 ml) for complete dissolution. The solution was cooled in an ice-bath and chloromethyl methyl ether (1.38 ml, 18.3 mmol) was added. The reaction was stirred and allowed to gradually warm to ambient temperature. The reaction mixture was then stirred at ambient temperature for 48 hours, the solvents evaporated in vacuo and the residue partitioned between ethyl acetate (75 ml) and water (25 ml). The layers were separated and the organic phase washed with 1% hydrochloric acid (3 x 15 ml), 1 N sodium hydroxide (2 x 10 ml), and brine (10 ml). The organic layer was dried (MgSO ), filtered, and evaporated in vacuo affording 1.2 g (73 %) of 3-methoxymethoxy-benzoic acid methoxymethyl ester as an oil.
1H-NMR (300 MHz, CDCI3) δ 7.73-7.70 (m, 2H), 7.36 (t, 1 H, J = 8 Hz), 7.26r7.22 (m, 1 H), 5.46 (s, 2H), 5.21 (s, 2H), 3.53 (s, 2H), 3.46 (s, 2H)
3-Methoxymethoxy-benzoic acid methoxymethyl ester (0.647 g, 2.86 mmol) was dissolved in methanol (10 ml). Sodium hydroxide (0.17 g, 7.15 mmol) dissolved in water (1 ml) was added to the solution. The reaction was stirred for 48 hours, concentrated to dryness in vacuo. The residue was dissolved in water (15 ml) and acidified with 1 N hydrochloric acid until a precipitate formed and the pH was 5. The aqueous solution was extracted with ethyl acetate (15 ml), acidified with additional 1 N hydrochloric acid, and extracted again with ethyl acetate (15 ml). The combined organic layers were washed with brine (10 ml), dried (MgSO4), filtered, and evaporated in vacuo to give 0.48 g (93 %) of 3-methoxy-methoxy- benzoic acid as a solid.
1H-NMR (400 MHz, CDCI3) δ 7.78-7.76 (m, 2H), 7.39 (t, 1 H, J = 8 Hz), 7.30-7.27 (m, 1 H), 5.24 (s, 2H), 3.50 (s, 2H)
To a solution of 3-methoxy-methoxy-benzoic acid (91 mg, 0.50 mmol) and 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (96 mg, 0.65 mmol) in anhydrous acetonitrile (4 ml) was added triethylamine (0.16 ml, 1.5 mmol), followed by 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (155 mg, 0.384 mmol). The reaction was stirred under nitrogen atmosphere for 16 hours. The volatiles were removed in vacuo and the residue partitioned between ethyl acetate (25 ml) and water (10 ml). The layers were separated and the organic phase washed with 0.5 N hydrochloric acid (2 x 5 ml), saturated sodium bicarbonate (5 ml), and brine (5 ml). The organic layer was dried (MgSO4), filtered, and concentrated in vacuo, which afforded 0.14 g of crude 2-amino-5-((3-methoxymethoxy- benzoylamino)methyl)-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as an oil.
1H-NMR (300 MHz, CDCI3) δ 7.47 (s, 1 H), 7.37-7.35 (m, 2H), 7.23-7.20 (m, 3H), 6.99-6.96 (bm, 1 H), 6.84 (d, 2H, J = 8 Hz), 5.96 (s, 2H), 5.22 (s, 2H), 3.78 (s, 3H), 3.75-3.54 (m, 5H), 3.48 (s, 3H), 3.42-3.23 (m, 2H), 2.95-2.87 (m, 1H), 2.63- 2.55 (m, 1 H) LC-MS: Rt = 3.07 min, m/z: 568 [M+H]+
To a solution of crude 2-amino-5-((3-methoxymethoxy-benzoylamino)-methyl)-6- (4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester (141 mg, 0.248 mmol) in anhydrous dichloromethane (5 ml) was added imidazol-1-yl-oxo-acetic acid ferf-butyl ester (0.15 g, 0.744 mmol). The reaction was stirred at ambient temperature for 16 hours, concentrated in vacuo and the crude material purified by silica gel chromatography using a gradient of ethyl acetate/dichloromethane (5% to 15% gradient) as eluent, which afforded 0.12 g of 2-(ferf-butoxyoxalyl-amino)-5-((3-methoxymethoxy- benzoylamino)methyl)-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- , c]pyridine-3-carboxylic acid ferf-butyl ester as an oil. The overall yield of the last two steps was 45%.
1H-NMR (300 MHz, CDCI3) δ 12.54 (s, 1 H), 7.47 (s, 1H), 7.37-7.35 (m, 2H), 7.21- 7.18 (m, 3H), 6.96-6.93 (bm, 1 H), 6.84 (d, 2H, J = 8 Hz), 5.22 (s, 2H), 3.87-3.67 (m, 4H), 3.78 (s, 3H), 3.58-3.54 (m, 1H), 3.48 (s, 3H), 3.42-3.32 (m, 2H), 3.82- 2.95 (m, 1 H), 2.71-2.64 (m, 1H), 1.60 (s, 18H) LC-MS: Rt = 1.47 min, m/z: 696 [M+H]+
2-(ferf-Butoxyoxalyl-amino)-5-((3-methoxymethoxy-benzoylamino)methyl)-6-(4- methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf- butyl ester (0.12 g, 0.17 mmol) was dissolved in a solution of 10% formic acid/methanol (4 ml). Palladium on activated carbon (10%, 0.10 g) was added and the reaction stirred at ambient temperature for 72 hours. The solution was filtered through celite and the filter cake washed with hot methanol and ethyl acetate. The filtrate was evaporated in vacuo and the residue was purified by radial silica gel chromatography (methanol/dichloromethane, 1%) affording 18 mg (18%) of 2-(ferf-butoxyoxalyl-amino)-5-((3-methoxymethoxy- benzoylamino)methyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester.
1H-NMR (400 MHz, CDCI3) δ 12.52 (s, 1 H), 7.49-7.48 (m, 1H), 7.42-7.33 (m, 2H), 7.19-7.16 (m, 1H), 6.86-6.84 (m, 1 H), 5.22 (s, 2H), 4.03-3.91 (m, 3H), 3.49 (s, 3H), 3.20-3.00 (m, 3H), 2.51-2.44 (m, 1H), 1.61 (s, 9H), 1.59 (s, 9H) LC-MS: Rt = 1.29 min, m/z: 576 [M+H]+
2-(ferf-ButoxyoxalyI-amino)-5-((3-methoxymethoxy-benzoylamino)methyl)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxyIic acid ferf-butyl ester (17 mg,
0.03 mmol) was dissolved in a solution of 50% trifluoroacetic acid/dichloromethane (1 ml) and stirred at ambient temperature for 16 hours.
Adding the trifluoroacetic acid solution dropwise to diethyl ether precipitated the product. The suspension was stirred for 1 hour, the solid filtered off and washed with diethyl ether, which afforded 8 mg (53 %) of the titie compound as a solid.
