[go: up one dir, main page]

US20050283023A1 - Preparation of pregabalin and related compounds - Google Patents

Preparation of pregabalin and related compounds Download PDF

Info

Publication number
US20050283023A1
US20050283023A1 US11/157,585 US15758505A US2005283023A1 US 20050283023 A1 US20050283023 A1 US 20050283023A1 US 15758505 A US15758505 A US 15758505A US 2005283023 A1 US2005283023 A1 US 2005283023A1
Authority
US
United States
Prior art keywords
formula
compound
salt
alkyl
cyano
Prior art date
Legal status (The legal status 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 status listed.)
Abandoned
Application number
US11/157,585
Inventor
Shanghui Hu
Carlos Martinez
Junhua Tao
William Tully
Patrick Kelleher
Yves Dumond
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
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
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=35295367&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=US20050283023(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Individual filed Critical Individual
Priority to US11/157,585 priority Critical patent/US20050283023A1/en
Publication of US20050283023A1 publication Critical patent/US20050283023A1/en
Priority to US12/254,336 priority patent/US7838686B2/en
Priority to US12/951,429 priority patent/US8044227B2/en
Priority to US13/247,032 priority patent/US8134023B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C253/00—Preparation of carboxylic acid nitriles
    • C07C253/30—Preparation of carboxylic acid nitriles by reactions not involving the formation of cyano groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/12—Formation of amino and carboxyl groups
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/08—Antiepileptics; Anticonvulsants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/04—Formation of amino groups in compounds containing carboxyl groups
    • C07C227/06—Formation of amino groups in compounds containing carboxyl groups by addition or substitution reactions, without increasing the number of carbon atoms in the carbon skeleton of the acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/14—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
    • C07C227/18—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
    • C07C227/20—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters by hydrolysis of N-acylated amino-acids or derivatives thereof, e.g. hydrolysis of carbamates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/22—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from lactams, cyclic ketones or cyclic oximes, e.g. by reactions involving Beckmann rearrangement
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/24—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having more than one carboxyl group bound to the carbon skeleton, e.g. aspartic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/19—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms containing cyano groups and carboxyl groups, other than cyano groups, bound to the same saturated acyclic carbon skeleton
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/001—Amines; Imines
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/002—Nitriles (-CN)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/42—Hydroxy-carboxylic acids

Definitions

  • This invention relates to methods and materials for preparing enantiomerically-enriched ⁇ -amino acids via enzymatic kinetic resolution, and is particularly useful for preparing ⁇ -amino acids that exhibit binding affinity to the human ⁇ 2 ⁇ calcium channel subunit, including pregabalin and related compounds.
  • Pregabalin (S)-(+)-3-aminomethyl-5-methyl-hexanoic acid, is related to the endogenous inhibitory neurotransmitter ⁇ -aminobutyric acid (GABA), which is involved in the regulation of brain neuronal activity.
  • GABA ⁇ -aminobutyric acid
  • Pregabalin exhibits anti-seizure activity, as discussed in U.S. Pat. No. 5,563,175 to R. B. Silverman et al., and is thought to be useful for treating, among other conditions, pain, physiological conditions associated with psychomotor stimulants, inflammation, gastrointestinal damage, alcoholism, insomnia, and various psychiatric disorders, including mania and bipolar disorder. See, respectively, U.S. Pat. No. 6,242,488 to L.
  • Pregabalin has been prepared in various ways. Typically, a racemic mixture of 3-aminomethyl-5-methyl-hexanoic acid is synthesized and subsequently resolved into its R- and S-enantiomers. Such methods may employ an azide intermediate, a malonate intermediate, or Hofman synthesis. See, respectively, U.S. Pat. No. 5,563,175 to R. B. Silverman et al.; U.S. Pat. Nos. 6,046,353, 5,840,956, and 5,637,767 to T. M. Grote et al.; and U.S. Pat. Nos. 5,629,447 and 5,616,793 to B. K. Huckabee & D. M.
  • Pregabalin has also been synthesized directly using a chiral auxiliary, (4R,5S)-4-methyl-5-phenyl-2-oxazolidinone. See, e.g., U.S. Pat. Nos. 6,359,169, 6,028,214, 5,847,151, 5,710,304, 5,684,189, 5,608,090, and 5,599,973, all to R. B. Silverman et al, which are herein incorporated by reference in their entirety and for all purposes.
  • a recently published U.S. patent application discusses a method of making pregabalin via asymmetric hydrogenation of a cyano-substituted olefin to produce a chiral cyano precursor of (S)-3-aminomethyl-5-methylhexanoic acid. See commonly assigned U.S. Patent Application No. 2003/0212290 A1 to Burk et al., published Nov. 13, 2003, which is herein incorporated by reference in its entirety for all purposes.
  • the cyano precursor is subsequently reduced to give pregabalin.
  • the asymmetric hydrogenation employs a chiral catalyst that is comprised of a transition metal bound to a bisphosphine ligand, such as (R,R)-Me-DUPHOS.
  • the method results in substantial enrichment of pregabalin over (R)-3-(aminomethyl)-5-methylhexanoic acid.
  • the present invention provides materials and methods for preparing enantiomerically enriched ⁇ -amino acids (Formula 1) such as pregabalin (Formula 9).
  • the method of the present invention involves a kinetic resolution of a racemic cyano diester intermediate (Formula 4 or Formula 12) using an enzyme that is adapted to enantioselectively hydrolyze an ester moiety of the intermediate.
  • the resulting dicarboxylic acid monoester (Formula 3 or Formula 11), which is substantially enantiopure, undergoes further reaction to yield the desired enantiomerically-enriched ⁇ -amino acids (Formula 1 or Formula 9).
  • the unreacted enantiomer (Formula 5 or Formula 13) from the kinetic resolution can be reused in the enzymatic resolution following racemization, thereby improving overall yield.
  • the claimed method offers significant advantages over existing processes for preparing enantiomerically-enriched ⁇ -amino acids (Formula 1 and Formula 9).
  • the optically-active ⁇ -amino acids can be prepared without using chiral auxiliaries or proprietary hydrogenation catalysts, which should lead to lower unit costs. Since enzymatic processes can be carried out at room temperature and at atmospheric pressure, the claimed methods should help minimize scheduling conflicts arising from the use of specialized equipment capable of handling high pressures and low temperatures.
  • the present invention can be used to prepare pregabalin starting from a racemic cyano-substituted diester (Formula 12) in good yield (26% to 31%) after a single batch recycle of the unreacted enantiomer (Formula 13). This translates into about a 50% savings in cost of goods over the malonate method described above.
  • One aspect of the present invention provides a method of making a compound of or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, in which
  • Another aspect of the present invention provides a method of making a compound of Formula 1, above, the method comprising:
  • the compound of Formula 6, above may be prepared by hydrolyzing a compound of Formula 3, or a salt thereof, wherein R 1 and R 2 in Formula 3 are as defined above in Formula 1, and R 3 is C 1-12 alkyl, C 3-12 cycloalkyl, or aryl-C 1-6 alkyl.
  • An additional aspect of the present invention provides method of making a compound of Formula 1, above, the method comprising:
  • the compound of Formula 8 may be prepared by decarboxylating a compound of Formula 3, above, or a salt thereof, or by hydrolyzing and decarboxylating the compound of Formula 3 or a salt thereof, to yield the compound of Formula 8 or a salt thereof.
  • a further aspect of the present invention provides a method of making the compound of Formula 3, above, or a salt thereof, the method comprising:
  • Any number of enzymes may be used to enantioselectively hydrolyze the compound of Formula 4 to the compound of Formula 3 or a salt thereof.
  • Useful enzymes include lipases, such as those derived from Thermomyces lanuginosus.
  • Another aspect of the present invention provides compounds represented by Formula 2, above, including complexes, salts, solvates or hydrates thereof, provided that when one of the substituents represented by R 1 or R 2 in Formula 2 is hydrogen, the other substituent is not C 1-3 alkyl or C 5 alkyl.
  • An additional aspect of the present invention provides compounds of Formula 27, including complexes, salts, solvates or hydrates thereof, wherein
  • Compounds of Formula 27 include those represented by Formula 3, Formula 4, Formula 5, Formula 6, and Formula 7, above, including their complexes, salts, solvates or hydrates.
  • Useful compounds of Formula 2-7 and 27 include those in which R 1 is a hydrogen atom and R is isobutyl.
  • a further aspect of the present invention provides a method of making a compound of Formula 9, or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
  • the compound of Formula 10 or a salt thereof may be prepared by reducing a cyano moiety of a compound of Formula 11, or a salt thereof, wherein R 3 is as defined above in Formula 3.
  • Another aspect of the present invention provides a method of making a compound of Formula 9, above, or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
  • the compound of Formula 14, above may be prepared by hydrolyzing a compound of Formula 11, or salt thereof, wherein R 3 in Formula 11 is as defined above in Formula 3.
  • An additional aspect of the present invention provides a method of making a compound of Formula 9, above, or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
  • the compound of Formula 16 may be prepared by decarboxylating (e.g., by heating) the compound of Formula 11, above, or a salt thereof, or by hydrolyzing and decarboxylating the compound of Formula 11 or a salt thereof.
  • a further aspect of the present invention provides a method of making the compound of Formula 11, above, or a salt thereof, the method comprising:
  • the corresponding salts of the compound of Formula 11 include those selected from alkali metal salts, such as potassium salt; primary amine salts, such as a t-butyl amine salt; and secondary amine salts.
  • useful enzymes include lipases, such as those derived from Thermomyces lanuginosus.
  • Another aspect of the present invention provides a compound selected from:
  • the present invention includes all complexes and salts, whether pharmaceutically acceptable or not, solvates, hydrates, and polymorphic forms of the disclosed compounds.
  • Certain compounds may contain an alkenyl or cyclic group, so that cisltrans (or Z/E) stereoisomers are possible, or may contain a keto or oxime group, so that tautomerism may occur.
  • the present invention generally includes all Z/E isomers and tautomeric forms, whether they are pure, substantially pure, or mixtures.
  • FIG. 1 depicts a scheme for preparing enantiomerically-enriched ⁇ -amino acids (Formula 1).
  • FIG. 2 depicts a scheme for preparing cyano-substituted diesters (Formula 4).
  • ⁇ a dash (“ ⁇ ”) to indicate a bond between atoms or a point of attachment to a named or unnamed atom or group of atoms.
  • Certain formulae may also include one or more asterisks (“*”) to indicate stereogenic (asymmetric or chiral) centers, although the absence of an asterisk does not indicate that the compound lacks a stereocenter.
  • Such formulae may refer to the racemate or to individual enantiomers or to individual diastereomers, which may or may not be pure or substantially pure.
  • “Substituted” groups are those in which one or more hydrogen atoms have been replaced with one or more non-hydrogen groups, provided that valence requirements are met and that a chemically stable compound results from the substitution.
  • Alkyl refers to straight chain and branched saturated hydrocarbon groups, generally having a specified number of carbon atoms (i.e., C 1-6 alkyl refers to an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms and C 1-12 alkyl refers to an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms).
  • alkyl groups include, without limitation, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, pent-1-yl, pent-2-yl, pent-3-yl, 3-methylbut-1-yl, 3-methylbut-2-yl, 2-methylbut-2-yl, 2,2,2-trimethyleth-1-yl, n-hexyl, and the like.
  • Alkenyl refers to straight chain and branched hydrocarbon groups having one or more unsaturated carbon-carbon bonds, and generally having a specified number of carbon atoms.
  • alkenyl groups include, without limitation, ethenyl, 1-propen-1-yl, 1-propen-2-yl, 2-propen-1-yl, 1-buten-1-yl, 1-buten-2-yl, 3-buten-1-yl, 3-buten-2-yl, 2-buten-1-yl, 2-buten-2-yl, 2-methyl-1-propen-1-yl, 2-methyl-2-propen-1-yl, 1,3-butadien-1-yl, 1,3-butadien-2-yl, and the like.
  • Alkynyl refers to straight chain or branched hydrocarbon groups having one or more triple carbon-carbon bonds, and generally having a specified number of carbon atoms. Examples of alkynyl groups include, without limitation, ethynyl, 1-propyn-1-yl, 2-propyn-1-yl, 1-butyn-1-yl, 3-butyn-1-yl, 3-butyn-2-yl, 2-butyn-1-yl, and the like.
  • alkanoyl and alkanoylamino refer, respectively, to alkyl-C(O)— and alkyl-C(O)—NH—, where alkyl is defined above, and generally includes a specified number of carbon atoms, including the carbonyl carbon.
  • alkanoyl groups include, without limitation, formyl, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, and the like.
  • alkenoyl and alkynoyl refer, respectively, to alkenyl-C(O)— and alkynyl-C(O)—, where alkenyl and alkynyl are defined above. References to alkenoyl and alkynoyl generally include a specified number of carbon atoms, excluding the carbonyl carbon. Examples of alkenoyl groups include, without limitation, propenoyl, 2-methylpropenoyl, 2-butenoyl, 3-butenoyl, 2-methyl-2-butenoyl, 2-methyl-3-butenoyl, 3-methyl-3-butenoyl, 2-pentenoyl, 3-pentenoyl, 4-pentenoyl, and the like.
  • alkynoyl groups include, without limitation, propynoyl, 2-butynoyl, 3-butynoyl, 2-pentynoyl, 3-pentynoyl, 4-pentynoyl, and the like.
  • Alkoxy refers, respectively, to alkyl-O—, alkenyl-O, and alkynyl-O; to alkyl-O—C(O)—, alkenyl-O—C(O)—, alkynyl-O—C(O)—; and to alkyl-O—C(O)—NH—, alkenyl-O—C(O)—NH—, and alkynyl-O—C(O)—NH—, where alkyl, alkenyl, and alkynyl are defined above.
  • alkoxy groups include, without limitation, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, n-pentoxy, s-pentoxy, and the like.
  • alkoxycarbonyl groups include, without limitation, methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, i-propoxycarbonyl, n-butoxycarbonyl, s-butoxycarbonyl, t-butoxycarbonyl, n-pentoxycarbonyl, s-pentoxycarbonyl, and the like.
  • Alkylamino refers, respectively, to alkyl-NH—, alkyl-NH—C(O)—, alkyl 2 -N—C(O)—, alkyl-S(O 2 )—, HS(O 2 )—NH-alkyl-, and alkyl-S(O)—NH—C(O)— where alkyl is defined above.
  • Aminoalkyl and cyanoalkyl refer, respectively, to NH 2 -alkyl and N ⁇ C-alkyl, where alkyl is defined above.
  • Halo “Halo,” “halogen” and “halogeno” may be used interchangeably, and refer to fluoro, chloro, bromo, and iodo.
  • Haloalkyl refers, respectively, to alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, alkoxy, and alkoxycarbonyl groups substituted with one or more halogen atoms, where alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, alkoxy, and alkoxycarbonyl are defined above.
