[go: up one dir, main page]

WO1999009006A1 - SYNTHESE DE CLASTO-LACTACYSTINE β-LACTONE ET DE SES ANALOGUES - Google Patents

SYNTHESE DE CLASTO-LACTACYSTINE β-LACTONE ET DE SES ANALOGUES Download PDF

Info

Publication number
WO1999009006A1
WO1999009006A1 PCT/US1998/016858 US9816858W WO9909006A1 WO 1999009006 A1 WO1999009006 A1 WO 1999009006A1 US 9816858 W US9816858 W US 9816858W WO 9909006 A1 WO9909006 A1 WO 9909006A1
Authority
WO
WIPO (PCT)
Prior art keywords
aryl
formula
alkyl
alkaryl
optionally substituted
Prior art date
Application number
PCT/US1998/016858
Other languages
English (en)
Inventor
François SOUCY
Louis Plamondon
Mark Behnke
William Roush
Original Assignee
Soucy Francois
Louis Plamondon
Mark Behnke
William Roush
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Soucy Francois, Louis Plamondon, Mark Behnke, William Roush filed Critical Soucy Francois
Priority to AU89062/98A priority Critical patent/AU749857B2/en
Priority to KR1020007001558A priority patent/KR20010022950A/ko
Priority to NZ503169A priority patent/NZ503169A/xx
Priority to BR9811304-6A priority patent/BR9811304A/pt
Priority to CA002301054A priority patent/CA2301054A1/fr
Priority to IL13453898A priority patent/IL134538A/xx
Priority to JP2000509690A priority patent/JP2001515064A/ja
Priority to EP98940885A priority patent/EP1021407A4/fr
Publication of WO1999009006A1 publication Critical patent/WO1999009006A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/08—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D263/16—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4015—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil having oxo groups directly attached to the heterocyclic ring, e.g. piracetam, ethosuximide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • C07C231/14—Preparation of carboxylic acid amides by formation of carboxamide groups together with reactions not involving the carboxamide groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/70—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/72—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms
    • C07C235/80—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups and doubly-bound oxygen atoms bound to the same carbon skeleton with the carbon atoms of the carboxamide groups bound to acyclic carbon atoms having carbon atoms of carboxamide groups and keto groups bound to the same carbon atom, e.g. acetoacetamides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C237/06—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C247/00—Compounds containing azido groups
    • C07C247/02—Compounds containing azido groups with azido groups bound to acyclic carbon atoms of a carbon skeleton
    • C07C247/04—Compounds containing azido groups with azido groups bound to acyclic carbon atoms of a carbon skeleton being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/66—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety
    • C07C69/67—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of saturated acids
    • C07C69/675—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of saturated acids of saturated hydroxy-carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/18—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member
    • C07D207/22—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/24—Oxygen or sulfur atoms
    • C07D207/26—2-Pyrrolidones
    • C07D207/273—2-Pyrrolidones with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to other ring carbon atoms
    • C07D207/277—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D207/28—2-Pyrrolidone-5- carboxylic acids; Functional derivatives thereof, e.g. esters, nitriles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the invention relates generally to methods for preparing lactacystin and related compounds, to novel analogs of lactacystin and c/ ⁇ sto-lactacystin ⁇ - lactone, and their uses as proteasome inhibitors.
  • the Streptomyces metabolite lactacystin (1) inhibits cell cycle progression and induces neurite outgrowth in cultured neuroblastoma cells (Omura et al, J. Antibiotics 44: ⁇ ⁇ 1 (1991); Omura et al, J. Antibiotics 44:113 (1991); Fenteany et al, Proc. Natl Acad. Sci. (USA) 91:335% (1994)).
  • the cellular target mediating these effects is the 20S proteasome, the proteolytic core of the 26S proteasome, which is the central component of the ubiquitin-proteasome pathway for intracellular protein degradation.
  • lactacystin inhibits the proteasome through the intermediacy of the active species, c/ ⁇ sto-lactacystin ⁇ -lactone (2), which specifically acylates the N-terminal threonine residue of the proteasome X/MBl subunit (Fenteany, et al, Science 268:726 (1995); Dick et al, J. Biol. Chem. 271:7273 (1996)). Lactacystin analogs are disclosed by Fenteany et al (WO 96/32105).
  • the ubiquitin-proteasome pathway is involved in a variety of important physiological processes (Goldberg et al, Chemistry & Biology 2:503 (1995); Ciechanover Cell 79:13 (1994); Deshaies, Trends Cell Biol.5:43 ⁇ (1995)). In fact, the bulk of cellular proteins are hydrolyzed by this pathway. Protein substrates are first marked for degradation by covalent conjugation to multiple molecules of a small protein, ubiquitin. The resultant polyubiquitinated protein is then recognized and degraded by the 26S proteasome. Long recognized for its role in degradation of damaged or mutated intracellular proteins, this pathway is now also known to be responsible for selective degradation of various regulatory proteins.
  • ubiquitin-proteasome pathway also mediates degradation of a number of other cell cycle regulatory proteins and tumor suppressor proteins (e.g., p21, p27, p53).
  • Activation of the transcription factor NF- ⁇ B which plays a central role in the regulation of genes involved in the immune and inflammatory responses, is dependent upon ubiquitination and degradation of an inhibitory protein, I ⁇ B- ⁇ (Palombella et al , " WO 95/25533).
  • I ⁇ B- ⁇ inhibitory protein
  • a first aspect of the present invention relates to a process for forming lactacystin or analogs thereof having Formula VI or c/ ⁇ st ⁇ -lactacystin ⁇ -lactone or analogs thereof having Formula VII:
  • R 1 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 2 is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxy alkyl, or amido, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 7 is alkyl, aryl, aralkyl, alkaryl, wherein any of said alkyl, aryl, aralkyl or alkaryl can be optionally substituted.
  • a second aspect of the present invention is directed to a method of forming formyl amides of Formula XIV:
  • R 2 is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amido, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 5 and R 6 are independently one of alkyl or alkaryl; or R 5 and R 6 when taken together with the nitrogen atom to which they are attached form a 5- to 7- membered heterocyclic ring, which may be optionally substituted, and which optionally may include an additional oxygen or nitrogen atom.
  • a third aspect of the present invention relates to forming tri-substituted oxazolines of Formula Ia or lb:
  • R 1 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted; and R 4 is aryl or heteroaryl, either of which may be optionally substituted.
  • the tri- substituted oxazolines of Formulae Ia and lb are useful as starting materials in forming lactacysin, c/ ⁇ sto-lactacystin ⁇ -lactone or analogs thereof via the process described herein.
  • a fourth aspect of the present invention is directed to lactacysin, clasto- lactacystin ⁇ -lactone or analogs of Formulae /and FiZ that possess unexpected biological activity.