1H-NMR (400 MHz, DMSO-d6) δ 12.31 (s, 1H), 9.69 (s, 1H), 9.19 (bs, 2H), 8.73 (s, 1 H), 7.31-7.24 (m, 3H), 6.92 (d, 1 H, J = 8 Hz), 4.39-4.23 (m, 2H), 3.65-3.54
(m, 2H), 3.37-3.18 (m, 1 H, partially obscured by water), 2.87-2.81 (m, 1 H), 2.65-
2.43 (m, 1 H, partially obscured by DMSO)
LC-MS: Rt = 0.52 min, m/z: 420 [M+H]+
GENERAL CHIRAL SYNTHESIS
4-Oxo-l -((S)-1-phenyl-ethyl)-piperidine-(R)-2-carboxylic acid ethyl ester
Figure imgf000049_0001
Dichloromethane (11) and mol sieves 3 A (113 g) and amine (S)-(-)α-methyl- benzylamin (71 ,7 ml) were mixed in a 2 I three-necked bottle cooled to -5 °C (using a ethanol/water/ice bath). A 50% solution of ethylglyoxylate in toluene (117,6 ml) was added drop wise over 20 min., keeping the temperature between -5 °C and 0 °C The mixture was stirred for 0.5 hour before it was cooled to -30 °C. Trifluoroacetic acid (45,2 ml) was added over 3-4 minutes. Boron trifluoride diethyl ether (69,8 ml) was added drop wise over 5 minutes at -55 °C. The ice bath was removed and the mixture was allowed to warm up to -45 °C whereupon 2-(trimethylsilyloxy)-1 ,3-butadiene (100 ml) was added drop wise over 10 minutes. During the addition the mixture was cooled and the temperature kept below -20 °C. The above additions are all exothermic hence the cooling bath should have sufficient capacity to remove the heat generated during the rapid addition. The reaction mixture was stirred for 2 hours at -15 °C and 1 hour at 0 °C and then poured on ice/water and stirred for 15 minutes. Solid sodium hydrogen carbonate was added until pH 7-8. The mixture was stirred overnight at room temperature. The layers wee separated and the aqueous phase extracted with dichloromethane. The combined organic phases were filtered through a plug of silica eluting with dichloromethane. The relevant fractions were concentrated in vacuo. The residue was dissolved in hot heptane and cooled. This leaves a yellowish gummy material on the side of the flask and crystals starts forming. The heptane solution was heated again to dissolve crystals, leaving the gummy material on the side of the flask and the mixture was filtered hot. The heptane solution was cooled to room temperature and the precipitate was filtered off and dried in vacuo, which afforded 38 g of 4-oxo-1- ((S)-1-phenyl-ethyl)piperidine-(f?)-2-carboxylic acid ethyl ester as a solid. The filtrate was put in a refrigerator and a second crop was formed which was less pure and needed recrystallization from heptane to yield another 7,5 g of 4- oxo-1 -((S)-1-phenyl-ethyl)-piperidine-(R)-2-carboxylic acid ethyl ester.
4,4-Diethoxy-1 -((S)-1 -phenyl-ethyl)piperidine-(S)-2-carboxylic acid ethyl ester
Figure imgf000050_0001
The mother liquor from the above crystallization was concentrated m vacuo. 5.0 g of the resulting material (18.16 mmol) was dissolved in ethanol (100 ml) and triethylorthoformate (26.9 g, 181.6 mmol) and para-toluen-4-sulphonic acid (6.9 g, 36.32 mmol) was added. The reaction was stirred at room temperature for 16 hours before the mixture was poured on aqueous sodium hydrogen carbonate (200 ml) and extracted with ethyl acetate (4 x 75 ml). The combined extracts were concentrated in vacuo and purified by column chromatography (SiO2, Flash 40, petrol ether-ethyl acetate 10:1). Collection of the first band (Rf = 0.68) gave 1.14 g (18%) of 4,4-diethoxy-1-((S)-1-phenyl-ethyl)-piperidine-(f?)-2-carboxylic acid ethyl ester and collection of the second band (Rf = 0.4) gave 3.60 g (57%) of the title compound.
4,4-Diethoxy-1 -((S)-1 -phenyl-ethyl)-piperidine-( ?)-2-carboxylic acid ethyl ester
Figure imgf000050_0002
4-Oxo-1 -((S)-1-phenyl-ethyl)-piperidine-(R)-2-carboxylic acid ethyl ester (11.0 g, 0.040 mmol) was dissolved in a 1 :1 mixture of triethyl orthoformate and ethanol (140 ml) and para-toluene-4-sulphonic acid (15.2 g, 80 mmol) was added and the reaction mixture was stirred for 16 hours. The reaction mixture was neutralized with sodium bicarbonate (to pH 7-8), and extracted with dichloromethane (3 x 100 ml), dried (MgSO4), filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petrol ether/ethyl acetate 10:1), which afforded 12.0 g (86%) of the titie compound as an oil.
4,4-Diethoxy-1-((S)1-phenyl-ethyl)-(/?)-2-hydroxymethyl-piperidine
Figure imgf000051_0001
To a solution of 4,4-diethoxy-1-((S)-1-phenyl-ethyl)-piperidine-(/:?)-2-carboxylic acid ethyl ester (36.0 g, 0.103 mol) in dry diethyl ether (150 ml) was added a suspension of lithium aluminum hydride (5.88 g, 0.155 mol) in dry diethyl ether (300 ml) under an atmosphere of nitrogen at such a rate that the solution gently reflux. The reaction mixture was stirred over night before it was cooled to 0 °C and ethyl acetate (30 ml) was added drop wise to destroy excess lithium , aluminum hydride. After stirring for another 0.5 hour, water (12 ml) was added , drop wise. After stirring for 10-15 min the precipitate was filtered off through celite and the filter cage was washed with plenty of diethyl ether. The filtrate was washed with brine (100 ml), dried (MgSO4), filtered and the solvent evaporated m vacuo, which afforded 30 g (95%) of the titie compound as an oil.