  • haloalkyl groups include, without limitation, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, and the like.
  • Examples of hydroxyalkyl and oxoalkyl groups include, without limitation, hydroxymethyl, hydroxyethyl, 3-hydroxypropyl, oxomethyl, oxoethyl, 3-oxopropyl, and the like.
  • Cycloalkyl refers to saturated monocyclic and bicyclic hydrocarbon rings, generally having a specified number of carbon atoms that comprise the ring (i.e., C 3-7 cycloalkyl refers to a cycloalkyl group having 3, 4, 5, 6 or 7 carbon atoms as ring members).
  • the cycloalkyl may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements.
  • the cycloalkyl groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements.
  • Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, alkoxy, alkoxycarbonyl, alkanoyl, and halo, as defined above, and hydroxy, mercapto, nitro, and amino.
  • Examples of monocyclic cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
  • Examples of bicyclic cycloalkyl groups include, without limitation, bicyclo[1.1.0]butyl, bicyclo[1.1.1]pentyl, bicyclo[2.1.0]pentyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.0]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.0]heptyl, bicyclo[3.1.1]heptyl, bicyclo[4.1.0]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.1.1]octyl, bicyclo[3.3.0]octyl, bicyclo[4.2.0]octyl, bicyclo[3.3.1]nonyl,
  • Cycloalkenyl refers monocyclic and bicyclic hydrocarbon rings having one or more unsaturated carbon-carbon bonds and generally having a specified number of carbon atoms that comprise the ring (i.e., C 3-7 cycloalkenyl refers to a cycloalkenyl group having 3, 4, 5, 6 or 7 carbon atoms as ring members).
  • the cycloalkenyl may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements.
  • the cycloalkenyl groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements.
  • Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, alkoxy, alkoxycarbonyl, alkanoyl, and halo, as defined above, and hydroxy, mercapto, nitro, and amino.
  • “Cycloalkanoyl” and “cycloalkenoyl” refer to cycloalkyl-C(O)— and cycloalkenyl-C(O)—, respectively, where cycloalkyl and cycloalkenyl are defined above. References to cycloalkanoyl and cycloalkenoyl generally include a specified number of carbon atoms, excluding the carbonyl carbon.
  • cycloalkanoyl groups include, without limitation, cyclopropanoyl, cyclobutanoyl, cyclopentanoyl, cyclohexanoyl, cycloheptanoyl, 1-cyclobutenoyl, 2-cyclobutenoyl, 1-cyclopentenoyl, 2-cyclopentenoyl, 3-cyclopentenoyl, 1-cyclohexenoyl, 2-cyclohexenoyl, 3-cyclohexenoyl, and the like.
  • Cycloalkoxy and “cycloalkoxycarbonyl” refer, respectively, to cycloalkyl-O— and cycloalkenyl-O and to cycloalkyl-O—C(O)— and cycloalkenyl-O—C(O)—, where cycloalkyl and cycloalkenyl are defined above.
  • References to cycloalkoxy and cycloalkoxycarbonyl generally include a specified number of carbon atoms, excluding the carbonyl carbon.
  • cycloalkoxy groups include, without limitation, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexoxy, 1-cyclobutenoxy, 2-cyclobutenoxy, 1-cyclopentenoxy, 2-cyclopentenoxy, 3-cyclopentenoxy, 1-cyclohexenoxy, 2-cyclohexenoxy, 3-cyclohexenoxy, and the like.
  • cycloalkoxycarbonyl groups include, without limitation, cyclopropoxycarbonyl, cyclobutoxycarbonyl, cyclopentoxycarbonyl, cyclohexoxycarbonyl, 1-cyclobutenoxycarbonyl, 2-cyclobutenoxycarbonyl, 1-cyclopentenoxycarbonyl, 2-cyclopentenoxycarbonyl, 3-cyclopentenoxycarbonyl, 1-cyclohexenoxycarbonyl, 2-cyclohexenoxycarbonyl, 3-cyclohexenoxycarbonyl, and the like.
  • Aryl and “arylene” refer to monovalent and divalent aromatic groups, respectively, including 5- and 6-membered monocyclic aromatic groups that contain 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.
  • monocyclic aryl groups include, without limitation, phenyl, pyrrolyl, furanyl, thiopheneyl, thiazolyl, isothiazolyl, imidazolyl, triazolyl, tetrazolyl, pyrazolyl, oxazolyl, isooxazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, and the like.
  • Aryl and arylene groups also include bicyclic groups, tricyclic groups, etc., including fused 5- and 6-membered rings described above.
  • multicyclic aryl groups include, without limitation, naphthyl, biphenyl, anthracenyl, pyrenyl, carbazolyl, benzoxazolyl, benzodioxazolyl, benzothiazolyl, benzoimidazolyl, benzothiopheneyl, quinolinyl, isoquinolinyl, indolyl, benzofuranyl, purinyl, indolizinyl, and the like.
  • aryl and arylene groups may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements.
  • aryl and arylene groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements.
  • Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, alkanoyl, cycloalkanoyl, cycloalkenoyl, alkoxycarbonyl, cycloalkoxycarbonyl, and halo, as defined above, and hydroxy, mercapto, nitro, amino, and alkylamino.
  • Heterocycle and “heterocyclyl” refer to saturated, partially unsaturated, or unsaturated monocyclic or bicyclic rings having from 5 to 7 or from 7 to 11 ring members, respectively. These groups have ring members made up of carbon atoms and from 1 to 4 heteroatoms that are independently nitrogen, oxygen or sulfur, and may include any bicyclic group in which any of the above-defined monocyclic heterocycles are fused to a benzene ring. The nitrogen and sulfur heteroatoms may optionally be oxidized. The heterocyclic ring may be attached to a parent group or to a substrate at any heteroatom or carbon atom unless such attachment would violate valence requirements.
  • any of the carbon or nitrogen ring members may include a non-hydrogen substituent unless such substitution would violate valence requirements.
  • Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, alkanoyl, cycloalkanoyl, cycloalkenoyl, alkoxycarbonyl, cycloalkoxycarbonyl, and halo, as defined above, and hydroxy, mercapto, nitro, amino, and alkylamino.
  • heterocycles include, without limitation, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indoli
  • Heteroaryl and heteroarylene refer, respectively, to monovalent and divalent heterocycles or heterocyclyl groups, as defined above, which are aromatic. Heteroaryl and heteroarylene groups represent a subset of aryl and arylene groups, respectively.
  • Arylalkyl and “heteroarylalkyl” refer, respectively, to aryl-alkyl and heteroaryl-alkyl, where aryl, heteroaryl, and alkyl are defined above. Examples include, without limitation, benzyl, fluorenylmethyl, imidazol-2-yl-methyl, and the like.
  • Arylalkanoyl refers, respectively, to aryl-alkanoyl, heteroaryl-alkanoyl, aryl-alkenoyl, heteroaryl-alkenoyl, aryl-alkynoyl, and heteroaryl-alkynoyl, where aryl, heteroaryl, alkanoyl, alkenoyl, and alkynoyl are defined above.
  • Examples include, without limitation, benzoyl, benzylcarbonyl, fluorenoyl, fluorenylmethylcarbonyl, imidazol-2-oyl, imidazol-2-yl-methylcarbonyl, phenylethenecarbonyl, 1-phenylethenecarbonyl, 1-phenyl-propenecarbonyl, 2-phenyl-propenecarbonyl, 3-phenyl-propenecarbonyl, imidazol-2-yl-ethenecarbonyl, 1-(imidazol-2-yl)-ethenecarbonyl, 1-(imidazol-2-yl)-propenecarbonyl, 2-(imidazol-2-yl)-propenecarbonyl, 3-(imidazol-2-yl)-propenecarbonyl, phenylethynecarbonyl, phenylpropynecarbonyl, (imidazol-2-yl)-ethynecarbonyl,
  • Arylalkoxy and “heteroarylalkoxy” refer, respectively, to aryl-alkoxy and heteroaryl-alkoxy, where aryl, heteroaryl, and alkoxy are defined above. Examples include, without limitation, benzyloxy, fluorenylmethyloxy, imidazol-2-yl-methyloxy, and the like.
  • Aryloxy and “heteroaryloxy” refer, respectively, to aryl-O— and heteroaryl-O—, where aryl and heteroaryl are defined above. Examples include, without limitation, phenoxy, imidazol-2-yloxy, and the like.
  • Aryloxycarbonyl,” “heteroaryloxycarbonyl,” “arylalkoxycarbonyl,” and “heteroarylalkoxycarbonyl” refer, respectively, to aryloxy-C(O)—, heteroaryloxy-C(O)—, arylalkoxy-C(O)—, and heteroarylalkoxy-C(O)—, where aryloxy, heteroaryloxy, arylalkoxy, and heteroarylalkoxy are defined above. Examples include, without limitation, phenoxycarbonyl, imidazol-2-yloxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, imidazol-2-yl-methyloxycarbonyl, and the like.
  • Leaving group refers to any group that leaves a molecule during a fragmentation process, including substitution reactions, elimination reactions, and addition-elimination reactions. Leaving groups may be nucleofugal, in which the group leaves with a pair of electrons that formerly served as the bond between the leaving group and the molecule, or may be electrofugal, in which the group leaves without the pair of electrons. The ability of a nucleofugal leaving group to leave depends on its base strength, with the strongest bases being the poorest leaving groups.
  • Common nucleofugal leaving groups include nitrogen (e.g., from diazonium salts); sulfonates, including alkylsulfonates (e.g., mesylate), fluoroalkylsulfonates (e.g., triflate, hexaflate, nonaflate, and tresylate), and arylsulfonates (e.g., tosylate, brosylate, closylate, and nosylate). Others include carbonates, halide ions, carboxylate anions, phenolate ions, and alkoxides. Some stronger bases, such as NH 2 ⁇ and OH ⁇ can be made better leaving groups by treatment with an acid. Common electrofugal leaving groups include the proton, CO 2 , and metals.
  • Enantiomeric excess or “ee” is a measure, for a given sample, of the excess of one enantiomer over a racemic sample of a chiral compound and is expressed as a percentage. Enantiomeric excess is defined as 100 ⁇ (er ⁇ 1)/(er+1), where “er” is the ratio of the more abundant enantiomer to the less abundant enantiomer.
  • Diastereomeric excess or “de” is a measure, for a given sample, of the excess of one diastereomer over a sample having equal amounts of diastereomers and is expressed as a percentage. Diastereomeric excess is defined as 100 ⁇ (dr ⁇ 1)/(dr+1), where “dr” is the ratio of a more abundant diastereomer to a less abundant diastereomer.
  • Stepselective refer to a given process (e.g., ester hydrolysis, hydrogenation, hydroformylation, ⁇ -allyl palladium coupling, hydrosilation, hydrocyanation, olefin metathesis, hydroacylation, allylamine isomerization, etc.) that yields more of one stereoisomer, enantiomer, or diastereoisomer than of another, respectively.
  • a given process e.g., ester hydrolysis, hydrogenation, hydroformylation, ⁇ -allyl palladium coupling, hydrosilation, hydrocyanation, olefin metathesis, hydroacylation, allylamine isomerization, etc.
  • “High level of stereoselectivity,” “high level of enantioselectivity,” “high level of diastereoselectivity,” and variants thereof, refer to a given process that yields products having an excess of one stereoisomer, enantiomer, or diastereoisomer, which comprises at least about 90% of the products.
  • a high level of enantioselectivity or diastereoselectivity would correspond to an ee or de of at least about 80%.
  • Stepoisomerically enriched refers, respectively, to a sample of a compound that has more of one stereoisomer, enantiomer or diastereomer than another.
  • the degree of enrichment may be measured by % of total product, or for a pair of enantiomers or diastereomers, by ee or de.
  • substantially pure stereoisomer refers, respectively, to a sample containing a stereoisomer, enantiomer, or diastereomer, which comprises at least about 95% of the sample.
  • a substantially pure enantiomer or diastereomer would correspond to samples having an ee or de of about 90% or greater.
  • a “pure stereoisomer,” “pure enantiomer,” “pure diastereomer,” and variants thereof, refer, respectively, to a sample containing a stereoisomer, enantiomer, or diastereomer, which comprises at least about 99.5% of the sample.
  • a pure enantiomer or pure diastereomer would correspond to samples having an ee or de of about 99% or greater.
  • Opte enantiomer refers to a molecule that is a non-superimposable mirror image of a reference molecule, which may be obtained by inverting all of the stereogenic centers of the reference molecule. For example, if the reference molecule has S absolute stereochemical configuration, then the opposite enantiomer has R absolute stereochemical configuration. Likewise, if the reference molecule has S,S absolute stereochemical configuration, then the opposite enantiomer has R,R stereochemical configuration, and so on.
  • Stepoisomers of a specified compound refer to the opposite enantiomer of the compound and to any diastereoisomers or geometric isomers (ZiE) of the compound.
  • ZiE geometric isomers
  • the specified compound has S,R,Z stereochemical configuration
  • its stereoisomers would include its opposite enantiomer having R,S,Z configuration, its diastereomers having S,S,Z configuration and R,R,Z configuration
  • its geometric isomers having S,R,E configuration, R,S,E configuration, S,S,E configuration, and R,R,E configuration.
  • “Lipase Unit” or “LU” refers to the amount of enzyme (in g) that liberates 1 ⁇ mol of titratable butyric acid/min when contacted with tributyrin and an emulsifier (gum arabic) at 30° C. and pH 7.
  • Solvate refers to a molecular complex comprising a disclosed or claimed compound and a stoichiometric or non-stoichiometric amount of one or more solvent molecules (e.g., EtOH).
  • solvent molecules e.g., EtOH
  • “Hydrate” refers to a solvate comprising a disclosed or claimed compound and a stoichiometric or non-stoichiometric amount of water.
  • “Pharmaceutically acceptable complexes, salts, solvates, or hydrates” refers to complexes, acid or base addition salts, solvates or hydrates of claimed and disclosed compounds, which are within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use.
  • Pre-catalyst or “catalyst precursor” refers to a compound or set of compounds that are converted into a catalyst prior to use.
  • Treating refers to reversing, alleviating, inhibiting the progress of, or preventing a disorder or condition to which such term applies, or to preventing one or more symptoms of such disorder or condition.
  • Treatment refers to the act of “treating,” as defined immediately above.
  • certain compounds can be prepared using protecting groups, which prevent undesirable chemical reaction at otherwise reactive sites.
  • Protecting groups may also be used to enhance solubility or otherwise modify physical properties of a compound.
  • protecting group strategies a description of materials and methods for installing and removing protecting groups, and a compilation of useful protecting groups for common functional groups, including amines, carboxylic acids, alcohols, ketones, aldehydes, and the like, see T. W. Greene and P. G. Wuts, Protecting Groups in Organic Chemistry (1999) and P. Kocienski, Protective Groups (2000), which are herein incorporated by reference in their entirety for all purposes.