  • Lactacystin, c/ ⁇ sto-lactacystin ⁇ -lactone, and analogs thereof possess biological activity as inhibitors of the proteasome. They can be used to treat conditions mediated directly by the function of the proteasome, such as muscle wasting, or mediated indirectly via proteins which are processed by the proteasome, such as the transcription factor NF- ⁇ B.
  • a fifth aspect of the present invention relates to pharmaceutical compositions, comprising a compound of Formula VI or Formula VII, and a pharmaceutically acceptable carrier or diluent.
  • a sixth aspect of the present invention relates to methods of inhibiting proteasome function or treating a condition that is mediated directly or indirectly by the function of the proteasome, by administering a compound of Formula VI or Formula VII that possesses unexpectedly high activity in inhibiting the proteasome.
  • Preferred Embodiments are directed to the use of a compound of Formulae VI or VII to prevent or reduce the size of infarct after vascular occlusion for example, for treating neuronal loss following stroke.
  • An additional preferred embodiment is directed to the use of said compounds for treating asthma.
  • a seventh aspect of the invention relates to enantiomerically-enriched compositions of formyl amides of Formula XIV.
  • An eighth aspect of the present invention relates to novel individual intermediates, such as aldols of Formula II and aminodiols of Formula III:
  • a ninth aspect of the present invention relates to individual intermediates, such as compounds of Formulae XVII, XVIII and XIX:
  • X is a halogen, preferably Cl, Br or I, as well as individual steps within the multistep process for forming substituted oxazolines of Formula /.
  • the present invention relates to an improved multi-step synthesis of lactacystin, c/ ⁇ sto-lactacystin ⁇ -lactone, and analogs thereof, that proceeds in fewer steps and in much greater overall yield than syntheses described in the prior art.
  • a number of individual process steps and chemical intermediates distinguish this synthetic pathway from pathways described in the prior art.
  • this synthetic pathway relies upon a novel stereospeciflc synthesis of an oxazoline intermediate, and a unique stereoselective addition of a formyl amide to the oxazoline.
  • the invention is also directed to novel analogs of Formulae VI and VII that possess unexpected biological activity.
  • Lactacystin, c/ sto-lactacystin ⁇ - lactone, and analogs thereof possess biological activity as inhibitors of the proteasome. They can be used to treat conditions mediated directly by the function of the proteasome, such as muscle wasting, or mediated indirectly via proteins which are processed by the proteasome, such as the transcription factor
  • the present invention is also directed to methods of inhibiting proteasome function or treating a condition that is mediated directly or indirectly by the function of the proteasome, by administering a compound of Formula VI or VII that possesses unexpectedly high activity in inhibiting the proteasome.
  • a pharmaceutical composition that includes a compound of Formula VI or Formula VII is administered to treat ischemic or reperfusion injury.
  • said compounds can be used to treat, prevent or ameliorate neuronal loss following stroke.
  • a first aspect of the present invention relates to processes for forming lactacystin and analogs thereof having Formula Viand c/ ⁇ sto-lactacystin ⁇ -lactone and analogs thereof having Formula VII:
  • R ! is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 2 is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amido, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted; and R 7 is alkyl, aryl, aralkyl, alkaryl, wherein any of said alkyl, aryl, aralkyl or alkaryl can be optionally substituted.
  • R 1 and R 2 are as defined above for Formulae VI and VII. These steps include:
  • R 1 is as defined above, and R 3 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, any of which can be optionally substituted;
  • R 4 is aryl or heteroaryl, either of which may be optionally substituted; by treating said substituted aryl or heteroary 1 oxazoline with a strong base to form an enolate;
  • R 2 is as defined above for Formulae VI and VII, and
  • R 5 and R 6 are independently one of alkyl or alkaryl; or R 5 and R 6 when taken together with the nitrogen atom to which they are attached form a 5- to 7- membered heterocyclic ring, which may be optionally substituted, and which optionally may include an additional oxygen or nitrogen atom, to form an adduct of Formula II:
  • R 1 and R 2 are as defined above.
  • the carboxylic acid intermediate of Formula V can be cyclized by treatment with a cyclizing reagent to form a c/ ⁇ t -lactacystin- ⁇ -lactone or analog thereof of Formula VII, which can be optionally further reacted with a thiol
  • R 7 SH such as N-acetylcysteine
  • the carboxylic acid intermediate of Formula K can be directly coupled to a thiol (R 7 SH), such as N-acetylcysteine, to form lactacystin or an analog thereof having Formula VI.
  • a thiol such as N-acetylcysteine
  • a second aspect of the present invention relates to the formation of enantiomerically-enriched formyl amides of Formula XIV:
  • R 2 and R 8 are as defined above;
  • R 2 , R 5 and R 6 are as defined above;
  • R 2 , R 5 and R 6 are as defined above;
  • a third aspect of the invention relates to a process for forming a tri- substituted cis-oxazoline compound of Formula la:
  • R 1 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted
  • R 3 is alkyl, cycloalkyl, aryl, alkaryl, any of which can be optionally substituted
  • R 4 is aryl or heteroaryl, either of which may be optionally substituted; said method comprising: (a) asymmetrically dihydroxylating an alkene intermediate of
  • X is a halogen, preferably Cl, Br or I;
  • the third aspect of the invention relates to a process for forming a tri-substituted trans-oxazoline compound of Formula lb comprising: (a) asymmetrically dihydroxylating an alkene intermediate of
  • X is a halogen, preferably Cl, Br, or I;
  • R 1 are C,. 12 alkyl, especially C,_ 8 alkyl, C 3.8 cycloalkyl, especially C 3.6 cycloalkyl, C 2 . 8 alkenyl, C 2 . 8 alkynyl, C 6. , 4 aryl, especially C 6.10 aryl, C 6.10 ar(C, .6 )alkyl or C ] . 6 alk(C 6 ., 0 )aryl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted.
  • Substituents that can be optionally present on the aryl ring of an R 1 moiety include one or more, preferably one or two, of hydroxy, nitro, trifluoromethyl, halogen, C, .6 alkyl, C 6.10 aryl, C,_ 6 alkoxy, C 1-6 aminoalkyl, C,. 6 aminoalkoxy, amino, C 2 _ 6 alkoxycarbonyl, carboxy, C,_ 6 hydroxyalkyl, C 2 . 6 hydroxyalkoxy, C,_ 6 alkylsulfonyl, C 6 ., 0 arylsulfonyl, C, .6 alkylsulfinyl, C,.
  • R 1 is more preferably one of C,. g alkyl such as ethyl, propyl or isopropyl; cycloalkyl, such as cyclohexyl; or C 6 . 10 aryl, such as phenyl. Most preferred is isopropyl.
  • Preferred values of R 2 are C,. g alkyl, C 3 . g cycloalkyl, especially C 3.6 cycloalkyl, C,. g alkoxy, C 2 . g alkenyl, C 2 . g alkynyl C 6.14 aryl, especially C 6.10 aryl, C 6 . ]0 ar(C,. 6 )alkyl or C,. 6 alk(C 6 . ]0 )aryl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted with any of the substituents as described for R 1 above.
  • R 2 is more preferably C M alkyl, such as methyl, ethyl, propyl, or butyl; or
  • C M alkoxy such as methoxy, or ethoxy. Most preferred are methyl, ethyl and propyl, and butyl.
  • ester functionalities can be employed at this position.