4,4-Diethoxy-1-((S)-1-phenyl-ethyl)-( ?)-2-phthalimidomethyl-piperidine
Figure imgf000051_0002
A solution of 4,4-diethoxy-1-((S)1-phenyl-ethyl)-(f?)-2-hydroxymethyl-piperidine (65.35 g, 0.213 mmol), triphenylphosphine (61.3 g, 0.234 mol) and phthalimide (34.4 g, 0.234 mol) in tetrahydrofuran (700 ml) cooled to 0 °C was added diethyl azodicarboxylate over the course of 1.5 hour. The reaction mixture was stirred at 0 °C for another 2 hours before the solvent was removed in vacuo. The residue was dissolved in hot heptane-toluene (3:2) (650 ml) before it was cooled on an ice bath. The precipitate consisting of triphenyl phosphine oxide was filtered off and washed with heptane. The filtrate was concentrated in vacuo and the residue subjected to column chromatography using a mixture of toluene-ethyl acetate- heptane (3:1 :3) as eluent. The solvent was evaporated jn vacuo whereupon a , viscous oil was obtained. Upon addition of light petrol ether the product crystallized to give 67.4 g (73%) of the titie compound as a solid. 4,4-Diethoxy-1-((S)-1-phenyl-ethyl)-(S)-2-phthalimidomethyl-piperidine
Figure imgf000052_0001
A suspension of 4,4-diethoxy-1-((S)1-phenyl-ethyl)-(S)-2-hydroxymethyl- piperidine (20 g, 65 mmol), triphenylphosphine (18.76 g, 72 mmol) and phthalimide (10.52 g, 0.072 mol) in tetrahydrofuran (200 ml) cooled to 0 °C was added diethyl azodicarboxylate over the course of 1 hour. The reaction mixture was stirred at 0 °C for another 2 hours before the solvent was removed m vacuo. The residue was dissolve in a hot mixture of heptane/toluene (3:2) (100 ml) before it was cooled on an ice bath. The precipitate was filtered off and washed with heptane. The filtrate was concentrated vacuo and subjected to column chromatography using a mixture of toluene/ethyl acetate/heptane (3:1 :3) as eluent. The solvent was evaporated jn vacuo whereupon a viscous oil was obtained. Upon addition of light petrol ether (250 ml) the product crystallized to give 24 g (85%) of the titie compound as a solid.
4-Oxo-1-((S)-1-phenyl-ethyl)-2-(S)-phthalimidomethyl-piperidine
Figure imgf000052_0002
4,4-Diethoxy-1-(1-(S)-phenyl-ethyl)-2-(S)-phthalimidomethyl-piperidine (4.0 g, 9.2 mmol) was dissolved in a mixture of trifluoroacetic acid/water (9: 1 ) (100 ml) at 0 °C and stirred for 2 hours at this temperature. The mixture was basified with half saturated aqueous sodium carbonate, extracted with ethyl acetate and dried (MgSO4) for 2 hours and filtered. The solvent was removed in vacuo and the residue was left in a vacuum own at 40 °C for two days, which afforded 3.23 g (98%) of 4-oxo-l -((S)-1-phenyl-ethyl)-2-(S)-phthalimidomethyl-piperidine as a solid. 2-Amino-5-(S)-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-ylmethyl)-6-(1 -(S)-phenyl- ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester and 2-Amino-7-(R)-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-ylmethyl)-6-(1 -(S)-phenyl- ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester
Figure imgf000053_0001
A mixture of 4-oxo-1-(1-(f?)-phenyl-ethyl)-2-(S)-(phthalimidomethyl-piperidine (17.28 g, 47.73 mmol), ferf-butylcyanoacetat (7.41 g, 52.17 mmol), sulphur (1.71 g, 52.17 mmol) and morpholine (8.31 g, 95.46 mmol) in ethanol (150 ml) was heated under an atmosphere of nitrogen at 50 °C. The volatiles were removed in vacuo and the residue was subjected to column chromatography on silica gel using a mixture of heptane/ethyl acetate (5:1) as eluent. The band consisting of a mixture of 5- and 7-isomer was collected and purified on a reverse phase (C 8) column using a Flash 40 system. The residue was applied in a minimum volume of acetonitrile and eluted with a mixture of 40% acetonitrile in water containing 0.1 % trifluoroacetic acid. When the first isomer (the 5-isomer) was collected the eluent was changed to 50% acetonitrile in water with 0.1% trifluoroacetic acid and the 7-isomer was collected. Yield of: 2-amino-5-(S)-(1 ,3-dioxo-1 ,3-dihydro- isoindol-2-yImethyl)-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydrothieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester was 7.96 g. Yield of: 2-Amino-7-(/?)-(1 ,3-dioxo- ,3-dihydro-isoindol-2-ylmethyl)-6-(1-(S)- phenyl-ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester was 3.72 g (47% total of 5- and 7-isomer).
2-Amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester
Figure imgf000054_0001
2-Amino-5-(S)-(1 ,3-dioxo-1 ,3-dihydro-isoindol-2-ylmethyl)-6-(1-(S)-phenyl-ethyl)- 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (7.96 g, 15.4 mmol) and hydrazine hydrate (3.85 g, 77.0 mmol) in ethanol (250 ml) was stirred for 16 hours at room temperature. The solvent was removed jn vacuo and the white material obtained was extracted with diethyl ether (3 x 200 ml), the fractions were combined and the solvent was removed in vacuo to give 5.9 g (100%) of the titie compound as a solid.
Method A:
General procedure for amide formation:
To a solution of a relevant carboxylic acid (0.71 mmol) in Λ/,Λ/-dimethylformamide (5.0 ml) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (136 mg, 0.71 mmol) and [1 ,2,3]triazolo[4,5-b]pyridin-3-ol (96 mg, 0.71 mmol) and the mixture was stirred for 5 minutes at room temperature before a solution of diisopropylethylamine (121 DI, 0.71 mmol) and 2-amino-7-(S)-aminomethyl-6- (1 -(S)-phenyl-ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid tert- butyl ester (250 mg, 0.645 mmol) in Λ/,Λ/-dimethylformamide (5 ml) was added. The reaction was stirred 2 hours at room temperature before the solvent was removed in vacuo and the residue partitioned between ethyl acetate and water. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried (MgSO4), filtered and the solvent evaporated in vacuo. The residue was purified by column chromatography using a mixture of ethyl acetate and petroleum ether as eluent, which afforded pure amide derivatives.
Method B:
General procedure for formation of oxalamic acid ferf-butyl ester To a solution of the fused 2-amino-thiophene in dichloromethane was added imidazol-1-yl-oxo-acetic acid ferf-butyl ester (3 eq.) and triethylamine (3 eq.) and the reaction mixture was stirred 16 hours before it was transferred to a separation funnel and washed with aqueous sodium hydrogencarbonate. After drying (MgSO4) the solvent was removed in vacuo and the crude product was purified by column chromatography using a mixture of ethyl acetate and petroleum ether as eluent.
Method C: General procedure for hydrogenolysis of the N-α-methyl benzyl protection group
Method Ca:
The α-methyl benzyl protected amine was dissolved in a mixture of methanol/formic acid (9:1) (app. 10 ml/100 mg α-methyl benzyl protected amine). The solution was degassed by purging with nitrogen for 1 minute before 10% Pd/C (50% H2O content) was added. The reaction mixture was stirred at room temperature until TLC showed that all the starting material was consumed (typical 1-4 days). The Pd/C was filtered off using Celite and the filter cage was washed with plenty of methanol. The filtrate was concentrated in vacuo and the residue partitioned between ethyl acetate and water. The organic phase was separated, dried (MgSO4) and filtered. The solvent was removed in vacuo and the crude product was purified using column chromatography (SiO2, and a mixture of ethyl acetate/methanol as eluent).