  • the chemical transformations described throughout the specification may be carried out using substantially stoichiometric amounts of reactants, though certain reactions may benefit from using an excess of one or more of the reactants. Additionally, many of the reactions disclosed throughout the specification, including the enantioselective hydrolysis of the racemic diester (Formula 4) described in detail below, may be carried out at about RT, but particular reactions may require the use of higher or lower temperatures, depending on reaction kinetics, yields, and the like. Furthermore, many of the chemical transformations may employ one or more compatible solvents, which may influence the reaction rate and yield.
  • the one or more solvents may be polar protic solvents, polar aprotic solvents, non-polar solvents, or some combination. Any references in the disclosure to a concentration range, a temperature range, a pH range, a catalyst loading range, and so on, whether expressly using the word “range” or not, include the indicated endpoints.
  • the present invention provides materials and methods for preparing optically active ⁇ -amino acids (Formula 1) including pharmaceutically acceptable salts, esters, amides, or prodrugs thereof.
  • the compounds of Formula 1 include substituents R 1 and R 2 , which are defined above.
  • Useful compounds of Formula 1 thus include those in which R 1 is a hydrogen atom and R 2 is C 1-12 alkyl, C 3-12 cycloalkyl, or substituted C 3-12 cycloalkyl, or those in which R 2 is a hydrogen atom and R 1 is C 1-12 alkyl, C 3-1 2 cycloalkyl, or substituted C 3-12 cycloalkyl.
  • Particularly useful compounds of Formula 1 include those in which R 1 is a hydrogen atom and R 2 is C 1-6 alkyl or C 3-7 cycloalkyl, or those in which R 2 is a hydrogen atom and R 1 is C 1-6 alkyl or C 3-7 cycloalkyl.
  • Particular useful compounds of Formula 1 include those in which R 1 is a hydrogen atom and R 2 is C 1-4 alkyl, such as pregabalin (Formula 9).
  • FIG. 1 shows a process for preparing optically active ⁇ -amino acids (Formula 1).
  • the process includes the step of contacting or combining a reaction mixture, which is comprised of a cyano-substituted diester (Formula 4) and water, with an enzyme to yield a product mixture that includes an optically active dicarboxylic acid monoester (Formula 3) and an optically-active diester (Formula 5).
  • the cyano-substituted diester (Formula 4) has a stereogenic center, which is denoted by an asterisk (“*”), and as described below, may be prepared in accordance with a reaction scheme shown in FIG. 2 .
  • the cyano-substituted diester Prior to contacting the enzyme, typically comprises a racemic (equimolar) mixture of the diester of Formula 5 and its opposite enantiomer.
  • Substituents R 1 , R 2 , and R 3 in Formula 3, Formula 4, and Formula 5, and substituent R 4 in Formula 4 and Formula 5 are as defined above in connection with Formula 1.
  • substituent identifier R 1 , R 2 , R 3 , etc.
  • the enzyme may be any protein that, while having little or no effect on the compound of Formula 5, will catalyze the hydrolysis of its opposite enantiomer to yield the dicarboxylic acid monoester (Formula 3).
  • Useful enzymes for enantioselectively hydrolyzing the compound of Formula 4 to Formula 3 may thus include hydrolases, including lipases, certain proteases, and other enantioselective esterases.
  • Such enzymes may be obtained from a variety of natural sources, including animal organs and microorganisms. See, e.g., Table 2 for a non-limiting list of commercially available hydrolases.
  • Protease BioCatalytics101 Pseudomonas sp. Lipase BioCatalytics103 Fungal Lipase BioCatalytics105 Microbial, lyophilized Lipase BioCatalytics108 CAL-B, lyophilized BioCatalytics110 Candida sp., lyophilized BioCatalytics111 CAL-A, lyophilized BioCatalytics112 Thermomyces sp.
  • useful enzymes for the enantioselective conversion of the cyano-substituted diester (Formula 4 and Formula 12) to the desired optically active dicarboxylic acid monoester (Formula 3 and Formula 11) include lipases.
  • Particularly useful lipases include enzymes derived from the microorganism Thermomyces lanuginosus , such as those available from Novo-Nordisk A/S under the trade name LIPOLASE® (CAS no. 9001-62-1).
  • LIPOLASE® enzymes are obtained by submerged fermentation of an Aspergillus oryzae microorganism genetically modified with DNA from Thermomyces lanuginosus DSM 4109 that encodes the amino acid sequence of the lipase.
  • LIPOLASE® 100L and LIPOLASE® 100T are available as a liquid solution and a granular solid, respectively, each having a nominal activity of 100 kLU/g.
  • Other forms of LIPOLASE® include LIPOLASE® 50L, which has half the activity of LIPOLASE® 100L, and LIPOZYME® 100L, which has the same activity of LIPOLASE® 100L, but is food grade.
  • Suitable enzymes For example, large numbers of commercially available enzymes may be screened using high throughput screening techniques described in the Example section below. Other enzymes (or microbial sources of enzymes) may be screened using enrichment isolation techniques. Such techniques typically involve the use of carbon-limited or nitrogen-limited media supplemented with an enrichment substrate, which may be the racemic substrate (Formula 4) or a structurally similar compound. Potentially useful microorganisms are selected for further investigation based on their ability to grow in media containing the enrichment substrate.
  • an enrichment substrate which may be the racemic substrate (Formula 4) or a structurally similar compound.
  • microorganisms are subsequently evaluated for their ability to enantioselectively catalyze ester hydrolysis by contacting suspensions of the microbial cells with the racemic substrate and testing for the presence of the desired optically-active dicarboxylic acid monoester (Formula 3) using analytical methods such as chiral HPLC, gas-liquid chromatography, LC/MS, and the like.
  • enzyme engineering may be employed to improve the properties of the enzyme it produces.
  • enzyme engineering may be used to increase the yield and the enantioselectivity of the ester hydrolysis, to broaden the temperature and pH operating ranges of the enzyme, and to improve the enzyme's tolerance to organic solvents.
  • Useful enzyme engineering techniques include rational design methods, such as site-directed mutagenesis, and in vitro-directed evolution techniques that utilize successive rounds of random mutagenesis, gene expression, and high throughput screening to optimize desired properties. See, e.g., K. M. Koeller & C.-H. Wong, “Enzymes for chemical synthesis,” Nature 409:232-240 (11 Jan. 2001), and references cited therein, the complete disclosures of which are herein incorporated by reference.
  • the enzyme may be in the form of whole microbial cells, permeabilized microbial cells, extracts of microbial cells, partially purified enzymes, purified enzymes, and the like.
  • the enzyme may comprise a dispersion of particles having an average particle size, based on volume, of less than about 0.1 mm (fine dispersion) or of about 0.1 mm or greater (coarse dispersion).
  • coarse enzyme particles may be used repeatedly in batch processes, or in semi-continuous or continuous processes, and may usually be separated (e.g., by filtration) from other components of the bioconversion more easily than fine dispersions of enzymes.
  • Useful coarse enzyme dispersions include cross-linked enzyme crystals (CLECs) and cross-linked enzyme aggregates (CLEAs), which are comprised primarily of the enzyme. Other coarse dispersions may include enzymes immobilized on or within an insoluble support.
  • Useful solid supports include polymer matrices comprised of calcium alginate, polyacrylamide, EUPERGIT®, and other polymeric materials, as well as inorganic matrices, such as CELITE®.
  • CLECs and other enzyme immobilization techniques see U.S. Pat. No. 5,618,710 to M. A. Navia & N. L. St. Clair.
  • CLEAs including their preparation and use, see U.S. Patent Application No.
  • the reaction mixture may comprise a single phase or may comprise multiple phases (e.g., a two- or a three-phase system).
  • the enantioselective hydrolysis shown in FIG. 1 may take place in a single aqueous phase, which contains the enzyme, the initially racemic substrate (Formula 4), the undesired optically-active diester (Formula 5), and the desired optically-active dicarboxylic acid monoester (Formula 3).
  • the reaction mixture may comprise a multi-phase system that includes an aqueous phase in contact with a solid phase (e.g., enzyme or product), an aqueous phase in contact with an organic phase, or an aqueous phase in contact with an organic phase and a solid phase.
  • the enantioselective hydrolysis may be carried out in a two-phase system comprised of a solid phase, which contains the enzyme, and an aqueous phase, which contains the initially racemic substrate, the undesired optically-active diester, and the desired optically-active dicarboxylic acid monoester.
  • the enantioselective hydrolysis may be carried out in a three-phase system comprised of a solid phase, which contains the enzyme, an organic phase that initially contains the racemic substrate (Formula 4), and an aqueous phase that initially contains a small fraction of the racemic substrate.
  • a solid phase which contains the enzyme
  • an organic phase that initially contains the racemic substrate (Formula 4)
  • an aqueous phase that initially contains a small fraction of the racemic substrate.
  • the desired optically-active dicarboxylic acid monoester has a lower pKa than the unreacted optically-active diester (Formula 5) and therefore exhibits greater aqueous solubility
  • the organic phase becomes enriched in the unreacted diester while the aqueous phase becomes enriched in the desired dicarboxylic acid monoester as the reaction proceeds.
  • the amounts of the racemic substrate (Formula 4) and the biocatalyst used in the enantioselective hydrolysis will depend on, among other things, the properties of the particular cyano-substituted diester and enzyme. Generally, however, the reaction may employ a substrate having an initial concentration of about 0.1 M to about 3.0 M, and in many cases, having an initial concentration of about 1.5 M to about 3.0 M. Additionally, the reaction may generally employ an enzyme loading of about 1% to about 10%, and in many cases, may employ an enzyme loading of about 3% to about 4% (v/v).
  • the enantioselective hydrolysis may be carried out over wide ranges of temperature and pH.
  • the reaction may be carried out at a temperature of about 10° C. to a temperature of about 50° C., but is typically carried out at about RT.
  • Such temperatures generally permit substantially full conversion (e.g., about 42% to about 50%) of the racemate (Formula 4) in a reasonable amount of time (about 2 h to about 24 h) without deactivating the enzyme.
  • the enantioselective hydrolysis may be carried out at a pH of about 5 to a pH of about 10, more typically at a pH of about 6 to a pH of about 9, and often at a pH of about 6.5 to a pH of about 7.5.
  • the hydrolysis reaction may be run with internal pH control (i.e., in the presence of a suitable buffer) or may be run with external pH control through the addition of a base.
  • suitable buffers include potassium phosphate, sodium phosphate, sodium acetate, ammonium acetate, calcium acetate, BES, BICINE, HEPES, MES, MOPS, PIPES, TAPS, TES, TRICINE, Tris, TRIZMA®, or other buffers having a pKa of about 6 to a pKa of about 9.
  • the buffer concentration generally ranges from about 5 mM to about 1 mM, and typically ranges from about 50 mM to about 200 mM.
  • Suitable bases include aqueous solutions comprised of KOH, NaOH, NH 4 OH, etc., having concentrations ranging from about 0.5 M to about 15 M, or more typically, ranging from about 5 M to about 10 M.
  • Other inorganic additives such as calcium acetate may also be used.
  • the desired optically active dicarboxylic acid monoester (Formula 3) is isolated from the product mixture using standard techniques.
  • the product mixture may be extracted one or more times with a nonpolar organic solvent, such as hexane or heptane, which separates the desired dicarboxylic monoester (Formula 2) and the unreacted diester (Formula 5) in aqueous and organic phases, respectively.
  • the monoester and diester may be separated batch-wise following reaction, or may be separated semi-continuously or continuously during the enantioselective hydrolysis.
  • the unreacted diester (Formula 5) may be isolated from the organic phase and racemized to yield the racemic substrate (Formula 4).
  • the resulting racemate (Formula 4) may be recycled or combined with unconverted racemic substrate, which subsequently undergoes enzymatic conversion to Formula 3 as described above. Recycling the unreacted diester (Formula 5) increases the overall yield of the enantioselective hydrolysis above 50%, thereby increasing the atom economy of the method and lowering costs associated with disposal of the undesired enantiomers.
  • the treatment of the diester (Formula 5) with a base that is strong enough to abstract an acidic ⁇ -proton of the malonate moiety generally results in inversion of the stereogenic center and generation of the racemic substrate (Formula 4).
  • Useful bases include organic bases, such as alkoxides (e.g., sodium ethoxide), linear aliphatic amines, and cyclic amines, and inorganic bases, such as KOH, NaOH, NH 4 OH, and the like.
  • the reaction is carried out in a compatible solvent, including polar protic solvents, such as EtOH or aprotic polar solvents, such as MTBE. Reaction temperatures above RT typically improve the yield of the racemization process.
  • the substantially enantiopure dicarboxylic acid monoester (Formula 3) may be converted to an optically active ⁇ -amino acid (Formula 1) using at least three different methods.
  • the monoester (Formula 3) is hydrolyzed in the presence of an acid catalyst or a base catalyst to yield an optically-active cyano-substituted dicarboxylic acid (Formula 6) or corresponding salt.
  • the cyano moiety of the resulting dicarboxylic acid (or its salt) is reduced to yield an optically-active ⁇ -amino dicarboxylic acid (Formula 7) or a corresponding salt, which is subsequently decarboxylated by treatment with an acid, by heating, or both, to yield the desired optically active ⁇ -amino acid (Formula 1).
  • the cyano moiety may be reduced via reaction with H 2 in the presence of catalytic amounts of Raney nickel, palladium, platinum, and the like, or through reaction with a reducing agent, such as LiAlH 4 , BH 3 —Me 2 S, and the like.
  • Useful acids for the hydrolysis and decarboxylation reactions include mineral acids, such as HClO 4 , H 1 , H 2 SO 4 , HBr, HCl, and the like.
  • Useful base catalysts for the hydrolysis reaction include various alkali and alkaline earth metal hydroxides and oxides, including LiOH, NaOH, KOH, and the like.
  • the dicarboxylic acid monoester (Formula 3) undergoes reductive cyclization to form an optically-active cyclic 3-carboxy-pyrrolidin-2-one (Formula 2), which is subsequently treated with an acid to yield the desired enantiomerically-enriched ⁇ -amino acid (Formula 1).
  • the reductive cyclization may be carried out by reacting the monoester (Formula 3) with H 2 in the presence of catalytic amounts of Raney nickel, palladium, platinum and the like.
  • One or more acids may be used to hydrolyze and decarboxylate the resulting lactam acid (Formula 2), including mineral acids such as HClO 4 , H 1 , H 2 SO 4 , HBr, and HCl, and organic acids such as HOAc, TFA, p-TSA, and the like.
  • the concentration of the acids may range from about 1N to about 12 N, and the amount of the acids may range from about 1 eq to about 7 eq.