  • Preferred values are C,. g alkyl, C 3 . 8 cycloalkyl, especially C 4.7 cycloalkyl,
  • R 3 C, .6 alk(C 6.10 )aryl, any of which can be optionally substituted.
  • Substituents that can be optionally present on R 3 include one or more, preferably one or two, of the substituents as described for R 1 above.
  • R 3 is more preferably C alkyl, C 6 . I0 aryl or C 6 . I0 ar(C,. 6 )alkyl. Most preferred are methyl, ethyl, tert-butyl and benzyl.
  • R 4 is preferably C 6 . 10 aryl, preferably phenyl, or a heteroaryl group selected from the group consisting of thienyl, benzo[b]thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, or triazolyl.
  • the phenyl or heteroaryl group can be optionally substituted by one or two of the substituents as described for R 1 above. Most preferred are phenyl, and phenyl substituted by halogen, C,. 6 alkyl, C,. 6 alkoxy, carboxy, amino, C,_ 6 alkylamino and or di(C,. 6 )alkylamino.
  • R 5 and R 6 are independently one of alkyl, aralkyl or alkaryl; or R 5 and R 6 when taken together with the nitrogen atom to which they are attached form a 5- to 7-membered heterocyclic ring, which can be optionally substituted, and which optionally can include an additional oxygen or nitrogen atom.
  • Optional substituents are those listed above for R 1 .
  • R 5 and R 6 are preferably C,. 6 alkyl, C 6 . ]0 ar(C, .6 )alkyl or C, .6 alk(C 6. ⁇ 0 )aryl or together with the nitrogen atom to which they are attached form a 5- to 7- membered heterocycle which can be optionally substituted, and which optionally can include an additional oxygen or nitrogen atom.
  • Most preferred values for NR 5 R 6 are dimethylamino, diethylamino, pyrrolidino, piperidino, morpholino, oxazolidinone, and oxazolidinone substituted by halogen, C,_ 6 alkyl, C 6 . 10 ar(C, .6 )alkyl, C,.
  • R 7 is preferably C,. g alkyl, C 3 . 8 cycloalkyl, C 6 . 10 aryl, C 6.10 ar(C, .6 )alkyl,
  • R 7 C, .6 alk(C 6.]0 )aryl, any of which can be optionally substituted.
  • Substituents that can be optionally present on either or both of the ring or chain portions of R 7 include one or more, preferably one or two, of the substituents as described for R 1 above.
  • R 7 together with the sulfur atom to which it is attached is cysteine or a derivative of cysteine such as N-acetyl cysteine, glutathione, and the like.
  • Scheme 1 is a general scheme for forming lactacystin and c/ ⁇ st ⁇ -lactacystin- ⁇ -lactone analogs from substituted oxazoline starting materials.
  • the starting oxazoline / which may be of either the cis (Ia) or trans (lb) configuration, is deprotonated with a strong base to form the enolate.
  • bases suitable for use in this reaction are organic bases, including hindered amide bases such as lithium diisopropylamide (LDA), lithium tetramethylpiperidide (LiTMP), lithium, sodium or potassium hexamethyldisilazide (LiHMDS,
  • reaction temperatures preferably range from about -100°C to about -30°C, more preferably from -85 °C to -50°C, and most preferably from -85 °C to -75 °C.
  • the reaction temperature is important in determining the stereochemical outcome of the subsequent addition to the aldehyde, with lower temperatures providing better selectivity.
  • the deprotonation step is followed by transmetallating said enolate with a metal selected from the group consisting of titanium, aluminum, tin, zinc, magnesium and boron.
  • Preferred reagents for this step include titanium or aluminum Lewis acids, for example Me 2 AlCl or ( -PrO) 3 TiCl or a mixture of the two.
  • a formyl amide (XIV) affords the adduct//.
  • Useful catalysts for this reaction include palladium black, palladium on activated carbon, palladium hydroxide on carbon, or the like.
  • Organic solvents suitable for use in this reaction include lower alkanols such as methanol, ethanol, or isopropanol, lower alkanoates such as ethyl acetate, lower alkanoic acids such as acetic acid, or mixtures thereof.
  • the reaction is conducted under an atmosphere of hydrogen, at pressures ranging from about 15 to about 100 p.s.i., more preferably from about 30 to about 50 p.s.i.
  • transfer hydrogenation procedures R.A.W. Johnstone et al, Chem. Rev. 55:129 (1985)
  • the adduct // is treated at atmospheric pressure with a catalyst and a hydrogen donor.
  • the aminodiol /// is converted to the ⁇ -lactam IV, which can then be isolated in approximately 60-75% overall yield from//.
  • the heating step is conveniently carried out by first filtering off the catalyst used in the hydrogenation step and then heating the filtrate to reflux.
  • Isopropenyl chloroformate is a preferred reagent for this step, since all byproducts are volatile and chromatographic purification of the product is not necessary.
  • c/ wto-Lactacystin ⁇ -lactone can be converted to lactacystin by treatment of the ⁇ -lactone with N-acetylcysteine according to the reported procedure (Corey et al, Tetrahedron Lett. 34:6977 (1993)). Reactions of the ⁇ -lactone //with other thiols proceed analogously.
  • lactacystin analogs are prepared by coupling the carboxylic acid intermediate V with a thiol to form the corresponding thiolester VI. The method of this invention is therefore useful for synthesis of both lactacystin and c/ ⁇ sto-lactacystin ⁇ -lactone, as well as analogs thereof.
  • the enantiomerically-enriched formyl amides XIV employed in the aldol reaction are new. They can be prepared according to a representative reaction sequence such as that depicted in Scheme 2.
  • the term "enantiomerically-enriched” means that one enantiomer is present in excess relative to the other; that is, one enantiomer represents greater than 50% of the mixture.
  • stereoselective is used to mean that a synthesis or reaction step produces one enantiomer or diastereomer in excess relative to the other enantiomer or to other diastereomer(s).
  • Peroxide mediated hydrolysis affords the acid XI, which is coupled with an amine to provide the amide XII, generally in greater than 50% overall yield.
  • Benzyl group hydrogenolysis followed by oxidation of the resultant alcohol (XIII) then affords the formyl amide XlVin 80-85% yield.
  • Pearlmans catalyst Pd(OH) 2
  • the final oxidation step is best accomplished with the periodinane reported by Dess and Martin, J. Org. Chem. 48:4156 (1983) or with 2,2,6,6-tetramethyl-l-piperidinyloxy (TEMPO) free radical, and buffered hypochlorite in the presence of bromide ion (J.
  • XIV can be shown to be enantiomerically pure by reducing the aldehyde with sodium borohydride and converting the resultant alcohol to the corresponding Mosher ester using 7?-(+)- ⁇ -methoxy- ⁇ -(trifluoromethyl) phenylacetyl chloride (Dale et al, J. Org. Chem. 34:2543 (1969)).
  • ⁇ NMR analysis at 300 MHz reveals a single diastereomer.
  • the aldehydes prepared according to Scheme 2 are configurationally stable, showing no signs of enantiomeric deterioration after one week, when stored at 0 °C.
  • the aldehyde is also configurationally stable under the conditions of the aldol reaction, and the adduct // is formed without epimerization of the substituent R 2 at C(7).
  • the synthetic methods will work with any substituent at R 1 that is stable to strong base and to hydrogenation. Isopropyl is the preferred substituent for good proteasome inhibiting activity of the final product.
  • the invention also relates to a new route to form the oxazoline starting material /.
  • the overall synthesis includes five steps (Scheme 3) and affords the c/5-substituted oxazoline Ia, which is thereafter employed in the method described above.