Method Cb:
The α-methyl benzyl protected amine was dissolved in a mixture of isopropyl alcohol/formic acid (9:1) (app. 10 ml/100 mg α-methyl benzyl protected amine). The solution was degassed by purging with nitrogen for 1 minute before 10% Pd/C (dry) was added. The reaction mixture was stirred at room temperature until TLC showed that all the starting material was consumed (typical 1-4 days). The Pd/C was filtered off using Celite and the filter cage was washed with plenty of methanol. The filtrate was concentrated in vacuo and partitioned between ethyl acetate and water. The organic phase was separated, dried (MgSO4) and filtered. The solvent was removed in vacuo and the crude product was purified using column chromatography (SiO2, and a mixture of ethyl acetate/methanol as eluent)
Method D:
General procedure for deprotection of carboxylic acid di-ferf-butyl esters
The carboxylic acid ferf-butyl ester was dissolved in a mixture of trifluoroacetic acid/dichloromethane (1 :1) (app. 1 ml/100 mg carboxylic acid ferf-butyl ester). The reaction mixture was stirred for 16 hours at room temperature before diethyl ether (2 x the reaction volume) was added drop-wise. The precipitate was filtered off/spun down on a centrifuge and washed with diethyl ether to give analytical pure titie compound.
EXAMPLE 20
Figure imgf000056_0001
thienor2,3-clpyridine-3-carboxylic acid The titie compound was prepared using 3-phenoxy-benzoic acid and 2-amino-5- (S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3- carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
LC-MS: m/z: 496 [M+H]+, Rt = 2.00 min Calculated for C24H2ιN3O7S, 1.5xC2HF3O2 C, 48.65%; H, 3.40%; N, 6.30%. Found: C, 48.26%; H, 3.42%; N, 6.09%
EXAMPLE 21
Figure imgf000056_0002
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4.5.6.7-tetrahvdro- thienor2.3-clpyridine-3-carboxylic acid
The title compound was prepared using 4-benzoylamino-benzoic acid and 2- amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A,
B, C and D as described above.
Calculated for C25H22N4O7S, 1xH20, 1xC2HF3O2
C, 49.54%; H, 3.85%; N, 8.56%. Found: C, 49.26%; H, 3.96%; N, 8.27%
EXAMPLE 22
Figure imgf000057_0001
5-(f?)-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4.5,6,7- tetrahvdro-thienor2.3-clpyridine-3-carboxylic acid
The title compound was prepared using 4-benzoylamino-benzoic acid and 2- amino-5-(R)-aminomethyl-6-(1-(S)-phenyI-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
Calculated for C25H22N4O7S, 1xH2O, 1xC2HF3O2 C, 49.54%; H, 3.85%; N, 8.56%. Found: C, 49.79%; H, 4.13%; N, 8.42%
EXAMPLE 23
Figure imgf000057_0002
5-(S)-(((1-Hvdroxy-naphthalene-2-carbonvπamino)methvπ-2-(oxalyl-amino)- 4.5.6.7-tetrahvdro-thienor2,3-clpyridine-3-carboxylic acid The title compound was prepared using 1-hydroxy-naphthalene-2-carboxylic acid and 2-amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
Calculated for C22Hι9N3O7S, 1xH2O, 0.6xC2HF3O2 C, 50.13%; H, 3.92%; N, 7.56%. Found: C, 50.18%; H, 4.09%; N, 7.43%
EXAMPLE 24
Figure imgf000058_0001
5-(S)-(((3-Hvdroxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)- 4,5.6,7-tetrahvdro-thienof2,3-clpyridine-3-carboxylic acid The title compound was prepared using 3-hydroxy-naphthalene-2-carboxylic acid and 2-amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
Calculated for C22Hι9N3O7S, 1xH2O, 0.6xC2HF3O2 C, 50.13%; H, 3.92%; N, 7.56%. Found: C, 50.31%; H, 4.13%; N, 7.49%
EXAMPLE 25
Figure imgf000058_0002
5-(S)-(((3-Hvdroxy-7-methoxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5.6.7-tetrahvdro-thienof2,3-c1pyridine-3-carboxylic acid
The titie compound was prepared using 3-hydroxy-7-methoxy-naphthalene-2- carboxylic acid and 2-amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above. Calculated for C22Hι9N3O7S, 1xH2O, 0.7xC2HF3O2 C, 49.06%; H, 4.00%; N, 7.03%. Found: C, 49.30%; H, 4.37%; N, 6.66%
EXAMPLE 26 (52-1257)
Figure imgf000059_0001
2-(Oxalyl-amino)-5-(S)-((3-phenoxy-benzoylamino)methyl)-4,5,6,7-tetrahvdro- thienor2.3-clpyridine-3,6-dicarboxylic acid 6-acetoxymethyl ester
The title compound was synthesised using the above 3-phenoxy-berizoylamino compound and Method A, B, C as described above and the following steps
(S)-2-(ferf-Butoxyoxalyl-amino)-5-((3-phenoxy-benoylamino)methyl)-4,5,6,7- tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (100 mg, 0.165 mmol) and acetoxymethyl-p-nitrophenyl carbonate (46 mg, 0.182 mmol) was. heated to 50°C in acetonitrile (5 mL) for 6 hours. The temperature was lowered to 40°C and the reaction mixture was kept at this temperature for 3 days under N2. The solvent was removed in vacuo and the residue purified by flash column chromatography (SiO2, petrol ether-ethyl acetate (1 :1) affording 69 mg (58%) of (S)-2-(ferf-butoxyoxalyl-amino)-5-((3-phenoxy-benzoylamino)methyl)-4,7-dihydro- 5H-thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-acetoxymethyl ester 3-ferf-butyl ester as an oil.
(S)-2-(ferf-butoxyoxalyl-amino)-5-((3-phenoxy-benzoylamino)methyl)-4,7-dihydro- 5H-thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-acetoxymethyl ester 3-ferf-butyl ester (69 mg) was treated as described in Method D described above and the title compound was obtained in 69% yield. LC-MS: m/z: 612 [M+H]+, Rt = 3.80 min
EXAMPLE 27
Figure imgf000060_0001
2-(Oxalyl-amino)-5-(SM3-(4-phenoχy-phenyl)ureidomethyl)-4,5,6,7-tetrahydro- thienor2,3-clpyridin-3-carboxylic acid
To a solution of 2-amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester (500 mg, 1.29 mmol) in dichloromethane (5 ml) was added a solution of 4-phenoxyphenyl isocyanate (272 mg, 1.29 mmol) in dichloromethane (5 ml) over the course of 5 min. The reaction mixture was stirred for 45 min at room temperature before the solvent was removed in vacuo, which afforded 772 mg (100%) of 2-amino-5-(S)- (3-(4-phenoxy-phenyl)-ureidomethyl)-6-(1-(R)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester which was used without further purification.