  • the hydrolysis and decarboxylation reactions may be carried out at a temperature of about RT or higher, or at a temperature of about 60° C. or higher, or at temperature in a range of about 60° C. to about 130° C.
  • the ester moiety of the dicarboxylic acid monoester (Formula 3) is first hydrolyzed to give the cyano-substituted dicarboxylic acid (Formula 6 or its salt) as described above.
  • the resulting dicarboxylic acid (or its salt) is subsequently decarboxylated to give an optically-active cyano-substituted carboxylic acid or its salt (Formula 8 in which R 5 is a hydrogen atom, though R 5 can also be C 1-12 alkyl, C 3-12 cycloalkyl, or aryl-C 1-6 alkyl as noted below).
  • the dicarboxylic acid monoester (Formula 3) may be first decarboxylated directly to a cyano substituted monoester (Formula 8) by heating the aqueous solution of the dicarboxylic acid monoester (as a salt) to a temperature of from about 50° C. to reflux.
  • Krapcho conditions DMSO/NaCl/water
  • the cyano moiety of the compound of formula 8 is subsequently reduced to give the optically active ⁇ -amino acid (Formula 1).
  • a number of other catalysts may be used to reduce the cyano moiety of the compounds of Formula 3, 6 and 8.
  • These include, without limitation, heterogeneous catalysts containing from about 0.1% to about 20%, and more typically, from about 1% to about 5%, by weight, of transition metals such as Ni, Pd, Pt, Rh, Re, Ru, and Ir, including oxides and combinations thereof, which are typically supported on various materials, including Al 2 O 3 , C, CaCO 3 , SrCO 3 , BaSO 4 , MgO, SiO 2 , TiO 2 , ZrO 2 , and the like.
  • Useful catalysts thus include palladium catalysts such as Pd/C, Pd/SrCO 3 , Pd/Al 2 O 3 , Pd/MgO, Pd/CaCO 3 , Pd/BaSO 4 , PdO, Pd black, PdCl 2 , and the like, containing from about 1% to about 5% Pd, based on weight.
  • Other useful catalysts include Rh/C, Ru/C, Re/C, PtO 2 , Rh/C, RuO 2 , and the like.
  • the catalytic reduction of the cyano moiety is typically carried out in the presence of one or more polar solvents, including without limitation, water, alcohols, ethers, esters and acids, such as MeOH, EtOH, IPA, THF, EtOAc, and HOAc.
  • the reaction may be carried out at temperatures ranging from about 5° C. to about 100° C., though reactions at RT are common.
  • the substrate-to-catalyst ratio may range from about 1:1 to about 1000:1, based on weight, and H 2 pressure may range from about atmospheric pressure, 0 psig, to about 1500 psig. More typically, the substrate-to-catalyst ratios range from about 4:1 to about 20: 1, and H 2 pressures range from about 25 psig to about 150 psig.
  • Isolation of the lactam acid reduces the carryover of water-soluble impurities into the final product mixture and permits higher reactant concentration (e.g., about 1 M to about 2 M) during hydrolysis and decarboxylation, thereby increasing process throughput.
  • reactant concentration e.g., about 1 M to about 2 M
  • direct decarboxylation by heating the aqueous solution of the dicarboxylic acid monoester (Formula 3) affords the cyanomonoester (Formula 8) in high enantiomeric purity.
  • This compound can be separated from the reaction medium by extraction in an organic solvent or by direct phase separation, ensuring efficient removal of inorganic impurities by the water phase. High reaction throughput and the avoidance of strongly acidic conditions are two advantages of this approach.
  • FIG. 2 illustrates a process for preparing cyano-substituted diesters (Formula 4), which may serve as substrates for the enzymatic enantioselective hydrolysis shown in FIG. 1 .
  • the process includes a crossed aldol condensation, which comprises reacting an unsymmetrical ketone or an aldehyde (Formula 17) with a malonic acid diester (Formula 18) in the presence of catalytic amounts of a base to yield an ⁇ , ⁇ -unsaturated malonic acid diester (Formula 19) in which R 1 , R 2 , R 3 , and R 4 are as defined above in connection with Formula 1.
  • any base capable of generating an enolate ion from the malonic acid diester may be used, including secondary amines, such as di-n-propylamine, di-1-propylamine, pyrrolidine, etc., and their salts.
  • the reaction may include a carboxylic acid, such as HOAc, to neutralize the product and to minimize enolization of the unsymmetrical ketone or aldehyde (Formula 17).
  • Reactions involving unsymmetrical ketones may also employ Lewis acids, such as titanium tetrachloride, zinc chloride, zinc acetate, and the like to facilitate reaction.
  • the reaction is typically run in a hydrocarbon solvent under reflux conditions.
  • Useful solvents include hexane, heptane, cyclohexane, toluene, methyl t-butyl ether, and the like, with azeotropic removal of water.
  • a cyanide source such as HCN, acetone cyanohydrin, an alkali metal cyanide (NaCN, KCN, etc.), or an alkaline earth metal cyanide (magnesium cyanide, etc.
  • a cyanide source undergoes conjugate addition to the ⁇ -carbon of the ⁇ , ⁇ -unsaturated malonic acid diester (Formula 19).
  • the reaction is typically carried out in one or more polar protic solvents, including EtOH, MeOH, n-propanol, isopropanol, or polar aprotic solvents, such as DMSO, and the like.
  • polar protic solvents including EtOH, MeOH, n-propanol, isopropanol, or polar aprotic solvents, such as DMSO, and the like.
  • polar protic solvents such as EtOH, MeOH, n-propanol, isopropanol, or polar aprotic solvents, such
  • the desired (S)- or (R)-enantiomers of any of the compounds disclosed herein may be further enriched through classical resolution, chiral chromatography, or recrystallization.
  • the optically active ⁇ -amino acids (Formula 1 or Formula 9) may be reacted with an enantiomerically-pure compound (e.g., acid or base) to yield a pair of diastereoisomers, each composed of a single enantiomer, which are separated via, say, fractional recrystallization or chromatography.
  • the desired enantiomer is subsequently regenerated from the appropriate diastereoisomer.
  • the desired enantiomer often may be further enriched by recrystallization in a suitable solvent when it is it available in sufficient quantity (e.g., typically not much less than about 85% ee, and in some cases, not much less than about 90% ee).
  • stereoisomers As described throughout the specification, many of the disclosed compounds have stereoisomers. Some of these compounds may exist as single enantiomers (enantiopure compounds) or mixtures of enantiomers (enriched and racemic samples), which depending on the relative excess of one enantiomer over another in a sample, may exhibit optical activity. Such stereoisomers, which are non-superimposable mirror images, possess a stereogenic axis or one or more stereogenic centers (i.e., chirality). Other disclosed compounds may be stereoisomers that are not mirror images. Such stereoisomers, which are known as diastereoisomers, may be chiral or achiral (contain no stereogenic centers).
  • the scope of the present invention generally includes the reference compound and its stereoisomers, whether they are each pure (e.g., enantiopure) or mixtures (e.g., enantiomerically enriched or racemic).
  • the compounds may also contain a keto or oxime group, so that tautomerism may occur.
  • the present invention generally includes tautomeric forms, whether they are each pure or mixtures.
  • salts include, without limitation, acid addition salts (including diacids) and base salts.
  • Pharmaceutically acceptable acid addition salts include nontoxic salts derived from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, hydrofluoric, phosphorous, and the like, as well nontoxic salts derived from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc.
  • Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate, and the like.
  • Pharmaceutically acceptable base salts include nontoxic salts derived from bases, including metal cations, such as an alkali or alkaline earth metal cation, as well as amines.
  • suitable metal cations include, without limitation, sodium cations (Na + ), potassium cations (K + ), magnesium cations (Mg 2+ ), calcium cations (Ca 2+ ), and the like.
  • suitable amines include, without limitation, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, procaine, and t-butyl amine.
  • Disclosed and claimed compounds may exist in both unsolvated and solvated forms and as other types of complexes besides salts.
  • Useful complexes include clathrates or compound-host inclusion complexes where the compound and host are present in stoichiometric or non-stoichiometric amounts.
  • Useful complexes may also contain two or more organic, inorganic, or organic and inorganic components in stoichiometric or non-stoichiometric amounts.
  • the resulting complexes may be ionized, partially ionized, or non-ionized.
  • solvates also include hydrates and solvates in which the crystallization solvent may be isotopically substituted, e.g. D 2 O, d 6 -acetone, d6-DMSO, etc.
  • references to an unsolvated form of a compound also include the corresponding solvated or hydrated form of the compound.
  • the disclosed compounds also include all pharmaceutically acceptable isotopic variations, in which at least one atom is replaced by an atom having the same atomic number, but an atomic mass different from the atomic mass usually found in nature.
  • isotopes suitable for inclusion in the disclosed compounds include, without limitation, isotopes of hydrogen, such as 2 H and 3 H; isotopes of carbon, such as 13 C and 14 C; isotopes of nitrogen, such as 15 N; isotopes of oxygen, such as 17 O and 18 O; isotopes of phosphorus, such as 31 P and 32 P; isotopes of sulfur, such as 35 S; isotopes of fluorine, such as 18 F; and isotopes of chlorine, such as 36 Cl.
  • isotopic variations e.g., deuterium, 2H
  • isotopic variations of the disclosed compounds may incorporate a radioactive isotope (e.g., tritium, 3 H, or 14 C), which may be useful in drug and/or substrate tissue distribution studies.
  • a radioactive isotope e.g., tritium, 3 H, or 14 C
  • Enzyme screening was carried out using a 96-well plate, which is described in D. Yazbeck et al., Synth. Catal. 345:524-32 (2003), the complete disclosure of which is herein incorporated by reference for all purposes. All enzymes used in the screening plate (see Table 2) were obtained from commercial enzyme suppliers including Amano (Nagoya, Japan), Roche (Basel, Switzerland), Novo Nordisk (Bagsvaerd, Denmark), Altus Biologics Inc. (Cambridge, Mass.), Biocatalytics (Pasadena, Calif.), Toyobo (Osaka, Japan), Sigma-Aldrich (St. Louis, Mo.) and Fluka (Buchs, Switzerland).
  • Regular 13 C NMR spectra were acquired using 30-degree tip angle pulses, 2.0 s recycle delay, and 2048 scans at a resolution of 1 Hz/point.
  • Spectral width was typically 25 KHz from +235 to ⁇ 15 ppm (Reference TMS at 0 ppm).
  • Proton decoupling was applied continuously and 1 Hz line broadening was applied during processing. Typical acquisition time was 102 min.
  • Mass Spectrometry was performed on a HEWLETT PACKARD 1100MSD using HP Chemstation Plus Software.
  • the LC was equipped with an Agilent 1100 quaternary LC system and an Agilent liquid handler as an autosampler. Data were acquired under electron spray ionization with ACN/water (containing 0.1% formic acid) as the solvent (10% ACN to 90%, 7 min). Temperatures: probe was 350° C., source was 150° C. Corona discharge was 3000 V for positive ion and 3000 V for negative ion.
  • HPLC High Performance Liquid Chromatography
  • GC Gas Chromatography
  • Enzyme screening was carried out using a screening kit comprised of individual enzymes deposited in separate wells of a 96-well plate, which was prepared in advance in accordance with a method described in D. Yazbeck et al., Synth. Catal. 345:524-32 (2003). Each of the wells had an empty volume of 0.3 ml (shallow well plate).
  • One well of the 96-well plate contained only phosphate buffer (10 ⁇ L, 0.1 M, pH 7.2), another well contained only ACN (10 ⁇ L), and each of the remaining wells contained one of the 94 enzymes listed in Table 2 (10 ⁇ L, 100 mg/mL). Prior to use, the screening kit was removed from storage at ⁇ 80° C.
  • Potassium phosphate buffer (85 ⁇ L, 0.1 M, pH 7.2) was dispensed into each of the wells using a multi-channel pipette.
  • Concentrated substrate (Formula 20, 5 ⁇ L) was subsequently added to each well via a multi-channel pipette and the 96 reaction mixtures were incubated at 30° C. and 750 rpm. The reactions were quenched and sampled after 24 h by transferring each of the reaction mixtures into separate wells of a second 96-well plate. Each of the wells had an empty volume of 2 mL (deep well plate) and contained EtOAc (1 mL) and HCl (1N, 100 ⁇ L).
  • each well was mixed by aspirating the well contents with a pipette.
  • the second plate was centrifuged and 100 ⁇ L of the organic supernatant was transferred from each well into separate wells of a third 96-well plate (shallow plate).
  • the wells of the third plate were subsequently sealed using a penetrable mat cover. Once the wells were sealed, the third plate was transferred to a GC system for determination of optical purity (ee).
  • Table 3 lists enzyme, trade name, supplier, and E value for some of the enzymes that were screened.
  • the E value may be interpreted as the relative reactivity of a pair of enantiomers (substrates).
  • the E values listed in Table 3 were calculated from HPLC data (fractional conversion, ⁇ , and ee) using a computer program called Ee2, which is available from the University of Graz.
  • Ee2 which is available from the University of Graz.
  • enzymes exhibiting S-selectivity and an E value of about 35 or greater are suitable for scale-up.
  • the reaction mixture Upon reaching a conversion of about 40-45% (e.g., after about 24 h) the reaction mixture was transferred to a separatory funnel. The aqueous mixture was extracted with heptane (205 L). EtOH (absolute) was added (up to about 5% v/v) to disrupt a light emulsion that formed, and the aqueous and organic layers were separated. The extraction step was repeated twice, and the aqueous layer containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23) may be further concentrated under vacuum (e.g., 25-50% of its original volume).
  • a reactor (3.92 L) equipped with overhead stirring is charged with calcium acetate buffer (1.47 L, 100 mM, pH 7.0) and (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20, 1 kg).
  • the mixture is stirred at 1100 RPM for 5 min and KOH (5 M) is added to adjust the pH to 7.0.
  • LIPOLASE® 100L, type EX (75 mL) is added and the resulting mixture is titrated with KOH (5 M) during hydrolysis to maintain a pH of 7.0.
  • the extent of reaction is monitored by HPLC (C 18 column, 4.6 mm ⁇ 150 mm, detection at 200 nm).
  • the reaction mixture Upon reaching a conversion of about 42% to 45% (e.g., after about 20-25 h) the reaction mixture is transferred to a separatory funnel.
  • the aqueous mixture is extracted with hexane (100% v/v).
  • EtOH absolute
  • EtOH absolute
  • the extraction step is repeated twice to obtain an aqueous layer containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23), which may be used in subsequent transformations without isolation.
  • a vessel was charged with an aqueous solution containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23, 411 L from Example 2).
  • Raney Nickel (50% aq solution, Sigma-Aldrich) was added to the mixture, and hydrogen gas was introduced into the vessel over a 20 h period to maintain a pressure of 50 psig in the vessel headspace throughout reaction.