  • the first step depicted in Scheme 3 is Sharpless asymmetric dihydroxylation (Sharpless et al, J. Org. Chem. 57:2768 (1992); Kolb et al, Chem. Rev. 94:2483 (1994); Shao and Goodman, J. Org. Chem. (57:2582 (1996)) of the alkene XV.
  • the alkene XV is prepared by Wittig condensation between the aldehyde and carboethoxymethylene triphenylphosphorane (Hale et al, Tetrahedron 50:9181 (1994)). Other olefination procedures are also known in the art.
  • the dihydroxylation reaction is preferably conducted with AD-mix- ⁇ (Aldrich Chemical Co.) in the presence of methane sulfonamide and stereoselectively affords the diol XVIa, as predicted by the Sharpless face-selection rule.
  • the dihydroxylation reaction is preferably conducted using N-methylmorpholine-N-oxide (NMO) as the reoxidant in place of K 3 Fe(CN) 6 present in AD-mix- ⁇ .
  • NMO N-methylmorpholine-N-oxide
  • this procedure allows more concentrated reaction mixtures and greatly simplifies the workup.
  • the enantiomeric purity of the product can be enhanced by recrystallization.
  • the diol XVIa is treated with an orthoester under Lewis or Br ⁇ nsted acid catalysis to give a mixed orthoester, which is converted in situ to the haloester XVIIa by treatment with an acyl halide (Haddad et al, Tetrahedron Lett.
  • acyl halides especially acetyl halides are preferred for this reaction, other acid halides such as HCl, HBr, HI, Me 3 SiCl, Me 3 SiI, Me 3 SiBr and the like may be used.
  • Halogen-containing Lewis acids of the formula ML n X such as BBr 3 , SnCl 4 , Ti(OR) 2 Cl 2 , Ti(OR) 3 Cl, and the like can also be used.
  • M is a metal selected from the group consisting of B, Ti, Sn, Al, Zn, and Mg; L is any suitable ligand for the metal, preferably an alkoxide or halogen group; n is an integer that results in a stable complex, and X is a halogen.
  • acetyl bromide is used to produce the haloester XVIIa.
  • the orthoester employed in this reaction is derived from an aromatic or heteroaromatic carboxylic acid. More preferably, the orthoester is derived from benzoic acid, e.g., trimethyl orthobenzoate.
  • boron trifluoride etherate as the Lewis acid catalyst in the formation of the mixed orthoester is preferred, but other acids, such as HBr, SnCl 4 , TiCl 4 , BBr 3 , and the like, can also be used.
  • the crude halide XVIIa is converted to the azide XVIIIa by treatment with an alkali metal azide in a polar aprotic organic solvent, such as dimethyl sulfoxide (DMSO) or N,N-dimethyl formamide (DMF).
  • a polar aprotic organic solvent such as dimethyl sulfoxide (DMSO) or N,N-dimethyl formamide (DMF).
  • DMSO dimethyl sulfoxide
  • DMF N,N-dimethyl formamide
  • Treatment of XlXa with thionyl chloride in methylene chloride effects ring closure with inversion of configuration at the hydroxyl-substituted carbon atom to produce the cw-substituted oxazoline starting material Ia.
  • Other reagents suitable for use in this reaction include sulfuryl chloride, phosphorous trichloride, phosphorous oxychloride, and (methoxycarbonylsulfamoyl)-triethylammonium hydroxide, inner salt (Burgess reagent).
  • Treatment of XlXa under Mitsunobu conditions (Mitsunobu, Synthesis:! (1981) will also effect a ring closure.
  • the oxazoline ring oxygen atom is destined to become the C(9)-hydroxyl group in the final products VI and VII.
  • the cw-oxazoline (Ia) is converted to the trflr ⁇ -oxazoline (lb) by inversion of configuration of the ester substituent, with the configuration of the
  • fifth and sixth aspects of the invention relate to lactacystin analogs that can be made by the synthetic routes described herein; to pharmaceutical compositions including such compounds; and to methods of treating a subject having a condition mediated by proteins processed by the proteasome by administering to a subject an effective amount of a pharmaceutical composition disclosed herein.
  • These methods include treatments for Alzheimers disease, cachexia, cancer, inflammation (e.g., inflammatory responses associated with allergies, bone marrow or solid organ transplantation, or disease states, including but not limited to arthritis, multiple sclerosis, inflammatory bowel disease and parasitic diseases such as malaria), psoriasis, restenosis, stroke, and myocardial infarction.
  • the compounds of Formulae VI and VII disclosed herein are highly selective for the proteasome, and do not inhibit other proteases such as trypsin, oc-chymotrypsin, calpain I, calpain II, papain, and cathepsin B.
  • lactacystin, c/ ⁇ sto-lactacystin ⁇ -lactone, and analogs thereof possess biological activity as inhibitors of the proteasome. They can be used to treat conditions mediated directly by the function of the proteasome, such as muscle wasting, or mediated indirectly via proteins which are processed by the proteasome, such as the transcription factor NF- ⁇ B.
  • the compounds prepared by the methods of this invention can also be used to determine whether a cellular, developmental, or physiological process or output is regulated by the proteolytic activity of the proteasome.
  • R 1 is C, .12 alkyl, C 3 . g cycloalkyl, C 2 . 8 alkenyl, C 2 . g alkynyl, C 6 . 14 aryl, C 6 . 10 ar(C, .6 )alkyl or C 1 . 6 alk(C 6 . 10 )aryl;
  • R 2 is C 2 . 6 alkyl; and R 7 is C,. g alkyl, C 3 . g cycloalkyl, C 6 . 10 aryl, C 6.10 ar(C, .6 )alkyl,
  • R 1 is C, .4 alkyl, more preferably isopropyl.
  • R 2 is preferably ethyl, n-propyl, n-butyl or isobutyl.
  • R 7 together with the sulfur atom to which it is attached is cysteine or a derivative of cysteine such as N-acetyl cysteine, glutathione, and the like.
  • a seventh aspect of the present invention is directed to enantiomerically- enriched formyl amides of Formula XIV:
  • R 2 is C,. g alkyl, C 3 . g cycloalkyl, C 2 . 8 alkenyl, C 2 . 8 alkynyl, C 6.14 aryl, C 6.10 ar(C,. 6 )alkyl or C,. 6 alk(C 6 . ]0 )aryl; and
  • R 5 and R 6 are independently C,. 6 alkyl, C 6 ., 0 ar(C,. 6 )alkyl or C 1.6 alk(C 6.10 )aryl, or together with the nitrogen atom to which they are attached form a 5- to 7-membered heterocycle which can be optionally substituted, and which optionally can include an additional oxygen or nitrogen atom.
  • Preferred compounds are those where R 2 is C 2.6 alkyl.
  • An eighth aspect of the present invention is directed to compounds of Formulae // and ///:
  • R 1 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 2 is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amido, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 3 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, any of which can be optionally substituted;
  • R 4 is optionally substituted aryl or optionally substituted heteroaryl
  • R 5 and R 6 are independently one of alkyl or alkaryl; or R 5 and R 6 when taken together with the nitrogen atom to which they are attached form a 5- to 7- membered heterocyclic ring, which can be optionally substituted, and which optionally include an additional oxygen or nitrogen atom.
  • Most preferred values for NR 5 R 6 are dimethylamino, diethylamino, pyrrolidino, piperidino, morpholino, oxazolidinone, and oxazolidinone substituted by halogen, C,. 6 alkyl, C 6 . 10 ar(C, .6 )alkyl, C ⁇ alkoxy, carboxy, and/or amino.
  • Preferred compounds of Formulae // and /// are those wherein: R 1 is C,. 12 alkyl, C 3 . g cycloalkyl, C 2 . 8 alkenyl, C 2.8 alkynyl, C 6.14 aryl, C 6 . ]0 ar (C j . ⁇ alkyl or C,. 6 alk(C 6 . 10 )aryl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted; R 2 is C,. g alkyl, C 3 . 8 cycloalkyl, C 2 .