The titie compound was obtained using the above ureido-compound and Method
B, C and D as described above.
LC-MS m/z: 511 [M+H]+, Rt = 2.13 min
Calculated for C24H22N4O7S, 1 x H2O and 1 x C2HF3O2
C, 48.60%; H, 3.92%; N, 8.72%. Found:
C, 48.95%; H, 3.80%; N, 8.50%
EXAMPLE 28
Figure imgf000060_0002
5-(R)-((4-Acetylamino-benzenesulfonylamino)methyl)-2-(oxalyl-amino)-4,5.6,7- tetrahvdro-thienor2.3-c1pyridine-3-carboxylic acid
The title compound was prepared using 4-acetylamino-benzenesulfonyI chloride and 2-amino-5-( ?)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. Formation of the sulpfonamide was performed in pyridine at room temperature using a slight excess of the sulfonyl chloride followed by Method B, C and D as described above.
Calculated for Cι9H20N4O8S2, 1 x C2HF3O2 C, 41.31%; H, 3.47%; N, 9.18%. Found: C, 42.59%; H, 4.21%; N, 9.04%
EXAMPLE 29
Figure imgf000061_0001
5-(S)-((4-Acetylamino-benzenesulfonylamino)methyl)-2-(oxalyl-amino)-4.5,6.7- tetrahvdro-thienof2.3-clpyridine-3-carboxylic acid
The titie compound was prepared using 4-acetylamino-benzenesulfonyl chloride and 2-amino-5-(S)-aminomethyl-6-(1 -(S)-phenyl-ethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
Formation of the sulfon amide was performed in pyridine at room temperature using a slight excess of the sulfonyl chloride followed by Method B, C and D as described above.
LC-MS: m/z: 497 [M+Hf, R, = 1.08 min Calculated for d9H20N4O8S2, 0.5 x H2O and 4/3 x C2HF3O2
C, 39.76%; H, 3.39%; N, 8.56%. Found:
C, 39.71%; H, 3.76%; N, 8.57%
EXAMPLE 30 (52-1267-1 A)
Figure imgf000061_0002
5-((4-Benzyl-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6.7-tetrahvdro- thienor2,3-clpyridine-3-carboxylic acid
The title compound was prepared in a similar way as described in Example 1 using 4-benzyl-benzoic acid and 2-amino-5-aminomethyl-6-(4-methoxy-benzyl)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material.
1H-NMR (300 MHz, DMSO-d6): δ 12.31 (bs, 1H), 9.2 (bs, 2H), 8.75 (bt, 1H), 7.72
(d, 2H), 7.35-7.14 (m, 6H), 4.44-4.24 (m, 2H), 3.99 (s, 2H), 3.73-3.57 (m, 3H),
3.22 (d, 1H), 2.86 (dd, 1 H).
LC-MS: m/z: 494 [M+H]+
EXAMPLE 31 (52-1268-1 A)
Figure imgf000062_0001
5-((5-Amino-5-methyl-hexanoylamino)methyl)-2-(oxalyl-amino)-4.5,6.7- tetrahvdro-thienor2.3-c1pyridine-3-carboxylic acid
The titie compound was prepared in a similar way as described in Example 1 using 5-ferf-butoxycarbonylamino-5-methyl-hex-2-enoic acid (prepared as described by Hansen et al. in J. Med. Chem. 41; (1998); 3705-3714) and 2- amino-5-aminomethyl-6-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material. 1H-NMR (300 MHz, DMSO-d6): δ 12.31 (bs, 1H), 9.3 (bs, 1H), 9.15 (bs, 1 H), 8.24 (bt, 1 H), 7.78 (bs, 3H), 4.42-4.24 (m, 2H), 3.59-3.38 (m, 3H), 3.19 (dd, 1H), 2.79 (dd, 1 H), 2.16 (bt, 2H), 1.52 (m, 4H), 1.2 (s, 6H). LC-MS: m/z: 427 [M+H]+
EXAMPLE 32 (1293-1 A)
Figure imgf000062_0002
5-(S)-(((Biphenyl-4-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6.7- tetrahvdro-thieno[2,3-c1pyridine-3-carboxylic acid
2-(4,4-Diethoxypiperidin-2-ylmethyl)isoindole-1 ,3-dione.
Figure imgf000063_0001
4,4-Diethoxy-1 -((S)-1 -phenyl-ethyl)-(S)-2-phthalimidomethyl-piperidine (15 g; prepared as in "General chiral synthesis") was dissolved in a mixture of ethanol (150 ml) and formic acid (15 ml). Palladium on charcoal (2 g; 10 %) was added under nitrogen and the mixture was stirred for 3 days. The solvent was removed in vacuo and the residue was treated with saturated aqueous sodium hydrogen- carbonate (100 ml) and methylene chloride (2 x 200 ml). The organic phase was dried (MgSO4), filtered and the solvent removed in vacuo affording 12.5 g of 2-, (4,4-diethoxypiperidin-2-ylmethyl)isoindole-1 ,3-dione. 1H-NMR (CDCI3, 400 Hz): δ 1.14 (m, 6H); 1.28 (m, 1H); 1.50 (dt, 1 H); 1.95 (m, 1H); 2.08 (m, 1H); 2.72 (dt, 1H); 2.90 (bs, 1 H); 3.05 (m, 1H); 3.17 (m, 1 H); 3.39 (t, 2H); 3.51 (t, 2H); 3.71 (m, 2H); 7.68 (m, 2H); 7.82 (m, 2H). LC-MS: m/z: 333.1 [M+H]+ 2-(4,4-Diethoxy-1-methylpiperidin-2-ylmethyl)isoindole-1,3-dione.
Figure imgf000063_0002
2-(4,4-diethoxypiperidin-2-ylmethyl)isoindole-1 ,3-dione (5.0 g 15 mmol) was dissolved in acetonitrile (50 ml) and treated with formaldehyde (1.13 ml, 15 mmol; 37 % in water) and sodium triacetoxyborohydride (4.45 g, 21 mmol) under nitrogen. The mixture was stirred for 2 hours and quenched with saturated aqueous sodium hydrogen carbonate (30 ml). The solvent was partly removed in vacuo and extracted with ethyl acetate (2 x 100 ml) and the organic phase was dried with magnesium sulphate and the solvent was removed in vacuo leaving 4.08 g of 2-(4,4-diethoxy-1-methylpiperidin-2-ylmethyl)isoindole-1 ,3-dione. 1H-NMR (CDCI3, 400 Hz): δ 1.15 (t, 6H); 1.45 (dd, 1H); 1.68 (dt, 1H); 1.95 (m, 2H); 2.40 (dt, 1H); 2.47 (s, 3H); 2.57 (m, 1H); 2.81 (m, 1H); 3.37 (m, 2H); 3.46 (q, 2H); 3.67; 3.95 (ab-syst.; 2H). LC-MS: m/z: 347.1 [M+H]+ The title compound was prepared from 2-(4,4-diethoxy-1-methylpiperidin-2- ylmethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and method A, B, C and D in "General chiral synthesis".