  • the hydrogenation reaction was monitored by H 2 uptake and HPLC analysis (C 18 column, 4.6 mm ⁇ 150 mm, detection at 200 nm) of the vessel contents.
  • the aqueous mixture was filtered to remove the Raney Ni catalyst.
  • the pH of the concentrated solution was adjusted to 3.0 using 37% HCl (about 14 L).
  • optical purity of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid was determined via a derivatization method.
  • a sample of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid was esterified with EtOH in the presence of a catalytic amount of dry HCl in dioxane at 70° C.
  • the resulting lactam ester was analyzed by HPLC (CHIRALPAK AD-H, 4.6 mm ⁇ 250 mm) using a mobile phase of hexane and EtOH (95:5), a flow rate of 1.0 mL/min, injection volume of 10 ⁇ L, column temperature of 35° C., and detection at 200 nm.
  • pregabalin The optical purity of pregabalin was analyzed via a derivatization method.
  • a sample of pregabalin was derivatized with Marfey's reagent (1-fluoro-24-dinitrophenyl-5-L-alanine amide) and then analyzed by HPLC (LUNA C 18 (2) column, 0.46 mm ⁇ 150 mm, 3 ⁇ m) using a mobile phase of aqueous NaPO 4 (20 nM, pH 2.0) and ACN (90:10 for 10 min, 10:90 for 3 min, 90:10 for 5 min), a flow rate of 1.2 mL/min, an injection volume of 10 ⁇ L, column temperature of 35° C., and detection at 200 nm.
  • a reactor (16000 L) equipped with overhead stirring is charged with calcium acetate (254 kg), deionized water (1892.7 kg) and LIPOZYME® TL 100 L (food grade LIPOLASE®, 983.7 kg).
  • (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20, 9000 kg, 85% purity assay) is charged and the mixture is stirred for 24 h.
  • NaOH (2068 kg of a 30% solution) is added over the course of the reaction to maintain the pH at 7.0.
  • the extent of reaction is monitored by HPLC (C 18 column, 4.6 mm ⁇ 150 mm, detection at 200 nm).
  • a reactor (16000 L) equipped with overhead stirring is charged with the final aqueous solution from Example 10 (9698.6 L, containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid sodium salt, Formula 23), NaCl (630 kg) and toluene (900 L).
  • the mixture is stirred for 2 h under refluxing conditions (75-85° C.).
  • the stirring is stopped; the organic phase is immediately separated and the aqueous phase is washed twice with toluene (900 L).
  • the organic layers, which contain (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24) are combined and concentrated.
  • the ethyl ester (Formula 24) is subsequently hydrolyzed in accordance with Example 12.
  • a hydrogenator (12000 L) is charged with water (942.1 L) and with the reaction mixture from Example 12, which contains (S)-3-cyano-5-methyl-hexanoic acid potassium salt (Formula 26,4122.9 L).
  • a Raney nickel suspension (219.6 kg, 50% w/w in H 2 O) is added. The hydrogenation is conducted under 50 psig at 35° C. After 6 h, the Raney nickel is filtered and the resulting filtrate is transferred to a reactor (16000 L) for crystallization. After adding H 2 O (1098 L), the pH of the solution is adjusted to 7.0-7.5 using HOAc (864.7 kg). The resulting precipitate is filtered and washed once with H 2 O (549 L) and twice with IPA (2,586 L each).
  • the solid is recrystallized with IPA (12296 L) and H 2 O (6148 L). The mixture is heated to 70° C. and subsequently cooled to 4° C. After 5-10 h, the crystalline solid is filtered, washed with IPA (5724 L), and dried in a vacuum oven at 45° C. for 24 h to give pregabalin as a white crystalline solid (1431 kg, 30.0% overall yield, 99.5% purity and 99.75% ee).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Veterinary Medicine (AREA)
  • Pain & Pain Management (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medicinal Chemistry (AREA)
  • Neurosurgery (AREA)
  • Neurology (AREA)
  • Biomedical Technology (AREA)
  • Public Health (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Pyrrole Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Low-Molecular Organic Synthesis Reactions Using Catalysts (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Steroid Compounds (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Nitrogen And Oxygen As The Only Ring Hetero Atoms (AREA)

Abstract

Materials and methods for preparing (S)-(+)-3-aminomethyl-5-methyl-hexanoic acid and structurally related compounds via enzymatic kinetic resolution are disclosed.

Description

    CROSS REFERENCE TO RELATED APPLICATIONS
  • This application claims the benefit of U.S. Provisional Application No. 60/581,671, filed Jun. 21, 2004, and U.S. Provisional Application No. 60/629,034, filed Nov. 18, 2004, the complete disclosures of which are herein incorporated by reference.
    • BACKGROUND OF THE INVENTION
  • 1. Field of the Invention
  • This invention relates to methods and materials for preparing enantiomerically-enriched γ-amino acids via enzymatic kinetic resolution, and is particularly useful for preparing γ-amino acids that exhibit binding affinity to the human α2δ calcium channel subunit, including pregabalin and related compounds.
  • 2. Discussion
  • Pregabalin, (S)-(+)-3-aminomethyl-5-methyl-hexanoic acid, is related to the endogenous inhibitory neurotransmitter γ-aminobutyric acid (GABA), which is involved in the regulation of brain neuronal activity. Pregabalin exhibits anti-seizure activity, as discussed in U.S. Pat. No. 5,563,175 to R. B. Silverman et al., and is thought to be useful for treating, among other conditions, pain, physiological conditions associated with psychomotor stimulants, inflammation, gastrointestinal damage, alcoholism, insomnia, and various psychiatric disorders, including mania and bipolar disorder. See, respectively, U.S. Pat. No. 6,242,488 to L. Bueno et al., U.S. Pat. No. 6,326,374 to L. Magnus & C. A. Segal, and U.S. Pat. No. 6,001,876 to L. Singh; U.S. Pat. No. 6,194,459 to H. C. Akunne et al.; U.S. Pat. No. 6,329,429 to D. Schrier et al.; U.S. Pat. No. 6,127,418 to L. Bueno et al.; U.S. Pat. No. 6,426,368 to L. Bueno et al.; U.S. Pat. No. 6,306,910 to L. Magnus & C. A. Segal; and U.S. Pat. No. 6,359,005 to A. C. Pande, which are herein incorporated by reference in their entirety and for all purposes.
  • Pregabalin has been prepared in various ways. Typically, a racemic mixture of 3-aminomethyl-5-methyl-hexanoic acid is synthesized and subsequently resolved into its R- and S-enantiomers. Such methods may employ an azide intermediate, a malonate intermediate, or Hofman synthesis. See, respectively, U.S. Pat. No. 5,563,175 to R. B. Silverman et al.; U.S. Pat. Nos. 6,046,353, 5,840,956, and 5,637,767 to T. M. Grote et al.; and U.S. Pat. Nos. 5,629,447 and 5,616,793 to B. K. Huckabee & D. M. Sobieray, which are herein incorporated by reference in their entirety and for all purposes. In each of these methods, the racemate is reacted with a chiral acid (a resolving agent) to form a pair of diastereoisomeric salts, which are separated by known techniques, such as fractional crystallization and chromatography. These methods thus involve significant processing beyond the preparation of the racemate, which along with the resolving agent, adds to production costs. Moreover, the undesired R-enantiomer is frequently discarded since it cannot be efficiently recycled, thereby reducing the effective throughput of the process by 50%.
  • Pregabalin has also been synthesized directly using a chiral auxiliary, (4R,5S)-4-methyl-5-phenyl-2-oxazolidinone. See, e.g., U.S. Pat. Nos. 6,359,169, 6,028,214, 5,847,151, 5,710,304, 5,684,189, 5,608,090, and 5,599,973, all to R. B. Silverman et al, which are herein incorporated by reference in their entirety and for all purposes. Although these methods provide pregabalin in high enantiomeric purity, they are less desirable for large-scale synthesis because they employ comparatively costly reagents (e.g., the chiral auxiliary) that are difficult to handle, as well as special cryogenic equipment to reach required operating temperatures, which can be as low as −78° C.
  • A recently published U.S. patent application discusses a method of making pregabalin via asymmetric hydrogenation of a cyano-substituted olefin to produce a chiral cyano precursor of (S)-3-aminomethyl-5-methylhexanoic acid. See commonly assigned U.S. Patent Application No. 2003/0212290 A1 to Burk et al., published Nov. 13, 2003, which is herein incorporated by reference in its entirety for all purposes. The cyano precursor is subsequently reduced to give pregabalin. The asymmetric hydrogenation employs a chiral catalyst that is comprised of a transition metal bound to a bisphosphine ligand, such as (R,R)-Me-DUPHOS. The method results in substantial enrichment of pregabalin over (R)-3-(aminomethyl)-5-methylhexanoic acid.
  • The method discussed in U.S. Patent Application No. 2003/0212290 A1 represents a commercially viable method for preparing pregabalin, but further improvements would be desirable for various reasons. For example, bisphosphine ligands, including the proprietary ligand (R,R)-Me-DUPHOS, are often difficult to prepare because they possess two chiral centers, which adds to their cost. Furthermore, asymmetric hydrogenation requires the use of special equipment capable of handling H2, which adds to capital costs.
    • SUMMARY OF THE INVENTION
  • The present invention provides materials and methods for preparing enantiomerically enriched γ-amino acids (Formula 1) such as pregabalin (Formula 9). The method of the present invention involves a kinetic resolution of a racemic cyano diester intermediate (Formula 4 or Formula 12) using an enzyme that is adapted to enantioselectively hydrolyze an ester moiety of the intermediate. The resulting dicarboxylic acid monoester (Formula 3 or Formula 11), which is substantially enantiopure, undergoes further reaction to yield the desired enantiomerically-enriched γ-amino acids (Formula 1 or Formula 9). The unreacted enantiomer (Formula 5 or Formula 13) from the kinetic resolution can be reused in the enzymatic resolution following racemization, thereby improving overall yield.
  • The claimed method offers significant advantages over existing processes for preparing enantiomerically-enriched γ-amino acids (Formula 1 and Formula 9). For example, the optically-active γ-amino acids can be prepared without using chiral auxiliaries or proprietary hydrogenation catalysts, which should lead to lower unit costs. Since enzymatic processes can be carried out at room temperature and at atmospheric pressure, the claimed methods should help minimize scheduling conflicts arising from the use of specialized equipment capable of handling high pressures and low temperatures. As noted in the examples, the present invention can be used to prepare pregabalin starting from a racemic cyano-substituted diester (Formula 12) in good yield (26% to 31%) after a single batch recycle of the unreacted enantiomer (Formula 13). This translates into about a 50% savings in cost of goods over the malonate method described above.
  • One aspect of the present invention provides a method of making a compound of
    Figure US20050283023A1-20051222-C00001
    or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, in which
      • R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl,
        the method comprising:
      • (a) reacting a compound of Formula 2,
        Figure US20050283023A1-20051222-C00002
        or a salt thereof, with an acid and water to yield the compound of Formula 1 or a salt thereof; and
      • (b) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 2 are as defined in Formula 1.
  • Another aspect of the present invention provides a method of making a compound of Formula 1, above, the method comprising:
      • (a) reducing a cyano moiety of a compound of Formula 6,
        Figure US20050283023A1-20051222-C00003
        or a salt thereof, to yield a compound of Formula 7,
        Figure US20050283023A1-20051222-C00004
        or a salt thereof;
      • (b) decarboxylating the compound of Formula 7 or a salt thereof to yield the compound of Formula 1 or a salt thereof; and
      • (c) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 6 and in Formula 7 are as defined above in Formula 1.
  • The compound of Formula 6, above, may be prepared by hydrolyzing a compound of Formula 3,
    Figure US20050283023A1-20051222-C00005
    or a salt thereof, wherein R1 and R2 in Formula 3 are as defined above in Formula 1, and R3 is C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
  • An additional aspect of the present invention provides method of making a compound of Formula 1, above, the method comprising:
      • (a) reducing a cyano moiety of a compound of Formula 8,
        Figure US20050283023A1-20051222-C00006
        or a salt thereof, to yield the compound of Formula 1 or a salt thereof; and
      • (b) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 8 are as defined above in Formula 1, and R5 in Formula 8 is hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
  • The compound of Formula 8 may be prepared by decarboxylating a compound of Formula 3, above, or a salt thereof, or by hydrolyzing and decarboxylating the compound of Formula 3 or a salt thereof, to yield the compound of Formula 8 or a salt thereof.
  • A further aspect of the present invention provides a method of making the compound of Formula 3, above, or a salt thereof, the method comprising:
      • (a) contacting a compound of Formula 4,
        Figure US20050283023A1-20051222-C00007
        with an enzyme to yield the compound of Formula 3 and a compound of Formula 5,
        Figure US20050283023A1-20051222-C00008
        wherein the enzyme is adapted to enantioselectively hydrolyze the compound of Formula 4 to the compound of Formula 3 or a salt thereof;
      • (b) isolating the compound of Formula 3 or a salt thereof; and
      • (c) optionally racemizing the compound of Formula 5 to yield the compound of Formula 4, wherein R1, R2, and R3 in Formula 4 and Formula 5 are as defined above in Formula 1 and Formula 3; and R4 in Formula 4 and Formula 5 is the same as or different than R3 and is C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
  • Any number of enzymes may be used to enantioselectively hydrolyze the compound of Formula 4 to the compound of Formula 3 or a salt thereof. Useful enzymes include lipases, such as those derived from Thermomyces lanuginosus.
  • Another aspect of the present invention provides compounds represented by Formula 2, above, including complexes, salts, solvates or hydrates thereof, provided that when one of the substituents represented by R1 or R2 in Formula 2 is hydrogen, the other substituent is not C1-3 alkyl or C5 alkyl.
  • An additional aspect of the present invention provides compounds of Formula 27,
    Figure US20050283023A1-20051222-C00009
    including complexes, salts, solvates or hydrates thereof, wherein
      • R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl, provided that when one of the substituents represented by R1 or R2 is a hydrogen atom, the other substituent is not methyl; and
      • R5 and R6 are independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl, provided that R5 and R6 are different if not hydrogen atoms.
  • Compounds of Formula 27 include those represented by Formula 3, Formula 4, Formula 5, Formula 6, and Formula 7, above, including their complexes, salts, solvates or hydrates. Useful compounds of Formula 2-7 and 27 include those in which R1 is a hydrogen atom and R is isobutyl.
  • A further aspect of the present invention provides a method of making a compound of Formula 9,
    Figure US20050283023A1-20051222-C00010
    or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
      • (a) reacting a compound of Formula 10,
        Figure US20050283023A1-20051222-C00011
        or a salt thereof, with an acid and water to yield the compound of Formula 9 or a salt thereof; and
      • (b) optionally converting the compound of Formula 9 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate.