  • alkenyl C 2.g alkynyl, C 6.14 aryl, C 6.10 ar(C, .6 )alkyl or C,. 6 alk(C 6.10 )aryl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 3 is C,. g alkyl, C 3 . g cycloalkyl, C 2 . g alkenyl, C 2 . g alkynyl, C 6.I4 aryl, C 6 ., 0 ar(C, .6 )alkyl or C,. 6 alk(C 6 . 10 )aryl, any of which can be optionally substituted;
  • R 4 is optionally substituted C 6.10 aryl, or an optionally substituted heteroaryl group selected from the group consisting of thienyl, benzo[ ⁇ ]thienyl, fiiryl, pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinoliziny 1, isoquinolyl, quinolyl, or triazolyl; and
  • R 5 and R 6 are independently C, .6 alkyl, C 6. , 0 ar(C, .6 )alkyl or C, .6 alk(C 6.]0 )aryl, or together with the nitrogen atom to which they are attached form a 5- to 7-membered heterocycle which can be optionally substituted, and which optionally can include an additional oxygen or nitrogen atom.
  • Most preferred values for NR 5 R 6 are dimethyl ami no, diethylamino, pyrrolidino, piperidino, morpholino, oxazolidinone, and oxazolidinone substituted by halogen, C,. 6 alkyl, C 6 . ]0 ar(C,. 6 )alkyl, C,_ 6 alkoxy, carboxy, and/or amino.
  • a ninth aspect of the present invention is directed to compounds of Formulae XVIIa, XVIIb, XVIIIa, XVIIIb, XlXa or XlXb:
  • R 1 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, aralkyl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 3 is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, any of which can be optionally substituted;
  • R 4 is optionally substituted aryl or optionally substituted heteroaryl.
  • Preferred compounds of Formulae XVII, XVIII or XIX are those wherein
  • R 1 is C 2 alkyl, C 3 . g cycloalkyl, C 2 . 8 alkenyl, C 2 . 8 alkynyl C 6 . 14 aryl, C 6 . 10 ar(C, .6 )alkyl or C,_ 6 alk(C 6.10 )aryl, where the ring portion of any of said aryl, aralkyl, or alkaryl can be optionally substituted;
  • R 3 is C, .8 alkyl, C 3.g cycloalkyl, C 2 . 8 alkenyl, C 2.g alkynyl, C 6.I4 aryl, C, 10 ar(C,. 6 )alkyl or C, .6 alk(C 6 .
  • R 4 is optionally substituted C 6 ., 0 aryl, or an optionally substituted heteroaryl group selected from the group consisting of thienyl, benzo[b]thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, or triazolyl.
  • alkyl as employed herein includes both straight and branched chain radicals of up to 12 carbons, preferably 1 -8 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, 1-ethylpropyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl and dodecyl.
  • substituted alkyl includes alkyl groups as defined above that have one, two or three halo, hydroxy, nitro, trifluoromefhyl, halogen, C,. 6 alkyl, C 6 . 10 aryl, C,. 6 alkoxy, C,_ 6 aminoalkyl, C, .6 aminoalkoxy, amino, C 2 . 6 alkoxycarbonyl, carboxy, C,. 6 hydroxyalkyl, C 2.6 hydroxyalkoxy, C, .6 alkylsulfonyl, C 6 . 10 arylsulfonyl, C,_ 6 alkylsulfinyl, C,.
  • cycloalkyl as employed herein includes saturated cyclic hydrocarbon groups containing 3 to 12 carbons, preferably 3 to 8 carbons, which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, any of which groups may be substituted with substituents such as halogen, C,. 6 alkyl, C, .6 alkoxy and/or hydroxy group.
  • heteroaryl refers to groups having 5 to 14 ring atoms, preferably 5, 6, 9 or 10 ring atoms; 6, 10 or 14 ⁇ electrons shared in a cyclic array; and containing carbon atoms and 1 , 2 or 3 oxygen, nitrogen or sulfur heteroatoms (where examples of heteroaryl groups are: thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indo
  • aryl as employed herein by itself or as part of another group refers to monocyclic or bicyclic aromatic groups containing from 6 to 12 carbons in the ring portion, preferably 6-10 carbons in the ring portion, such as phenyl, naphthyl or tetrahydronaphthyl.
  • aralkyl or “arylalkyl” as employed herein by itself or as part of another group refers to C,. 6 alkyl groups as discussed above having an aryl substituent, such as benzyl, phenylethyl or 2-naphthylmethyl.
  • alkaryl or “alkylaryl” as employed herein by itself or as part of another group refers to an aryl group as discussed above having a C,_ 6 alkyl substituent, such as toluyl, ethylphenyl, or methylnaphthyl.
  • aryl, aralkyl, alkaryl or 5-, 6-, 9- or 10- membered heteroaryl groups means that the ring portion of said groups can be optionally substituted by one or two substituents independently selected from C,_ 6 alkyl, C 3 . 8 cycloalkyl, C,_ 6 alkyl(C 3 . 8 )cycloalkyl, C 2.g alkenyl, C 2 . g alkynyl, cyano, amino, C,_ 6 alkylamino, di(C,.
  • alkoxy refers to the above alkyl groups linked to oxygen.
  • halogen or "halo" as employed herein by itself or as part of another group refers to chlorine, bromine, fluorine or iodine.
  • amido refers to formylamino, alkylcarbonylamino or arylcarbonylamino.
  • the lactone ring is subject to nucleophilic attack not only by the threonine residue of the proteasome X/MBl subunit, but also by water. Hydrolysis results in formation of the hydroxy acid V, which is not active as an inhibitor of the proteasome.
  • Relative potency in cell culture is a composite of many factors, including enzyme potency, cell penetration, and hydrolysis rate. Although more potent than 2 against the enzyme, 3f is also more rapidly hydrolyzed, resulting in much weaker activity in cell culture. By contrast, the analogs 3a-3d show unexpectedly improved potency not only in the enzyme assay, but also in cell culture.
  • the disclosed compounds are used to treat conditions mediated directly by the proteolytic function of the proteasome such as muscle wasting, or mediated indirectly via proteins which are processed by the proteasome such as ⁇ F- ⁇ B.
  • the proteasome participates in the rapid elimination and post-translational processing of proteins involved in cellular regulation (e.g., cell cycle, gene transcription, and metabolic pathways), intercellular communication, and the immune response (e.g., antigen presentation).
  • Specific examples include ⁇ -amyloid protein and regulatory proteins such as cyclins and transcription factor NF- ⁇ B.
  • Treating as used herein includes reversing, reducing, or arresting the symptoms, clinical signs, and underlying pathology of a condition in a manner to improve or stabilize the subject's condition.
  • proteasome inhibitors are useful for treating conditions such as cancer, chronic infectious diseases, fever, muscle disuse (atrophy) and denervation, nerve injury, fasting, renal failure associated with acidosis, and hepatic failure. See, e.g., Goldberg, U.S. Pat. No. 5,340,736 (1994).
  • Embodiments of the invention therefore encompass methods for reducing the rate of muscle protein degradation in a cell, and reducing the rate of intracellular protein degradation.
  • Each of these methods includes the step of contacting a cell (in vivo or in vitro, e.g., a muscle in a subject) with an effective amount of a compound (e.g., pharmaceutical composition) of a formula disclosed herein.
  • Proteasome inhibitors block processing of ubiquitinated NF- ⁇ B in vitro and in vivo. Proteasome inhibitors also block I ⁇ B- ⁇ degradation and NF- ⁇ B activation. (Palombella, et al; and Traenckner, et al, EMBO J. 73:5433-5441 (1994)).
  • One embodiment of the invention is a method for inhibiting I ⁇ B- ⁇ degradation, including contacting the cell with a compound of a formula described herein.
  • a further embodiment is a method for reducing the cellular content of NF- KB in a cell, muscle, organ, or subject, including contacting the cell, muscle, organ, or subject with a compound of a formula described herein.