1H-NMR (300 MHz, DMSO-d6): δ 12.33 (s, 1H), 10.3 (bs, 1H), 8.8 (bs, 1H), 7.97
(d, 2H), 7.81 (d, 2H), 7.74 (d, 2H), 7.50 (t, 2H), 7.42 (t, 1H), 4.75-4.36 (m, 2H),
3.97-3.51 (m, 4H), 3.28 (dd, 1 H), 3.17-2.88 (m, 3H).
LC-MS: m/z: 494 [M+H]+
EXAMPLE 33 (52-1296-1 A)
Figure imgf000064_0001
5-(SH(4-Benzyl-benzoylamino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahvdro-thienor2,3-clpyridine-3-carboxylic acid
The titie compound was prepared from 2-(4,4-diethoxy-1-methylpiperidin-2- yImethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and method A, B, C and D in "General chiral synthesis"
Calculated for C26H25N3O6Sι, 0.5xH2O and 1xC2HF3O2 C, 53.3%; H, 4.3%; N, 6.6%. Found: C, 53.5%; H, 4.5%; N, 6.6%
1H-NMR (400 MHz, DMSO-d6): δ 12.31 (bs, 1 H), 8.72 (bt, 1 H), 7.80 (d, 2H), 7.35 (d, 2H), 7.30-7.18 (m, 5H), 4.49 (bs, 2H), 4.00 (s, 2H), 3.93-3.45 (m, 4H), 3.22 (d, 1 H), 3.09-2.89 (bm, 3H). LC-MS: m/z: 508 [M+H]+
EXAMPLE 34 (52-1297-1 A)
Figure imgf000064_0002
5-(S)-(((Naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4.5.6.7- tetrahvdro-thienor2.3-c1-pyridine-3-carboxylic acid The titie compound was prepared using naphthalene-2-carboxylic acid and 2- amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A, B, C and D as described above.
1H-NMR (400 MHz, DMSO-d6): δ 13.81 (bs, 1H), 12.32 (bs, 1H), 9.26 (bs, 2H), 9.06 (bt, 1H, -CONHCH2), 8.51 (s, 1H), 8.06-7.97 (m, 4H), 7.64-7.60 (m, 2H), 4.40 (dd, 2H), 3.79-3.66 (m, 3H), 3.52 (d, 1 H), 2.95 (dd, 1H). LC-MS: m/z: 454 [M+H]+
EXAMPLE 35 (52-1298-1 A)
Figure imgf000065_0001
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4.5.6,7-tetrahvdro- thienor2,3-clpyridine-3-carboxylic acid The title compound was prepared using 1/- -indole-2-carboxylic acid and 2- amino-5-(S)-aminomethyl-6-(1-(S)-phenyl-ethyl)-4,5,6,7-tetrahydro-thieno[2,3- c]pyridine-3-carboxylic acid ferf-butyl ester as the starting material and Method A,
B, C and D as described above.
1H-NMR (400 MHz, DMSO-d6): δ 13.78 (bs, 1 H), 12.32 (bs, 1H), 11.63 (s, 1 H), 9.29 (bs, 2H), 8.87 (bt, 1 H, -CONHCH2), 7.64 (d, 1 H), 7.45 (d, 1 H), 7.18 (m, 2H),
7.05 (m, 1H), 4.39 (dd, 2H), 3.71-3.63 (m, 3H), 3.26 (d, 1 H), 2.90 (dd, 1 H).
LC-MS: m/z: 443 [M+H]+
EXAMPLE 36 (52-1308-1 A)
Figure imgf000065_0002
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4.5.6.7- tetrahvdro-thienor2,3-c1pyridine-3-carboxylic acid
The titie compound was prepared from 2-(4,4-diethoxy-1-methylpiperidin-2- ylmethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and method A, B, C and D in "General chiral synthesis" 1H-NMR (400 MHz, DMSO-d6): δ 13.78 (bs, 1H), 12.34 (bs, 1 H), 11.67 (s, 1H), 10.33 (bs, 1H), 8.79 (bs, 1 H, -CONWCH2), 7.64 (d, 1 H), 7.44 (d, 1 H), 7.18 (m, 2H), 7.04 (m, 1 H), 4.46 (dd, 2H), 3.84-3.71 (m, 3H), 3.53 (bs, 1 H), 3.28 (d, 1H), 3.03 (m, 3H).
LC-MS: m/z: 457 [M+H]+
EXAMPLE 37 (52-1312-1 A)
Figure imgf000066_0001
6-Benzyl-5-(S)-(((naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4.5.6.7-tetrahvdro-thieno[2.3-clpyridine-3-carboxylic acid
The title compound was prepared from 2-(1-benzyl-4,4-diethoxy-piperidin-2- ylmethyl)isoindole-1 ,3-dione using the same methods as in Example 1 and 32 and "General chiral synthesis"
1H-NMR (400 MHz, DMSO-d6): δ 13.78 (bs, 1 H), 12.38 (bs, 1 H), 10.45 (bs, 1 H), 9.04 (bs, 1 H), 8.47 (bs, 1H, -CONWCH2), 8.00 (m, 4H), 7.63 (m, 2H), 7.48 (m, 3H), 4.31 (bs, 3H), 3.95 (bs, 3H), 3.66 (bs, 1H), 3.26 (bs, 2H). LC-MS: m/z: 544 [M+H]+
EXAMPLE 38 (52-1313-1 A)
Figure imgf000066_0002
6-Benzyl-5-(S)-(((1/-/-indole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4.5.6.7- tetrahvdro-thieno[2.3-clpyridine-3-carboxylic acid The titie compound was prepared from 2-(1-benzyl-4,4-diethoxy-piperidin-2- ylmethyl)isoindole-1 ,3-dione using the same methods as in example 1 and 32 and "General chiral synthesis"
1H-NMR (400 MHz, DMSO-d6): δ 13.8 (bs, 1H), 12.38 (bs, 1H), 11.69 (s, 1H), 10.4 (bs, 1 H), 8.86 (bs, 1H, -CONHCH2), 7.65 (d, 1H), 7.48 (m, 6H), 7.20 (t, 2H), 7.16 (s, 1 H), 7.05 (t, 1 H), 4.7 (bm, 2H), 4.32-4.20 (m, 3H), 3.85 (m, 1 H), 3.63 (m, 1H) 3.24 (bm, 2H). LC-MS: m/z: 533 [M+H]+

Claims

1. A compound of Formula 1
Figure imgf000068_0001
Formula 1
wherein X is -C(O)- or -S(O)2-;
Ri and R2 are independently hydrogen or a functional group that can be converted to hydrogen in vivo;
R3 is hydrogen, C -C6alkyl, H2N-R35-, C2-C6alkenyl, C2-C6alkynyl, aryl, aryl-Rι0-, aryl-N(R35)-, aryl-R! ι-N(R36)- , d-C6alkyloxy or aryI-Rι3-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, d-C6alkyl, R37-aryl-, R38-, R38-
Ri4-, Cι-C6alkyloxy, R38-O-, R38-Rι5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)-
N(R2i)- or R38-R23-C(O)-N(R24)-; R4 is hydrogen, Cι-C6alkyl, aryl-R26-, R27-O-C(O)-, aryl-R28-O-C(O)-, R29-C(O)-O-
R30-O-C(O)- or aryl-R3ι-C(O)-O-R32-O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, d-C6alkyloxy or R39-R34-O-; and wherein R5, R7, R8, R9, R10, Rn, Rι3, Rι4, R15, R23, R26, R28. R30. R31. 32> R33> R34 and R35 independently are d-C6alkylene, wherein R6, Rι2, R17, Rι8, R 0, R22,
R25, R27 and R29 independently are Cι-C6alkyl and wherein Rι6, Rι9, R21, R24, R35 and R36, and R37 independently are hydrogen or Cι-C6alkyl;
R38 and R39 are independently phenyl, naphthyl or thiophenyl
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 form.