  • The compound of Formula 10 or a salt thereof may be prepared by reducing a cyano moiety of a compound of Formula 11,
    Figure US20050283023A1-20051222-C00012
    or a salt thereof, wherein R3 is as defined above in Formula 3.
  • Another aspect of the present invention provides a method of making a compound of Formula 9, above, or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
      • (a) reducing a cyano moiety of a compound of Formula 14,
        Figure US20050283023A1-20051222-C00013
        or a salt thereof, to yield a compound of Formula 15,
        Figure US20050283023A1-20051222-C00014
        or a salt thereof;
      • (b) decarboxylating the compound of Formula 15 or a salt thereof to yield the compound of Formula 9 or a salt thereof; and
      • (c) optionally converting the compound of Formula 9 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate.
  • The compound of Formula 14, above, may be prepared by hydrolyzing a compound of Formula 11,
    Figure US20050283023A1-20051222-C00015
    or salt thereof, wherein R3 in Formula 11 is as defined above in Formula 3.
  • An additional aspect of the present invention provides a method of making a compound of Formula 9, above, or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, the method comprising:
      • (a) reducing a cyano moiety of a compound of Formula 16,
        Figure US20050283023A1-20051222-C00016
        or a salt thereof, to yield the compound of Formula 9 or a salt thereof; and
      • (b) optionally converting the compound of Formula 9 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R5 in Formula 16 is as defined above in Formula 8.
  • The compound of Formula 16 may be prepared by decarboxylating (e.g., by heating) the compound of Formula 11, above, or a salt thereof, or by hydrolyzing and decarboxylating the compound of Formula 11 or a salt thereof.
  • A further aspect of the present invention provides a method of making the compound of Formula 11, above, or a salt thereof, the method comprising:
      • (a) contacting a compound of Formula 12,
        Figure US20050283023A1-20051222-C00017
        with an enzyme to yield the compound of Formula 11 and a compound of Formula 13,
        Figure US20050283023A1-20051222-C00018
        wherein the enzyme is adapted to enantioselectively hydrolyze the compound of Formula 12 to the compound of Formula 11 or a salt thereof;
      • (b) isolating the compound of Formula 11 or its salts thereof; and
      • (c) optionally racemizing the compound of Formula 13 to yield the compound of Formula 12, wherein
      • R3 in Formula 12 and Formula 13 is as defined above in Formula 3; and
      • R4 in Formula 12 and Formula 13 is as defined above in Formula 4 and 5.
  • In the method for preparing the compound of Formula 11, the corresponding salts of the compound of Formula 11 include those selected from alkali metal salts, such as potassium salt; primary amine salts, such as a t-butyl amine salt; and secondary amine salts. Furthermore, useful enzymes include lipases, such as those derived from Thermomyces lanuginosus.
  • Another aspect of the present invention provides a compound selected from:
    • 3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
    • (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
    • (2S,3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
    • (2R,3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
    • 3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester,
    • (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester,
    • 4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid,
    • (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid,
    • 3-cyano-2-carboxy-5-methyl-hexanoic acid,
    • (S)-3-cyano-2-carboxy-5-methyl-hexanoic acid,
    • 3-aminomethyl-2-carboxy-5-methyl-hexanoic acid, and
    • (S)-3-aminomethyl-2-carboxy-5-methyl-hexanoic acid,
      • including complexes, salts, solvates, and hydrates thereof and opposite enantiomers thereof.
  • The present invention includes all complexes and salts, whether pharmaceutically acceptable or not, solvates, hydrates, and polymorphic forms of the disclosed compounds. Certain compounds may contain an alkenyl or cyclic group, so that cisltrans (or Z/E) stereoisomers are possible, or may contain a keto or oxime group, so that tautomerism may occur. In such cases, the present invention generally includes all Z/E isomers and tautomeric forms, whether they are pure, substantially pure, or mixtures.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1 depicts a scheme for preparing enantiomerically-enriched γ-amino acids (Formula 1).
  • FIG. 2 depicts a scheme for preparing cyano-substituted diesters (Formula 4).
    • DETAILED DESCRIPTION
  • Definitions and Abbreviations
  • Unless otherwise indicated, this disclosure uses definitions provided below. Some of the definitions and formulae may include a dash (“−”) to indicate a bond between atoms or a point of attachment to a named or unnamed atom or group of atoms. Other definitions and formulae may include an equal sign (“=”) or an identity symbol (“≡”) to indicate a double bond or a triple bond, respectively. Certain formulae may also include one or more asterisks (“*”) to indicate stereogenic (asymmetric or chiral) centers, although the absence of an asterisk does not indicate that the compound lacks a stereocenter. Such formulae may refer to the racemate or to individual enantiomers or to individual diastereomers, which may or may not be pure or substantially pure.
  • “Substituted” groups are those in which one or more hydrogen atoms have been replaced with one or more non-hydrogen groups, provided that valence requirements are met and that a chemically stable compound results from the substitution.
  • “About” or “approximately,” when used in connection with a measurable numerical variable, refers to the indicated value of the variable and to all values of the variable that are within the experimental error of the indicated value (e.g., within the 95% confidence interval for the mean) or within ±10 percent of the indicated value, whichever is greater.
  • “Alkyl” refers to straight chain and branched saturated hydrocarbon groups, generally having a specified number of carbon atoms (i.e., C1-6 alkyl refers to an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms and C1-12 alkyl refers to an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms). Examples of alkyl groups include, without limitation, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, pent-1-yl, pent-2-yl, pent-3-yl, 3-methylbut-1-yl, 3-methylbut-2-yl, 2-methylbut-2-yl, 2,2,2-trimethyleth-1-yl, n-hexyl, and the like.
  • “Alkenyl” refers to straight chain and branched hydrocarbon groups having one or more unsaturated carbon-carbon bonds, and generally having a specified number of carbon atoms. Examples of alkenyl groups include, without limitation, ethenyl, 1-propen-1-yl, 1-propen-2-yl, 2-propen-1-yl, 1-buten-1-yl, 1-buten-2-yl, 3-buten-1-yl, 3-buten-2-yl, 2-buten-1-yl, 2-buten-2-yl, 2-methyl-1-propen-1-yl, 2-methyl-2-propen-1-yl, 1,3-butadien-1-yl, 1,3-butadien-2-yl, and the like.
  • “Alkynyl” refers to straight chain or branched hydrocarbon groups having one or more triple carbon-carbon bonds, and generally having a specified number of carbon atoms. Examples of alkynyl groups include, without limitation, ethynyl, 1-propyn-1-yl, 2-propyn-1-yl, 1-butyn-1-yl, 3-butyn-1-yl, 3-butyn-2-yl, 2-butyn-1-yl, and the like.
  • “Alkanoyl” and “alkanoylamino” refer, respectively, to alkyl-C(O)— and alkyl-C(O)—NH—, where alkyl is defined above, and generally includes a specified number of carbon atoms, including the carbonyl carbon. Examples of alkanoyl groups include, without limitation, formyl, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, and the like.
  • “Alkenoyl” and “alkynoyl” refer, respectively, to alkenyl-C(O)— and alkynyl-C(O)—, where alkenyl and alkynyl are defined above. References to alkenoyl and alkynoyl generally include a specified number of carbon atoms, excluding the carbonyl carbon. Examples of alkenoyl groups include, without limitation, propenoyl, 2-methylpropenoyl, 2-butenoyl, 3-butenoyl, 2-methyl-2-butenoyl, 2-methyl-3-butenoyl, 3-methyl-3-butenoyl, 2-pentenoyl, 3-pentenoyl, 4-pentenoyl, and the like. Examples of alkynoyl groups include, without limitation, propynoyl, 2-butynoyl, 3-butynoyl, 2-pentynoyl, 3-pentynoyl, 4-pentynoyl, and the like.
  • “Alkoxy,” “alkoxycarbonyl,” and “alkoxycarbonylamino,” refer, respectively, to alkyl-O—, alkenyl-O, and alkynyl-O; to alkyl-O—C(O)—, alkenyl-O—C(O)—, alkynyl-O—C(O)—; and to alkyl-O—C(O)—NH—, alkenyl-O—C(O)—NH—, and alkynyl-O—C(O)—NH—, where alkyl, alkenyl, and alkynyl are defined above. Examples of alkoxy groups include, without limitation, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, n-pentoxy, s-pentoxy, and the like. Examples of alkoxycarbonyl groups include, without limitation, methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, i-propoxycarbonyl, n-butoxycarbonyl, s-butoxycarbonyl, t-butoxycarbonyl, n-pentoxycarbonyl, s-pentoxycarbonyl, and the like.
  • “Alkylamino,” “alkylaminocarbonyl,” “di alkylaminocarbonyl,” “alkylsulfonyl” “sulfonylaminoalkyl,” and “alkylsulfonylaminocarbonyl” refer, respectively, to alkyl-NH—, alkyl-NH—C(O)—, alkyl2-N—C(O)—, alkyl-S(O2)—, HS(O2)—NH-alkyl-, and alkyl-S(O)—NH—C(O)— where alkyl is defined above.
  • “Aminoalkyl” and “cyanoalkyl” refer, respectively, to NH2-alkyl and N≡C-alkyl, where alkyl is defined above.
  • “Halo,” “halogen” and “halogeno” may be used interchangeably, and refer to fluoro, chloro, bromo, and iodo.
  • “Haloalkyl,” “haloalkenyl,” “haloalkynyl,” “haloalkanoyl,” “haloalkenoyl,” “haloalkynoyl,” “haloalkoxy,” and “haloalkoxycarbonyl” refer, respectively, to alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, alkoxy, and alkoxycarbonyl groups substituted with one or more halogen atoms, where alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, alkoxy, and alkoxycarbonyl are defined above. Examples of haloalkyl groups include, without limitation, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, and the like.
  • “Hydroxyalkyl” and “oxoalkyl” refer, respectively, to HO-alkyl and O=alkyl, where alkyl is defined above. Examples of hydroxyalkyl and oxoalkyl groups, include, without limitation, hydroxymethyl, hydroxyethyl, 3-hydroxypropyl, oxomethyl, oxoethyl, 3-oxopropyl, and the like.
  • “Cycloalkyl” refers to saturated monocyclic and bicyclic hydrocarbon rings, generally having a specified number of carbon atoms that comprise the ring (i.e., C3-7 cycloalkyl refers to a cycloalkyl group having 3, 4, 5, 6 or 7 carbon atoms as ring members). The cycloalkyl may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements. Likewise, the cycloalkyl groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements. Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, alkoxy, alkoxycarbonyl, alkanoyl, and halo, as defined above, and hydroxy, mercapto, nitro, and amino.
  • Examples of monocyclic cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. Examples of bicyclic cycloalkyl groups include, without limitation, bicyclo[1.1.0]butyl, bicyclo[1.1.1]pentyl, bicyclo[2.1.0]pentyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.0]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.0]heptyl, bicyclo[3.1.1]heptyl, bicyclo[4.1.0]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.1.1]octyl, bicyclo[3.3.0]octyl, bicyclo[4.2.0]octyl, bicyclo[3.3.1]nonyl, bicyclo[4.2.1]nonyl, bicyclo[4.3.0]nonyl, bicyclo[3.3.2]decyl, bicyclo[4.2.2]decyl, bicyclo[4.3.1]decyl, bicyclo[4.4.0]decyl, bicyclo[3.3.3]undecyl, bicyclo[4.3.2]undecyl, bicyclo[4.3.3]dodecyl, and the like, which may be attached to a parent group or substrate at any of the ring atoms, unless such attachment would violate valence requirements.
  • “Cycloalkenyl” refers monocyclic and bicyclic hydrocarbon rings having one or more unsaturated carbon-carbon bonds and generally having a specified number of carbon atoms that comprise the ring (i.e., C3-7 cycloalkenyl refers to a cycloalkenyl group having 3, 4, 5, 6 or 7 carbon atoms as ring members). The cycloalkenyl may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements. Likewise, the cycloalkenyl groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements. Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, alkoxy, alkoxycarbonyl, alkanoyl, and halo, as defined above, and hydroxy, mercapto, nitro, and amino.
  • “Cycloalkanoyl” and “cycloalkenoyl” refer to cycloalkyl-C(O)— and cycloalkenyl-C(O)—, respectively, where cycloalkyl and cycloalkenyl are defined above. References to cycloalkanoyl and cycloalkenoyl generally include a specified number of carbon atoms, excluding the carbonyl carbon. Examples of cycloalkanoyl groups include, without limitation, cyclopropanoyl, cyclobutanoyl, cyclopentanoyl, cyclohexanoyl, cycloheptanoyl, 1-cyclobutenoyl, 2-cyclobutenoyl, 1-cyclopentenoyl, 2-cyclopentenoyl, 3-cyclopentenoyl, 1-cyclohexenoyl, 2-cyclohexenoyl, 3-cyclohexenoyl, and the like.
  • “Cycloalkoxy” and “cycloalkoxycarbonyl” refer, respectively, to cycloalkyl-O— and cycloalkenyl-O and to cycloalkyl-O—C(O)— and cycloalkenyl-O—C(O)—, where cycloalkyl and cycloalkenyl are defined above. References to cycloalkoxy and cycloalkoxycarbonyl generally include a specified number of carbon atoms, excluding the carbonyl carbon. Examples of cycloalkoxy groups include, without limitation, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexoxy, 1-cyclobutenoxy, 2-cyclobutenoxy, 1-cyclopentenoxy, 2-cyclopentenoxy, 3-cyclopentenoxy, 1-cyclohexenoxy, 2-cyclohexenoxy, 3-cyclohexenoxy, and the like. Examples of cycloalkoxycarbonyl groups include, without limitation, cyclopropoxycarbonyl, cyclobutoxycarbonyl, cyclopentoxycarbonyl, cyclohexoxycarbonyl, 1-cyclobutenoxycarbonyl, 2-cyclobutenoxycarbonyl, 1-cyclopentenoxycarbonyl, 2-cyclopentenoxycarbonyl, 3-cyclopentenoxycarbonyl, 1-cyclohexenoxycarbonyl, 2-cyclohexenoxycarbonyl, 3-cyclohexenoxycarbonyl, and the like.
  • “Aryl” and “arylene” refer to monovalent and divalent aromatic groups, respectively, including 5- and 6-membered monocyclic aromatic groups that contain 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Examples of monocyclic aryl groups include, without limitation, phenyl, pyrrolyl, furanyl, thiopheneyl, thiazolyl, isothiazolyl, imidazolyl, triazolyl, tetrazolyl, pyrazolyl, oxazolyl, isooxazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, and the like. Aryl and arylene groups also include bicyclic groups, tricyclic groups, etc., including fused 5- and 6-membered rings described above. Examples of multicyclic aryl groups include, without limitation, naphthyl, biphenyl, anthracenyl, pyrenyl, carbazolyl, benzoxazolyl, benzodioxazolyl, benzothiazolyl, benzoimidazolyl, benzothiopheneyl, quinolinyl, isoquinolinyl, indolyl, benzofuranyl, purinyl, indolizinyl, and the like. They aryl and arylene groups may be attached to a parent group or to a substrate at any ring atom, unless such attachment would violate valence requirements. Likewise, aryl and arylene groups may include one or more non-hydrogen substituents unless such substitution would violate valence requirements. Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, alkanoyl, cycloalkanoyl, cycloalkenoyl, alkoxycarbonyl, cycloalkoxycarbonyl, and halo, as defined above, and hydroxy, mercapto, nitro, amino, and alkylamino.