  • Additional embodiments encompass methods for treating inflammatory responses associated with allergies, bone marrow or solid organ transplantation, or disease states, including but not limited to arthritis, inflammatory bowel disease, asthma, and multiple sclerosis by administering a compound of a formula disclosed herein.
  • a preferred embodiment of the invention is directed to treating asthma by administering a compound of Formula VI or Formula VII, most preferably compound 3b.
  • Proteasome inhibitors are also useful for treatment of ischemic or reperfusion injury, particularly for preventing or reducing the size of infarct after vascular occlusion such as occurs during a stroke or heart attack, as described in
  • Proteasome inhibitors also block proteasome-dependent transformation of protazoan parasites (Gonzalez et al , J.
  • Said compounds can be administered from about 0 to about 10 hours after the occurrence of a stroke in order to treat or reduce neuronal loss following an ischemic event.
  • Compounds 3b is the most preferred compound in this aspect of the invention.
  • Proteasome inhibitors also block degradation of cell cycle regulatory proteins, such as cyclins and cyclin-dependent kinase inhibitors, and tumor suppressor proteins, such as p53.
  • Other embodiments of the invention therefore encompass methods for blocking the cell cycle and for treating cell proliferative diseases such as cancer, psoriasis, and restenosis with a compound of a formula described herein.
  • the term "inhibitor” is meant to describe a compound that blocks or reduces the activity of an enzyme (e.g., the proteasome, or the X/MB 1 subunit of the 20S proteasome).
  • An inhibitor may act with competitive, uncompetitive, or noncompetitive inhibition.
  • An inhibitor may bind reversibly or irreversibly, and therefore the term includes compounds which are suicide substrates of an enzyme.
  • An inhibitor may modify one or more sites on or near the active site of the enzyme, or it may cause a conformational change elsewhere on the enzyme.
  • Amounts and regimens for the administration of proteasome inhibitors and compositions of the invention can be determined readily by those with ordinary skill in the clinical art.
  • the dosage of the composition of the invention will vary depending upon considerations such as: type of composition employed; age; health; medical conditions being treated; kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired; extent of tissue damage; gender; duration of the symptoms; and, counter indications, if any, and other variables to be adjusted by the individual physician.
  • a desired dosage can be administered in one or more applications to obtain the desired results.
  • Pharmaceutical compositions containing the proteasome inhibitors of the invention can be provided in unit dosage forms.
  • compositions within the scope of this invention include all compositions wherein the compounds of the present invention are contained in an amount which is effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art.
  • the compounds may be administered to mammals, e.g. humans, orally at a dose of 0.0025 to 50 mg/kg, or an equivalent amount of the pharmaceutically acceptable salt thereof, per day of the body weight of the mammal being treated for a proteosome-mediated condition such as a stroke or asthma.
  • the dose is generally about one-half of the oral dose.
  • the compound in the method of prevention or reduction of infarct size can be administered by intravenous injection at a dose of about 0.01 to about 10 mg/kg, preferably about 0.025 to about 1 mg/kg.
  • the unit oral dose may comprise from about 0.01 to about 50 mg, preferably about 0.1 to about 10 mg of the compound.
  • the unit dose may be administered one or more times daily as one or more tablets each containing from about 0.1 to about 10, conveniently about 0.25 to 50 mg of the compound or its solvates.
  • a single dosage be administered, 0 to about 10 hours post-event, preferably 0 to about 6 hours post- event.
  • 4- Methylvaleryl chloride was prepared from commercially available 4-methylvaleric acid in the following way : a cold (0 ° C) solution of 4-methy Ivaleric acid (1.85 mL, 15.0 mmol) in 50 mL anhydrous CH 2 C1 2 containing 10 mL of DMF was treated with 1.95 ⁇ L oxalyl chloride (22.5 mmol). The mixture was then stirred for 3 h at room temperature, concentrated in vacuo and filtered to affords 1.65 g (100%>) of the desired acid chloride as a colorless liquid. ii.
  • aqueous layer was extracted with AcOEt (2 x 20 mL) and the combined organic layers were washed successively with 0.5 N aqueous HCl (20 mL), H 2 O (20 mL), 0.5 M aqueous NaHSO 3 (2 x 15 mL), saturated aqueous NaHCO 3 and finally with brine, then dried over Na 2 SO 4 and concentrated in vacuo affording 879 mg (> 100%) of crude Aldol product lib which was pure enough to be used directly in the subsequent step.
  • Aldol product lib was also obtained in 100% yield by a procedure analogous to that described above but using cis-oxazoline lb (see below) instead of tr ⁇ s-oxazoline Ia.
  • reaction mixture was purged with nitrogen and additional Pd(OH) 2 /C (1.3 g) was added.
  • the reaction mixture was purged with hydrogen and again purged every hour for 4 h.
  • the mixture was filtered and concentrated in vacuo.
  • the residue was dissolved in water and extracted with EtOAc.
  • the aqueous layer was basified with Na 2 CO 3 and again extracted with EtOAc.
  • the combined organic extracts were washed with brine, dried over Na 2 SO 4 , and concentrated to give a mixture of N- and O-benzoylated products, which was used directly in the next step. f.
  • C2C12 cells (a mouse myoblast line) were labeled for 48 hrs with 35 S- methionine. The cells were then washed and preincubated for 2 hrs in the same media supplemented with 2 mM unlabelled methionine. The media was removed and replaced with a fresh aliquot of the preincubation media containing 50%) serum, and a concentration of the compound to be tested. The media was then removed and made up to 10% TCA and centrifuged. The TCA soluble radioactivity was counted. Inhibition of proteolysis was calculated as the percent decrease in TCA soluble radioactivity. From this data, an IC 50 for each compound was calculated.
  • Example 14 Lactone Hydrolysis
  • mice Male Sprague Dawley rats (250-400 g) were anesthetized with haloethane and subjected to middle cerebral artery (MCA) occlusion using a nylon filament for 2 h. Subsequently, the filament was removed and reperfusion of the infarcted tissue occurred for 24 hours before the rat was sacrificed. Immediately after the filament was withdrawn, the animals were evaluated using a neurological scoring system. Neurological scores were expressed on a scale from 0 to 10, with 0 representing no neurological deficit and 10 representing severe neurological deficit. After 24 hours and before sacrifice, animals were evaluated a second time using the same neurological scoring system.
  • TTC triphenyltetrazolium chloride
  • Two additional groups of rats were given i.v. bolus injections (1.0 mL/kg) of 3b at 0 minutes, 2 hours, and 6 hours after the start of the occlusion.
  • infarct volume was decreased by 50% (FIG. 1, 0.3 x 1). Infarct volume was not significantly decreased in either the 0.1 mg/kg x 3 dosage group or the 0.3 mg/kg x 3 dosage group (FIG. 1).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Vascular Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Epidemiology (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Oncology (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Pyrrole Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)