2. A compound according to claim 1 wherein R^ and R2 are independently hydrogen, d-Cealkyl, aryl-R5-, R6-C(O)-O-R7- or aryl-R8-C(O)-O-R9- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, d-C6alkyl or d-C6alkyloxy;
3. A compound according to any one of the claims 1 to 2 wherein X is C(O).
4. A compound according to any one of the claims 1 to 2 wherein X is S(O)2.
5. A compound according to any one of the claims 1 to 4 wherein R and R2 are independently hydrogen, CrC6alkyl, aryl-R5-, or R6-C(O)-O-R7-, wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, trihalomethyl, d-C6alkyl or d-C6alkyloxy.
6. A compound according to claim 5 wherein Ri and R2 are independently hydrogen or CrC6alkyl.
7. A compound according to claim 6 wherein R1 and R2 are hydrogen.
8. A compound according to any one of the claims 1 to 7 wherein R3 is d- C6alkyl, C2-C6alkenyl, C2-C6alkynyl, aryl, aryl-Rι0-, CrC6alkyloxy, H2N-R35- or aryl-Rι3-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3- triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R38-Rι4-, CrC6alkyloxy, R38-O-, R38-Rι5- O-, R38-N(Rιe)-, Rιβ-C(O)-N(R1β)-f R38-C(O)-N(R2ι)- or R38-R23-C(O)-N(R24)-.
9. A compound according to claim 8 wherein R3 is d-Cealkyl, C2-C6alkenyl, aryl, H2N-R35- or aryl-Ri3-O- wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, d-Cealkyl, R37-aryl-, R38-, R38-Rι -, CrC6alkyloxy, R∞-O-, R38-Ri5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)-N(R21)- or R^-R^- C(O)-N(R24)-.
10. A compound according to claim 9 wherein R3 is CrC6alkyl or aryl, wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, d- Cealkyl, R37-aryl-, Ra,-, R38-Rι4-, d-C6alkyloxy, R38-O-, R38-Rι5-O-, R38-N(Rι6)-, Rι8-C(O)-N(Rι9)-, R38-C(O)-N(R2ι)- or R38-R23-C(O)-N(R24)-.
11. A compound according to claim 10 wherein R3 is aryl, wherein aryl is phenyl, biphenyl, indenyl, naphthyl, imidazolyl, 1 ,2,3-triazolyl, thiophenyl, pyridyl, quinolyl, isoquinolyl, indolyl or benzimidazolyl, which aryl group is optionally substituted with halogen, nitro, cyano, hydroxy, trihalomethyl, CrC6alkyl, R37-aryl-, R38-, R38- Ri4-, d-C6alkyloxy, R38-O-, R38-Rι5-O-, R38-N(Rι6)-, R18-C(O)-N(Rι9)-, R38-C(O)- N(R2ι)- or R38-R23-C(O)-N(R24)-.
12. A compound according to any one of the claims 8 to 11 wherein aryl is phenyl, biphenyl, naphthyl, 1 ,2,3-triazolyl, quinolyl or indolyl.
13. A compound according to claim 12 wherein aryl is phenyl, naphthyl, or indolyl.
14. A compound according to claim 13 wherein aryl is phenyl.
15. A compound according to any one of the claims 8 to 14 wherein the aryl is substituted by halogen, hydroxy, CrC6alkyl, R37-aryl-, R38-, R38-Rι4-, d-C6alkyl- oxy, R38-O-, R38-Ri5-O-, Ri8-C(O)-N(Ri9)-, or R38-C(O)-N(R2ι)-.
16. A compound according to claim 15 wherein the aryl is substituted by hydroxy, CrCealkyl, R38-, R 8-Rι4-, CrC6alkyloxy, R38-O-, R18-C(O)-N(Rι9)-, or R38-C(O)- N(R2ι)-.
17. A compound according to any one of the claims 8 to 16 wherein R38 is phenyl or thiophenyl.
18. A compound according to any one of the claims 1 to 17 wherein R is hydrogen, CrC6alkyl, aryl-R26-, R29-C(O)-O-R30-O-C(O)- or aryl-R3i-C(O)-O-R32- O-C(O)- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, Ci- Cealkyloxy or R39-R34-O-.
19. A compound according to claim 18 wherein R4 is hydrogen, d-C6alkyl, or aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, Ci- Cealkyloxy or R39-R3 -O-.
20. A compound according to claim 19 wherein R is hydrogen or aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, CrC6alkyloxy or R39-R34-O-.