  • “Heterocycle” and “heterocyclyl” refer to saturated, partially unsaturated, or unsaturated monocyclic or bicyclic rings having from 5 to 7 or from 7 to 11 ring members, respectively. These groups have ring members made up of carbon atoms and from 1 to 4 heteroatoms that are independently nitrogen, oxygen or sulfur, and may include any bicyclic group in which any of the above-defined monocyclic heterocycles are fused to a benzene ring. The nitrogen and sulfur heteroatoms may optionally be oxidized. The heterocyclic ring may be attached to a parent group or to a substrate at any heteroatom or carbon atom unless such attachment would violate valence requirements. Likewise, any of the carbon or nitrogen ring members may include a non-hydrogen substituent unless such substitution would violate valence requirements. Useful substituents include, without limitation, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, alkanoyl, cycloalkanoyl, cycloalkenoyl, alkoxycarbonyl, cycloalkoxycarbonyl, and halo, as defined above, and hydroxy, mercapto, nitro, amino, and alkylamino.
  • Examples of heterocycles include, without limitation, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.
  • “Heteroaryl” and “heteroarylene” refer, respectively, to monovalent and divalent heterocycles or heterocyclyl groups, as defined above, which are aromatic. Heteroaryl and heteroarylene groups represent a subset of aryl and arylene groups, respectively.
  • “Arylalkyl” and “heteroarylalkyl” refer, respectively, to aryl-alkyl and heteroaryl-alkyl, where aryl, heteroaryl, and alkyl are defined above. Examples include, without limitation, benzyl, fluorenylmethyl, imidazol-2-yl-methyl, and the like.
  • “Arylalkanoyl,” “heteroarylalkanoyl,” “arylalkenoyl,” “heteroarylalkenoyl,” “arylalkynoyl,” and “heteroarylalkynoyl” refer, respectively, to aryl-alkanoyl, heteroaryl-alkanoyl, aryl-alkenoyl, heteroaryl-alkenoyl, aryl-alkynoyl, and heteroaryl-alkynoyl, where aryl, heteroaryl, alkanoyl, alkenoyl, and alkynoyl are defined above. Examples include, without limitation, benzoyl, benzylcarbonyl, fluorenoyl, fluorenylmethylcarbonyl, imidazol-2-oyl, imidazol-2-yl-methylcarbonyl, phenylethenecarbonyl, 1-phenylethenecarbonyl, 1-phenyl-propenecarbonyl, 2-phenyl-propenecarbonyl, 3-phenyl-propenecarbonyl, imidazol-2-yl-ethenecarbonyl, 1-(imidazol-2-yl)-ethenecarbonyl, 1-(imidazol-2-yl)-propenecarbonyl, 2-(imidazol-2-yl)-propenecarbonyl, 3-(imidazol-2-yl)-propenecarbonyl, phenylethynecarbonyl, phenylpropynecarbonyl, (imidazol-2-yl)-ethynecarbonyl, (imidazol-2-yl)-propynecarbonyl, and the like.
  • “Arylalkoxy” and “heteroarylalkoxy” refer, respectively, to aryl-alkoxy and heteroaryl-alkoxy, where aryl, heteroaryl, and alkoxy are defined above. Examples include, without limitation, benzyloxy, fluorenylmethyloxy, imidazol-2-yl-methyloxy, and the like.
  • “Aryloxy” and “heteroaryloxy” refer, respectively, to aryl-O— and heteroaryl-O—, where aryl and heteroaryl are defined above. Examples include, without limitation, phenoxy, imidazol-2-yloxy, and the like.
  • “Aryloxycarbonyl,” “heteroaryloxycarbonyl,” “arylalkoxycarbonyl,” and “heteroarylalkoxycarbonyl” refer, respectively, to aryloxy-C(O)—, heteroaryloxy-C(O)—, arylalkoxy-C(O)—, and heteroarylalkoxy-C(O)—, where aryloxy, heteroaryloxy, arylalkoxy, and heteroarylalkoxy are defined above. Examples include, without limitation, phenoxycarbonyl, imidazol-2-yloxycarbonyl, benzyloxycarbonyl, fluorenylmethyloxycarbonyl, imidazol-2-yl-methyloxycarbonyl, and the like.
  • “Leaving group” refers to any group that leaves a molecule during a fragmentation process, including substitution reactions, elimination reactions, and addition-elimination reactions. Leaving groups may be nucleofugal, in which the group leaves with a pair of electrons that formerly served as the bond between the leaving group and the molecule, or may be electrofugal, in which the group leaves without the pair of electrons. The ability of a nucleofugal leaving group to leave depends on its base strength, with the strongest bases being the poorest leaving groups. Common nucleofugal leaving groups include nitrogen (e.g., from diazonium salts); sulfonates, including alkylsulfonates (e.g., mesylate), fluoroalkylsulfonates (e.g., triflate, hexaflate, nonaflate, and tresylate), and arylsulfonates (e.g., tosylate, brosylate, closylate, and nosylate). Others include carbonates, halide ions, carboxylate anions, phenolate ions, and alkoxides. Some stronger bases, such as NH2 − and OH− can be made better leaving groups by treatment with an acid. Common electrofugal leaving groups include the proton, CO2, and metals.
  • “Enantiomeric excess” or “ee” is a measure, for a given sample, of the excess of one enantiomer over a racemic sample of a chiral compound and is expressed as a percentage. Enantiomeric excess is defined as 100×(er−1)/(er+1), where “er” is the ratio of the more abundant enantiomer to the less abundant enantiomer.
  • “Diastereomeric excess” or “de” is a measure, for a given sample, of the excess of one diastereomer over a sample having equal amounts of diastereomers and is expressed as a percentage. Diastereomeric excess is defined as 100×(dr−1)/(dr+1), where “dr” is the ratio of a more abundant diastereomer to a less abundant diastereomer.
  • “Stereoselective,” “enantioselective,” “diastereoselective,” and variants thereof, refer to a given process (e.g., ester hydrolysis, hydrogenation, hydroformylation, π-allyl palladium coupling, hydrosilation, hydrocyanation, olefin metathesis, hydroacylation, allylamine isomerization, etc.) that yields more of one stereoisomer, enantiomer, or diastereoisomer than of another, respectively.
  • “High level of stereoselectivity,” “high level of enantioselectivity,” “high level of diastereoselectivity,” and variants thereof, refer to a given process that yields products having an excess of one stereoisomer, enantiomer, or diastereoisomer, which comprises at least about 90% of the products. For a pair of enantiomers or diastereomers, a high level of enantioselectivity or diastereoselectivity would correspond to an ee or de of at least about 80%.
  • “Stereoisomerically enriched,” “enantiomerically enriched,” “diastereomerically enriched,” and variants thereof, refer, respectively, to a sample of a compound that has more of one stereoisomer, enantiomer or diastereomer than another. The degree of enrichment may be measured by % of total product, or for a pair of enantiomers or diastereomers, by ee or de.
  • “Substantially pure stereoisomer,” “substantially pure enantiomer,” “substantially pure diastereomer,” and variants thereof, refer, respectively, to a sample containing a stereoisomer, enantiomer, or diastereomer, which comprises at least about 95% of the sample. For pairs of enantiomers and diastereomers, a substantially pure enantiomer or diastereomer would correspond to samples having an ee or de of about 90% or greater.
  • A “pure stereoisomer,” “pure enantiomer,” “pure diastereomer,” and variants thereof, refer, respectively, to a sample containing a stereoisomer, enantiomer, or diastereomer, which comprises at least about 99.5% of the sample. For pairs of enantiomers and diastereomers, a pure enantiomer or pure diastereomer” would correspond to samples having an ee or de of about 99% or greater.
  • “Opposite enantiomer” refers to a molecule that is a non-superimposable mirror image of a reference molecule, which may be obtained by inverting all of the stereogenic centers of the reference molecule. For example, if the reference molecule has S absolute stereochemical configuration, then the opposite enantiomer has R absolute stereochemical configuration. Likewise, if the reference molecule has S,S absolute stereochemical configuration, then the opposite enantiomer has R,R stereochemical configuration, and so on.
  • “Stereoisomers” of a specified compound refer to the opposite enantiomer of the compound and to any diastereoisomers or geometric isomers (ZiE) of the compound. For example, if the specified compound has S,R,Z stereochemical configuration, its stereoisomers would include its opposite enantiomer having R,S,Z configuration, its diastereomers having S,S,Z configuration and R,R,Z configuration, and its geometric isomers having S,R,E configuration, R,S,E configuration, S,S,E configuration, and R,R,E configuration.
  • “Enantioselectivity value” or “E” refers to the ratio of specificity constants for each enantiomer of a compound undergoing chemical reaction or conversion and may be calculated (for the S-enantiomer) from the expression, E = K S / K SM K R / K RM = ln ⁢ ⌊ 1 - χ ⁡ ( 1 + ee p ) ⌋ ln ⁡ [ 1 - χ ⁡ ( 1 - ee p ) ] = ln ⁡ [ 1 - χ ⁡ ( 1 - ee S ) ] ln ⁡ [ 1 - χ ⁡ ( 1 + ee S ) ] ,
    where KS and KR are the 1st order rate constants for the conversion of the S- and R-enantiomers, respectively; KSM and KRM are the Michaelis constants for the S- and R-enantiomers, respectively; χ is the fractional conversion of the substrate; eep and ees are the enantiomeric excess of the product and substrate (reactant), respectively.
  • “Lipase Unit” or “LU” refers to the amount of enzyme (in g) that liberates 1 μmol of titratable butyric acid/min when contacted with tributyrin and an emulsifier (gum arabic) at 30° C. and pH 7.
  • “Solvate” refers to a molecular complex comprising a disclosed or claimed compound and a stoichiometric or non-stoichiometric amount of one or more solvent molecules (e.g., EtOH).
  • “Hydrate” refers to a solvate comprising a disclosed or claimed compound and a stoichiometric or non-stoichiometric amount of water.
  • “Pharmaceutically acceptable complexes, salts, solvates, or hydrates” refers to complexes, acid or base addition salts, solvates or hydrates of claimed and disclosed compounds, which are within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use.
  • “Pre-catalyst” or “catalyst precursor” refers to a compound or set of compounds that are converted into a catalyst prior to use.
  • “Treating” refers to reversing, alleviating, inhibiting the progress of, or preventing a disorder or condition to which such term applies, or to preventing one or more symptoms of such disorder or condition.
  • “Treatment” refers to the act of “treating,” as defined immediately above.
  • Table 1 lists abbreviation used throughout the specification.
    TABLE 1
    List of Abbreviations
    Abbreviation Description
    Ac Acetyl
    ACN acetonitrile
    AcNH acetylamino
    aq aqueous
    BES N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic
    acid
    BICINE N,N-bis(2-hydroxyethyl)glycine
    Bn benzyl
    Bu Butyl
    n-BuLi normal-butyl lithium
    Bu4NBr tetrabutylammonium bromide
    t-BuNH2 tertiary-butylamine
    t-BuOK potassium tertiary butyl oxide
    t-BuOMe tertiary butyl methyl ether
    t-BuONa sodium tertiary butyl oxide
    CBz benzyloxycarbonyl
    χ fractional conversion
    COD 1,5-cyclooctadiene
    DABCO 1,4-diazabicyclo[2.2.2]octane
    DBU 1,8-diazabicyclo[5.4.0]undec-7-ene
    DEAD diethylazodicarboxylate
    DIPEA diisopropylethylamine (Hunig's Base)
    DMAP 4-dimethylaminopyridine
    DMF dimethylformamide
    DMSO dimethylsulfoxide
    E Enantioselectivity value or ratio of specificity
    constants for each enantiomer of a compound
    undergoing chemical reaction or conversion
    ee (eep or ees) enantiomeric excess (of product or reactant)
    eq equivalents
    er enantiomeric ratio
    Et ethyl
    Et3N triethylamine
    Et2NH diethylamine
    EtOH ethyl alcohol
    EtOAc ethyl acetate
    h, min, s, d hours, minutes, seconds, days
    HEPES 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid
    HOAc acetic acid
    HPLC high performance liquid chromatography
    IAcOEt ethyl iodoacetate
    IPA isopropanol
    KS, KS 1st order rate constant for S- or R-enantiomer
    KSM, KRM Michaelis constant for S- or R-enantiomer
    LC/MS liquid chromatography mass spectrometry
    LDA Lithium diisopropylamide
    LiHMDS Lithium hexamethyldisilazide
    LTMP Lithium tetramethylpiperidide
    LU lipase unit
    Me methyl
    MeCl2 methylene chloride
    (R,R)-Me-DUPHOS (−)-1,2-bis((2R,5R)-2,5-
    dimethylphospholano)benzene
    MeI methyl iodide
    MeONa sodium methoxide
    MeOH methyl alcohol
    MES 2-morpholinoethanesulfonic acid
    MOPS 3-(N-morpholino)propanesulfonic acid
    Mpa mega Pascals
    Ms Mesyl or methylsulfonyl
    MTBE methyl tertiary butyl ether
    NMP N-methylpyrrolidone
    OTf− triflate (trifluoro-methanesulfonic acid anion)
    Ph phenyl
    Ph3P triphenylphosphine
    Ph3As triphenylarsine
    PIPES piperazine-1,4-bis(2-ethanesulfonic acid)
    RaNi Raney nickel
    RI refractive index
    RT room temperature (approximately 20° C.-25° C.)
    s/c substrate-to-catalyst molar ratio
    sp species
    TAPS N-[tris(hydroxymethyl)methyl]-3-
    aminopropanesulfonic acid
    TES N-[tris(hydroxymethyl)methyl]-2-
    aminoethanesulfonic acid
    Tf trifluoromethanesulfonyl (triflyl)
    TFA trifluoroacetic acid
    THF tetrahydrofuran
    TLC thin-layer chromatography
    TMEDA N,N,N′,N′-tetramethyl-1,2-ethylenediamine
    TRICINE N-[tris(hydroxymethyl)methyl]glycine
    Tris buffer tris(hydroxymethyl)aminomethane buffer
    TRITON B benzyltrimethylammonium hydroxide
    TRIZMA ® 2-amino-2-(hydroxymethyl)-1,3-propanediol
    Ts tosyl or p-toluenesulfonyl
    p-TSA para-toluene sulfonic acid
    v/v volume percent
    w/w weight (mass) percent
  • In some of the reaction schemes and examples below, certain compounds can be prepared using protecting groups, which prevent undesirable chemical reaction at otherwise reactive sites. Protecting groups may also be used to enhance solubility or otherwise modify physical properties of a compound. For a discussion of protecting group strategies, a description of materials and methods for installing and removing protecting groups, and a compilation of useful protecting groups for common functional groups, including amines, carboxylic acids, alcohols, ketones, aldehydes, and the like, see T. W. Greene and P. G. Wuts, Protecting Groups in Organic Chemistry (1999) and P. Kocienski, Protective Groups (2000), which are herein incorporated by reference in their entirety for all purposes.