Abstract

La présente invention a trait à une synthèse améliorée de clasto-lactacystine-β-lactone et de ses analogues, dont la mise en oeuvre comporte moins d'étapes et permet d'obtenir des rendements globalement beaucoup plus élevés que les synthèses décrites dans les techniques actuelles. La voie de synthèse repose sur une nouvelle synthèse stéréospécifique d'un produit intermédiaire d'oxazoline, et sur une addition stéréosélective unique d'un amide formyle à l'oxazoline. L'invention concerne également de nouvelles clasto-lactacystine-β-lactones et leurs analogues, ainsi que l'utilisation de ces composés comme inhibiteurs de protéasomes.
PCT/US1998/016858 1997-08-15 1998-08-14 SYNTHESE DE CLASTO-LACTACYSTINE β-LACTONE ET DE SES ANALOGUES WO1999009006A1 (fr)

Priority Applications (8)

Application Number Priority Date Filing Date Title
AU89062/98A AU749857B2 (en) 1997-08-15 1998-08-14 Synthesis of clasto-lactacystin beta-lactone and analogs thereof
KR1020007001558A KR20010022950A (ko) 1997-08-15 1998-08-14 클라스토-락타시스틴 β-락톤 및 이들의 유사체를 합성하는 방법
NZ503169A NZ503169A (en) 1997-08-15 1998-08-14 Synthesis of clasto-lactacystin beta-lactone whereupon the process relies upon a stereoselective addition of a formyl amide to an oxazoline
BR9811304-6A BR9811304A (pt) 1997-08-15 1998-08-14 Sìntese de clasto-lactacistina" beta"-lactona eseus análogos
CA002301054A CA2301054A1 (fr) 1997-08-15 1998-08-14 Synthese de clasto-lactacystine .beta.-lactone et de ses analogues
IL13453898A IL134538A (en) 1997-08-15 1998-08-14 Lactacystin derivatives and pharmaceutical compositions containing the same
JP2000509690A JP2001515064A (ja) 1997-08-15 1998-08-14 クラスト−ラクタシスチンβ−ラクトンおよびそのアナログの合成
EP98940885A EP1021407A4 (fr) 1997-08-15 1998-08-14 Synthese de clasto-lactacystine beta-lactone et de ses analogues

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US91211197A 1997-08-15 1997-08-15
US5584897P 1997-08-15 1997-08-15
US60/055,848 1997-08-15
US08/912,111 1997-08-15
US6735297P 1997-12-03 1997-12-03
US60/067,352 1997-12-03

Publications (1)

Publication Number Publication Date
WO1999009006A1 true WO1999009006A1 (fr) 1999-02-25

Family

ID=27368920

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1998/016858 WO1999009006A1 (fr) 1997-08-15 1998-08-14 SYNTHESE DE CLASTO-LACTACYSTINE β-LACTONE ET DE SES ANALOGUES

Country Status (11)

Country Link
EP (1) EP1021407A4 (fr)
JP (1) JP2001515064A (fr)
KR (1) KR20010022950A (fr)
CN (1) CN1271342A (fr)
AU (1) AU749857B2 (fr)
BR (1) BR9811304A (fr)
CA (1) CA2301054A1 (fr)
HU (1) HUP0002724A3 (fr)
IL (1) IL134538A (fr)
NZ (1) NZ503169A (fr)
WO (1) WO1999009006A1 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002082091A3 (fr) * 2001-04-09 2004-03-04 Allan Christian Shaw Procede d'identification de proteines a partir de bacteries intracellulaires
WO2004012732A3 (fr) * 2002-07-31 2004-04-29 Charite Universitaetsmedizin Utilisation d'un inhibiteur du proteasome dans le traitement du dysfonctionnement endothelial et/ou dans le cadre d'une therapie a base de proteasome a faible dose
US7144723B2 (en) 2000-11-16 2006-12-05 The Regents Of The University Of California Marine actinomycete taxon for drug and fermentation product discovery
US7176232B2 (en) 2002-06-24 2007-02-13 The Regents Of The University Of California Salinosporamides and methods for use thereof
US7179834B2 (en) 2002-06-24 2007-02-20 The Regents Of The University Of California Salinosporamides and methods for use thereof
US7276530B2 (en) 2004-04-30 2007-10-02 Nereus Pharmaceuticals, Inc. [3.2.0] Heterocyclic compounds and methods of using the same
US7579371B2 (en) 2004-04-30 2009-08-25 Nereus Pharmaceuticals, Inc. Methods of using [3.2.0] heterocyclic compounds and analogs thereof
US7824698B2 (en) 2007-02-02 2010-11-02 Nereus Pharmaceuticals, Inc. Lyophilized formulations of Salinosporamide A
US7842814B2 (en) 2006-04-06 2010-11-30 Nereus Pharmaceuticals, Inc. Total synthesis of salinosporamide A and analogs thereof
US7910616B2 (en) 2008-05-12 2011-03-22 Nereus Pharmaceuticals, Inc. Proteasome inhibitors
US8003802B2 (en) 2008-03-07 2011-08-23 Nereus Pharmaceuticals, Inc. Total synthesis of Salinosporamide A and analogs thereof
US8168803B2 (en) 2003-06-20 2012-05-01 Nereus Pharmaceuticals, Inc. Methods of using [3.2.0] heterocyclic compounds and analogs thereof
US8217072B2 (en) 2003-06-20 2012-07-10 The Regents Of The University Of California Salinosporamides and methods for use thereof
US8394816B2 (en) 2007-12-07 2013-03-12 Irene Ghobrial Methods of using [3.2.0] heterocyclic compounds and analogs thereof in treating Waldenstrom's Macroglobulinemia
US8722724B2 (en) 2004-12-03 2014-05-13 Triphase Research And Development I Corp. Compositions and methods for treating neoplastic diseases

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5831068A (en) * 1995-08-21 1998-11-03 Duke University Method to increase the density of antigen on antigen presenting cells

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5831068A (en) * 1995-08-21 1998-11-03 Duke University Method to increase the density of antigen on antigen presenting cells

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
COREY E. J., REICHARD G. A.: "TOTAL SYNTHESIS OF LACTACYSTIN.", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, AMERICAN CHEMICAL SOCIETY, US, vol. 114., 1 December 1992 (1992-12-01), US, pages 10677 - 10679., XP002915909, ISSN: 0002-7863, DOI: 10.1021/ja00052a096 *
See also references of EP1021407A4 *
SHIBATA K., ET AL.: "KINETIC AND THERMODYNAMIC CONTROL OF L-THREONINE ALDOLASE CATALYZED REACTION AND ITS APPLICATION TO THE SYNTHESIS OF MYCESTERICIN D.", TETRAHEDRON LETTERS, PERGAMON, GB, vol. 37., no. 16., 1 December 1996 (1996-12-01), GB, pages 2791 - 2794., XP002915911, ISSN: 0040-4039, DOI: 10.1016/0040-4039(96)00430-3 *
WIPF P., VENKATRAMAN S.: "AN IMPROVED PROTOCOL FOR AZOLE SYNTHESIS WITH PEG-SUPPORTED BURGESS REAGENT.", TETRAHEDRON LETTERS, PERGAMON, GB, vol. 37., no. 27., 1 December 1996 (1996-12-01), GB, pages 4659 - 4662., XP002915910, ISSN: 0040-4039, DOI: 10.1016/0040-4039(96)00918-5 *