21. A compound according to claim 20 wherein R4 is hydrogen.
22. A compound according to claim 20 wherein R4 is aryl-R26- wherein aryl is phenyl, naphthyl or thiophenyl, which aryl group is optionally substituted with halogen, nitro, cyano, trihalomethyl, R39-, R39-R33-, CrC6alkyloxy or R3g-R3.rO-
23. A compound according to any one of the claims 18 to 22 wherein aryl is phenyl.
24. A compound according to any one of the claims 18 to 23 wherein aryl is substituted by R39-.
25. A compound according to any one of the claims18 to 24 wherein R39 is Cr C6alkyloxy.
26. A compound according to claim 25 wherein R39 is methoxy.
27. A compound according to any one of the preceding claims selected from the following:
5-(Benzoylamino-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c] pyridine-3-carboxylic acid; 5-(((6-Bromo-2-p-tolyl-quinoline-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-
2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(((5-Benzyloxy-1 -/-indole-2-carbonyI)amino)methyl)-6-(4-methoxy-benzyl)-2-
(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-((4-Benzoylamino-benzoylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
6-(4-Methoxy-benzyl)-5-(((5-methyI-2-phenyl-2tf-[1 ,2,3]triazole-4- carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-
3-carboxylic acid;
5-((4-Ethoxy-2-hydroxy-benzoylamino)methyl)-6-(4-methoxy-benzyI)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(((Biphenyl-4-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(((Biphenyl-4-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Ethoxy-2-hydroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- , tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(((5-Methyl-2-phenyl-2H-[1 ,2,3]triazole-4-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-((3-Biphenyl-4-yl-acryloylamino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-(((1H-lndole-2-carbonyl)amino)methyl)-6-(4-methoxy-benzyl)-2-(oxalyl-amino)- 4,5,6,7-tetrahydro-thieno[2,3-c]pyridin-3-carboxylic acid;
5-(((1H-lndole-2-carbonyl)amino)methyI)-2-(oxaIyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((3-Biphenyl-4-yl-propionylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid; 6-(4-Methoxy-benzyI)-5-(((naphthalene-1-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(Benzyloxycarbonylamino-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- , thieno[2,3-c]pyridine-3-carboxylic acid;
7-(Benzyloxycarbonylamino-methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxyIic acid; 5-((2-Hydroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((3-Hydroxy-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c] pyridine-3-carboxylic acid; 2-(Oxalyl-amino)-5-(S)-((3-phenoxy-benzoylamino)methyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-(R)-((4-Benzoylamino-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((1-Hydroxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((3-Hydroxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(S)-(((3-Hydroxy-7-methoxy-naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl- amino)-4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
2-(Oxalyl-amino)-5-(S)-((3-phenoxy-benzoylamino)methyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3,6-dicarboxylic acid 6-acetoxymethyl ester;
2-(Oxalyl-amino)-5-(S)-(3-(4-phenoxy-phenyl)ureidomethyl)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridin-3-carboxylic acid;
5-((4-Benzyl-benzoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-((5-Amino-5-methyl-hexanoylamino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 5-(S)-(((Biphenyl-4-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-((4-Benzyl-benzoylamino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((Naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]-pyridine-3-carboxyIic acid;
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7-tetrahydro- thieno[2,3-c]pyridine-3-carboxylic acid;
5-(S)-(((1H-lndole-2-carbonyl)amino)methyl)-6-methyl-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid; 6-Benzyl-5-(S)-(((naphthalene-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-
4,5,6,7-tetrahydro-thieno[2,3-c]pyridine-3-carboxyIic acid; 6-Benzyl-5-(S)-(((1H-indole-2-carbonyl)amino)methyl)-2-(oxalyl-amino)-4,5,6,7- tetrahydro-thieno[2,3-c]pyridine-3-carboxylic acid;
or a pharmaceutically acceptable salt thereof.
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US6818787B2 (en) 2001-06-11 2004-11-16 Xenoport, Inc. Prodrugs of GABA analogs, compositions and uses thereof
US6833140B2 (en) 2001-06-11 2004-12-21 Xenoport, Inc. Orally administered dosage forms of GABA analog prodrugs having reduced toxicity
US8168623B2 (en) 2001-06-11 2012-05-01 Xenoport, Inc. Prodrugs of GABA analogs, compositions and uses thereof
US6972341B2 (en) 2001-06-11 2005-12-06 Xeno Port, Inc. Prodrugs of GABA analogs, compositions and uses thereof
US9238616B2 (en) 2001-06-11 2016-01-19 Xenoport, Inc. Prodrugs of gaba analogs, compositions and uses thereof
US7186855B2 (en) 2001-06-11 2007-03-06 Xenoport, Inc. Prodrugs of GABA analogs, compositions and uses thereof
US7232924B2 (en) 2001-06-11 2007-06-19 Xenoport, Inc. Methods for synthesis of acyloxyalkyl derivatives of GABA analogs
US7790708B2 (en) 2001-06-11 2010-09-07 Xenoport, Inc. Prodrugs of GABA analogs, compositions and uses thereof
US7423169B2 (en) 2001-06-11 2008-09-09 Xenoport, Inc. Methods for synthesis of acyloxyalkyl derivatives of GABA analogs
EP1534264A4 (en) * 2002-03-01 2007-06-13 Sunesis Pharmaceuticals Inc COMPOUNDS MODULATING THE ACTIVITY OF PTP-1B AND TC-PTP
WO2003082841A1 (en) 2002-04-03 2003-10-09 Novartis Ag 5-substituted 1,1-dioxo-`1,2,5!thiazolidine-3-one derivatives as ptpase 1b inhibitors
EP2341049A1 (en) 2002-04-03 2011-07-06 Novartis AG 5-substituted 1,1-dioxo-[1,2,5]thiazolidine-3-one derivatives as PTPASE 1B inhibitors
WO2004099192A3 (en) * 2003-04-30 2005-01-13 Inst Of Pharmaceutical Discove Heterocycle substituted carboxylic acids as inhibitors of protein tyrosine phosphatase-1b
US7329680B2 (en) 2003-04-30 2008-02-12 The Institute For Pharmaceutical Discovery, Llc Heterocycle substituted carboxylic acids
JP2006525366A (en) * 2003-04-30 2006-11-09 ジ インスチチュート フォー ファーマシューティカル ディスカバリー、エルエルシー Heterocyclic carboxylic acid substituent
US7475572B2 (en) * 2003-05-27 2009-01-13 Whirlpool Corporation Washing machine with a device for the security during transport
US8795725B2 (en) 2004-11-04 2014-08-05 Xenoport, Inc. GABA analog prodrug sustained release oral dosage forms
US8906412B2 (en) 2004-11-04 2014-12-09 Xenoport, Inc. GABA analog prodrug sustained release oral dosage forms
WO2006055708A3 (en) * 2004-11-18 2006-08-03 Inst For Pharm Discovery Inc Heterocycle substituted carboxylic acids for the treatment of diabetes
US8048917B2 (en) 2005-04-06 2011-11-01 Xenoport, Inc. Prodrugs of GABA analogs, compositions and uses thereof
WO2010118241A3 (en) * 2009-04-08 2014-03-20 Indiana University Research & Technology Corporation Inhibitors of protein tyrosine phosphatases
US9217012B2 (en) 2009-04-08 2015-12-22 Indiana University Research And Technology Corporation Inhibitors of protein tyrosine phosphatases
US10072043B2 (en) 2009-04-08 2018-09-11 Indiana University Research And Technology Corporation Inhibitors of protein tyrosine phosphatases
JP2021520415A (en) * 2018-04-04 2021-08-19 エピオダイン，インク． Opioid receptor modulators and related products and methods
JP7402857B2 (en) 2018-04-04 2023-12-21 エピオダイン，インク． Opioid receptor modulators and related products and methods

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