Cited By (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7879576B2 (en) 2000-11-16 2011-02-01 Regents Of The University Of California Marine actinomycete taxon for drug and fermentation product discovery
US7144723B2 (en) 2000-11-16 2006-12-05 The Regents Of The University Of California Marine actinomycete taxon for drug and fermentation product discovery
US7928138B2 (en) 2000-11-16 2011-04-19 Regents Of The University Of California Marine actinomycete taxon for drug and fermentation product discovery
WO2002082091A3 (fr) * 2001-04-09 2004-03-04 Allan Christian Shaw Procede d'identification de proteines a partir de bacteries intracellulaires
US8222289B2 (en) 2002-06-24 2012-07-17 The Regents Of The University Of California Salinosporamides and methods for use thereof
US10314818B2 (en) 2002-06-24 2019-06-11 The Regents Of The University Of California Salinosporamides and methods of use thereof
US9078881B2 (en) 2002-06-24 2015-07-14 The Regents Of The University Of California Salinosporamides and methods of use thereof
US7635712B2 (en) 2002-06-24 2009-12-22 The Regents Of The University Of California Salinosporamides and methods for use thereof
US9713607B2 (en) 2002-06-24 2017-07-25 The Regents Of The University Of California Salinosporamides and methods of use thereof
US8637565B2 (en) 2002-06-24 2014-01-28 The Regents Of The University Of California Salinosporamides and methods for use thereof
US7179834B2 (en) 2002-06-24 2007-02-20 The Regents Of The University Of California Salinosporamides and methods for use thereof
US10912764B2 (en) 2002-06-24 2021-02-09 The Regents Of The University Of California Salinosporamides and methods of use thereof
US7176232B2 (en) 2002-06-24 2007-02-13 The Regents Of The University Of California Salinosporamides and methods for use thereof
WO2004012732A3 (fr) * 2002-07-31 2004-04-29 Charite Universitaetsmedizin Utilisation d'un inhibiteur du proteasome dans le traitement du dysfonctionnement endothelial et/ou dans le cadre d'une therapie a base de proteasome a faible dose
US8217072B2 (en) 2003-06-20 2012-07-10 The Regents Of The University Of California Salinosporamides and methods for use thereof
US8168803B2 (en) 2003-06-20 2012-05-01 Nereus Pharmaceuticals, Inc. Methods of using [3.2.0] heterocyclic compounds and analogs thereof
US7276530B2 (en) 2004-04-30 2007-10-02 Nereus Pharmaceuticals, Inc. [3.2.0] Heterocyclic compounds and methods of using the same
US7579371B2 (en) 2004-04-30 2009-08-25 Nereus Pharmaceuticals, Inc. Methods of using [3.2.0] heterocyclic compounds and analogs thereof
US7544814B2 (en) 2004-04-30 2009-06-09 Nereus Pharmaceuticals, Inc. [3.2.0] Heterocyclic compounds and methods of using the same
US8722724B2 (en) 2004-12-03 2014-05-13 Triphase Research And Development I Corp. Compositions and methods for treating neoplastic diseases
US10610517B2 (en) 2004-12-03 2020-04-07 Celgene International Ii Sàrl Compositions and methods for treating neoplastic diseases
US7842814B2 (en) 2006-04-06 2010-11-30 Nereus Pharmaceuticals, Inc. Total synthesis of salinosporamide A and analogs thereof
US8067616B2 (en) 2006-04-06 2011-11-29 Nereus Pharmaceuticals, Inc. Total synthesis of salinosporamide A and analogs thereof
US7824698B2 (en) 2007-02-02 2010-11-02 Nereus Pharmaceuticals, Inc. Lyophilized formulations of Salinosporamide A
US8394816B2 (en) 2007-12-07 2013-03-12 Irene Ghobrial Methods of using [3.2.0] heterocyclic compounds and analogs thereof in treating Waldenstrom's Macroglobulinemia
US8003802B2 (en) 2008-03-07 2011-08-23 Nereus Pharmaceuticals, Inc. Total synthesis of Salinosporamide A and analogs thereof
US8314251B2 (en) 2008-03-07 2012-11-20 Nereus Pharmaceuticals, Inc. Total synthesis of salinosporamide A and analogs thereof
US8389564B2 (en) 2008-05-12 2013-03-05 Venkat Rami Reddy Macherla Proteasome inhibitors
US8227503B2 (en) 2008-05-12 2012-07-24 Nereus Pharmaceuticals, Inc. Proteasome inhibitors
US7910616B2 (en) 2008-05-12 2011-03-22 Nereus Pharmaceuticals, Inc. Proteasome inhibitors

Also Published As

Publication number Publication date
AU8906298A (en) 1999-03-08
AU749857B2 (en) 2002-07-04
CA2301054A1 (fr) 1999-02-25
HUP0002724A2 (hu) 2001-02-28
KR20010022950A (ko) 2001-03-26
JP2001515064A (ja) 2001-09-18
CN1271342A (zh) 2000-10-25
HUP0002724A3 (en) 2001-04-28
NZ503169A (en) 2001-12-21
IL134538A (en) 2005-11-20
IL134538A0 (en) 2001-04-30
EP1021407A1 (fr) 2000-07-26
BR9811304A (pt) 2001-11-13
EP1021407A4 (fr) 2001-07-04

Similar Documents

Publication Publication Date Title
US6566553B2 (en) Synthesis of clasto-lactacystin β-lactone and analogs thereof
Seebach et al. EPC syntheses with C, C bond formation via acetals and enamines
DE69623785T2 (de) 4,5-diaryl oxazole derivate
WO1999009006A1 (fr) SYNTHESE DE CLASTO-LACTACYSTINE β-LACTONE ET DE SES ANALOGUES
US4929630A (en) Transglutaminase inhibitors
Yakura et al. Synthesis of an immunomodulator (+)-conagenin and its analogs
RU2423348C2 (ru) Синтез ингибиторов ренина с использованием реакции циклоприсоединения
AU2009263076B2 (en) New heterocyclic carboxamides for use as thrombin inhibitors
Groth et al. Asymmetric synthesis of D-erythro-sphingosine
US5770732A (en) Pyrrolinone-based peptidomimetics
US5668164A (en) Process for synthesis of chiral cis-and trans-3-amino-4 substituted pyrrolidine compounds
Cremonesi et al. Stereoselective synthesis of β-hydroxy-α-amino acids β-substituted with non-aromatic heterocycles
US6166227A (en) Preparation of (S)-2-amino-6, 6-dimethoxyhexanoic acid methyl ester via novel dioxolanes
US5514814A (en) Pyrrolinone-based compounds
MXPA00001551A (en) SYNTHESIS OF CLASTO-LACTACYSTIN&bgr;-LACTONE AND ANALOGS THEREOF
KR940000754B1 (ko) 리닌-저해성 올리고펩티드, 그의 제조방법 및 용도
EP1100799B1 (fr) Preparation de derives de la camptothecine et de la nothapodytine
WO1995008530A1 (fr) Procede de production d'un derive de 3-amino-2-hydroxy-1-propanol
Pilli et al. 27.10 Product Class 10: N-Acyliminium Salts
WO2003091210A1 (fr) Nouvelle synthese pour la preparation de porphobilinogene et compose en tant que sel stabilise et ses intermediaires
JPH0395168A (ja) ジペプチド化合物
JPH0250107B2 (fr)

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 134538

Country of ref document: IL

Ref document number: 98809526.2

Country of ref document: CN

AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH GM HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: PA/a/2000/001551

Country of ref document: MX

ENP Entry into the national phase

Ref document number: 2301054

Country of ref document: CA

Ref document number: 2301054

Country of ref document: CA

Kind code of ref document: A

Ref document number: 2000 509690

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 1020007001558

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 89062/98

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 503169

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 1998940885

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 1998940885

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1020007001558

Country of ref document: KR

WWG Wipo information: grant in national office

Ref document number: 89062/98

Country of ref document: AU

WWG Wipo information: grant in national office

Ref document number: 1020007001558

Country of ref document: KR

WWW Wipo information: withdrawn in national office

Ref document number: 1998940885

Country of ref document: EP