  • In addition, some of the schemes and examples below may omit details of common reactions, including oxidations, reductions, and so on, which are known to persons of ordinary skill in the art of organic chemistry. The details of such reactions can be found in a number of treatises, including Richard Larock, Comprehensive Organic Transformations (1999), and the multi-volume series edited by Michael B. Smith and others, Compendium of Organic Synthetic Methods (1974-2003). Generally, starting materials and reagents may be obtained from commercial sources or may be prepared from literature sources.
  • Generally, the chemical transformations described throughout the specification may be carried out using substantially stoichiometric amounts of reactants, though certain reactions may benefit from using an excess of one or more of the reactants. Additionally, many of the reactions disclosed throughout the specification, including the enantioselective hydrolysis of the racemic diester (Formula 4) described in detail below, may be carried out at about RT, but particular reactions may require the use of higher or lower temperatures, depending on reaction kinetics, yields, and the like. Furthermore, many of the chemical transformations may employ one or more compatible solvents, which may influence the reaction rate and yield. Depending on the nature of the reactants, the one or more solvents may be polar protic solvents, polar aprotic solvents, non-polar solvents, or some combination. Any references in the disclosure to a concentration range, a temperature range, a pH range, a catalyst loading range, and so on, whether expressly using the word “range” or not, include the indicated endpoints.
  • The present invention provides materials and methods for preparing optically active γ-amino acids (Formula 1) including pharmaceutically acceptable salts, esters, amides, or prodrugs thereof. The compounds of Formula 1 include substituents R1 and R2, which are defined above. Useful compounds of Formula 1 thus include those in which R1 is a hydrogen atom and R2 is C1-12 alkyl, C3-12 cycloalkyl, or substituted C3-12 cycloalkyl, or those in which R2 is a hydrogen atom and R1 is C1-12 alkyl, C3-1 2 cycloalkyl, or substituted C3-12 cycloalkyl. Particularly useful compounds of Formula 1 include those in which R1 is a hydrogen atom and R2 is C1-6 alkyl or C3-7 cycloalkyl, or those in which R2 is a hydrogen atom and R1 is C1-6 alkyl or C3-7 cycloalkyl. Especially useful compounds of Formula 1 include those in which R1 is a hydrogen atom and R2 is C1-4 alkyl, such as pregabalin (Formula 9).
  • FIG. 1 shows a process for preparing optically active γ-amino acids (Formula 1). The process includes the step of contacting or combining a reaction mixture, which is comprised of a cyano-substituted diester (Formula 4) and water, with an enzyme to yield a product mixture that includes an optically active dicarboxylic acid monoester (Formula 3) and an optically-active diester (Formula 5). The cyano-substituted diester (Formula 4) has a stereogenic center, which is denoted by an asterisk (“*”), and as described below, may be prepared in accordance with a reaction scheme shown in FIG. 2. Prior to contacting the enzyme, the cyano-substituted diester (Formula 4) typically comprises a racemic (equimolar) mixture of the diester of Formula 5 and its opposite enantiomer. Substituents R1, R2, and R3 in Formula 3, Formula 4, and Formula 5, and substituent R4 in Formula 4 and Formula 5 are as defined above in connection with Formula 1. Generally, and unless stated differently, when a particular substituent identifier (R1, R2, R3, etc.) is defined for the first time in connection with a formula, the same substituent identifier used in a subsequent formula will have the same meaning as in the earlier formula.
  • The enzyme (or biocatalyst) may be any protein that, while having little or no effect on the compound of Formula 5, will catalyze the hydrolysis of its opposite enantiomer to yield the dicarboxylic acid monoester (Formula 3). Useful enzymes for enantioselectively hydrolyzing the compound of Formula 4 to Formula 3 may thus include hydrolases, including lipases, certain proteases, and other enantioselective esterases. Such enzymes may be obtained from a variety of natural sources, including animal organs and microorganisms. See, e.g., Table 2 for a non-limiting list of commercially available hydrolases.
    TABLE 2
    Commercially Available Hydrolases
    Enzyme Trade name
    Porcine Pancreatic Lipase Altus03
    CAL-A, lyophilized Altus11
    Candida lipolytica Lipase Altus12
    CAL-B, lyophilized Altus13
    Geotrichum candidum Lipase Altus28
    Pseudomonas aroginosa Lipase Altus50
    Aspergillus niger Lipase Amano Lipase A
    Pseudomonas cepacia Lipase Amano Lipase AH
    Pseudomonas fluorescens Lipase Amano Lipase AK
    Candida rugosa Lipase Amano Lipase AY
    Rhizopus delemar Lipase Amano Lipase D
    Rhizopus oryzae Lipase Amano Lipase F
    Penicillium camembertii Lipase Amano Lipase G
    Mucor javanicus Lipase Amano Lipase M
    Pseudomonas cepacia Lipase Amano Lipase PS
    Penicillium roqueforti Lipase Amano Lipase R
    Aspergillus sp. Protease BioCatalytics101
    Pseudomonas sp. Lipase BioCatalytics103
    Fungal Lipase BioCatalytics105
    Microbial, lyophilized Lipase BioCatalytics108
    CAL-B, lyophilized BioCatalytics110
    Candida sp., lyophilized BioCatalytics111
    CAL-A, lyophilized BioCatalytics112
    Thermomyces sp. Lipase BioCatalytics115
    Alcaligines sp., lyophilized Lipase BioCatalytics117
    Chromobacterium viscosum Lipase Altus 26
    CAL-B, L2 Sol Chriazyme L2 Sol
    Candida utilis Lipase Fluka6
    Rhizopus niveus Lipase Sigma L8
    Pseudomonas sp. Lipoprotein Lipase Sigma L13
    Thermomuces lanuginosus Lipase Sigma L9 Lipolase
    Thermomuces lanuginosus Lipase Sigma L10 Novo871
    Rhizomucor miehei Lipase Sigma L6 Palatase
    Pseudomonas species Lipase Sigma L14 Type XIII
    Wheat Germ Lipase Sigma L11
    Rhizopus arrhizus Lipase Sigma L7 Type XI
    Pancreatic Lipase 250 Valley Research V1
    Trypsin Protease Altus33
    Chymopapain Protease Altus38
    Bromelain Protease Altus40
    Aspergillus niger Protease Altus41
    Aspergillus oryzae Protease Altus42
    Penicillium sp. Protease Altus43
    Aspergillus sp. Protease Altus45
    Renin Calf Stomach Protease Sigma P24
    Subtilisin Carlsberg Protease Altus10
    Bacillus lentus Protease Altus53
    Aspergillus niger Protease Amano Acid Protease A
    Rhizopus niveus Protease Amano Acid Protease II
    Rhizopus niveus Protease Amano Newlase F
    Rhizopus oryzae Protease Amano Peptidase R
    Bacillus subtilis Protease Amano Proleather FGF
    Aspergillus oryzae Protease Amano Protease A
    Aspergillus oryzae Protease Amano Protease M
    Bacillus subtilis Protease Amano Protease N
    Aspergillus melleus Protease Amano Protease P
    Bacillus stearothermophilus Protease Amano Protease SG
    Pig Liver Esterase, lyophilized BioCat Chirazyme E1
    Pig Liver Esterase, lyophilized BioCat Chirazyme E2
    Streptomyces sp. Proteases BioCatalytics118
    Tritirachium album Protease Fluka P6 Proteinase K
    Bovine Pancreas Protease Sigma P18 alpha chymotrypsin I
    Streptomyces griseus Protease Sigma P16 Bacterial
    Bovine Pancreas Protease Sigma P21 Beta chymotrypsin
    Clostridium histolyticum Protease Sigma P13 Clostripain
    Bovine Intestine Protease Sigma P17 Enteropeptidase
    Porcine Intestine Protease Sigma P25 Enteropeptidase
    Bacillus sp. Protease Sigma P8 Esperase
    Aspergillus oryzae Protease Sigma P1 Flavourzyme
    Bacillus amyloliquefaciens Protease Sigma P5 Neutrase
    Carica papaya Protease Sigma P12 Papain
    Bacillus thermoproteolyticus rokko Sigma P10 Protease
    Pyrococcus furiosis Protease Sigma P14 Protease S
    Bacillus sp. Protease Sigma P9 Savinase
    Bovine Pancreas Protease Sigma P19 Type 1 (crude)
    Bacillus polymyxa Protease Sigma P7 Type IX
    Bacillus licheniformis Protease Sigma P6 Type VIII
    Aspergillus saitoi Protease Sigma P3 Type XIII
    Aspergillus sojae Protease Sigma P4 Type XIX
    Aspergillus oryzae Protease Sigma P2 Type XXIII
    Bacterial Protease Sigma P11 Type XXIV
    Rhizopus sp. Newlase Sigma15 Newlase
    Validase FP Conc. Valley05
    Bromelian Conc. Valley10
    Acylase from Aspergillus sp. Amano Am1
    Porcine kidney Acylase Sigma A-S2 Acylase I
    Penicillin G Acylase Altus06
    Esterase from Mucor meihei Fluka
    Candida rugosa Esterase Altus31
    Porcine Pancreatic Elastase Altus35
    Cholesterol Esterase BioCatalytics
    PLE - Ammonium Sulfate BioCatalytics 123
    Rabbit Liver Esterase Sigma ES2
    Cholesterol Esterase Pseudomonas sp. Sigma ES4
  • As shown in the Example section, useful enzymes for the enantioselective conversion of the cyano-substituted diester (Formula 4 and Formula 12) to the desired optically active dicarboxylic acid monoester (Formula 3 and Formula 11) include lipases. Particularly useful lipases include enzymes derived from the microorganism Thermomyces lanuginosus, such as those available from Novo-Nordisk A/S under the trade name LIPOLASE® (CAS no. 9001-62-1). LIPOLASE® enzymes are obtained by submerged fermentation of an Aspergillus oryzae microorganism genetically modified with DNA from Thermomyces lanuginosus DSM 4109 that encodes the amino acid sequence of the lipase. LIPOLASE® 100L and LIPOLASE® 100T are available as a liquid solution and a granular solid, respectively, each having a nominal activity of 100 kLU/g. Other forms of LIPOLASE® include LIPOLASE® 50L, which has half the activity of LIPOLASE® 100L, and LIPOZYME® 100L, which has the same activity of LIPOLASE® 100L, but is food grade.
  • Various screening techniques may be used to identify suitable enzymes. For example, large numbers of commercially available enzymes may be screened using high throughput screening techniques described in the Example section below. Other enzymes (or microbial sources of enzymes) may be screened using enrichment isolation techniques. Such techniques typically involve the use of carbon-limited or nitrogen-limited media supplemented with an enrichment substrate, which may be the racemic substrate (Formula 4) or a structurally similar compound. Potentially useful microorganisms are selected for further investigation based on their ability to grow in media containing the enrichment substrate. These microorganisms are subsequently evaluated for their ability to enantioselectively catalyze ester hydrolysis by contacting suspensions of the microbial cells with the racemic substrate and testing for the presence of the desired optically-active dicarboxylic acid monoester (Formula 3) using analytical methods such as chiral HPLC, gas-liquid chromatography, LC/MS, and the like.
  • Once a microorganism having the requisite hydrolytic activity has been isolated, enzyme engineering may be employed to improve the properties of the enzyme it produces. For example, and without limitation, enzyme engineering may be used to increase the yield and the enantioselectivity of the ester hydrolysis, to broaden the temperature and pH operating ranges of the enzyme, and to improve the enzyme's tolerance to organic solvents. Useful enzyme engineering techniques include rational design methods, such as site-directed mutagenesis, and in vitro-directed evolution techniques that utilize successive rounds of random mutagenesis, gene expression, and high throughput screening to optimize desired properties. See, e.g., K. M. Koeller & C.-H. Wong, “Enzymes for chemical synthesis,” Nature 409:232-240 (11 Jan. 2001), and references cited therein, the complete disclosures of which are herein incorporated by reference.
  • The enzyme may be in the form of whole microbial cells, permeabilized microbial cells, extracts of microbial cells, partially purified enzymes, purified enzymes, and the like. The enzyme may comprise a dispersion of particles having an average particle size, based on volume, of less than about 0.1 mm (fine dispersion) or of about 0.1 mm or greater (coarse dispersion). Coarse enzyme dispersions offer potential processing advantages over fine dispersions. For example, coarse enzyme particles may be used repeatedly in batch processes, or in semi-continuous or continuous processes, and may usually be separated (e.g., by filtration) from other components of the bioconversion more easily than fine dispersions of enzymes.
  • Useful coarse enzyme dispersions include cross-linked enzyme crystals (CLECs) and cross-linked enzyme aggregates (CLEAs), which are comprised primarily of the enzyme. Other coarse dispersions may include enzymes immobilized on or within an insoluble support. Useful solid supports include polymer matrices comprised of calcium alginate, polyacrylamide, EUPERGIT®, and other polymeric materials, as well as inorganic matrices, such as CELITE®. For a general description of CLECs and other enzyme immobilization techniques, see U.S. Pat. No. 5,618,710 to M. A. Navia & N. L. St. Clair. For a general discussion of CLEAs, including their preparation and use, see U.S. Patent Application No. 2003/0149172 to L. Cao & J. Elzinga et al. See also A. M. Anderson, Biocat. Biotransform, 16:181 (1998) and P. López-Serrano et al., Biotechnol. Lett. 24:1379-83 (2002) for a discussion of the application of CLEC and CLEA technology to a lipase. The complete disclosures of the abovementioned references are herein incorporated by reference for all purposes.
  • The reaction mixture may comprise a single phase or may comprise multiple phases (e.g., a two- or a three-phase system). Thus, for example, the enantioselective hydrolysis shown in FIG. 1 may take place in a single aqueous phase, which contains the enzyme, the initially racemic substrate (Formula 4), the undesired optically-active diester (Formula 5), and the desired optically-active dicarboxylic acid monoester (Formula 3). Alternatively, the reaction mixture may comprise a multi-phase system that includes an aqueous phase in contact with a solid phase (e.g., enzyme or product), an aqueous phase in contact with an organic phase, or an aqueous phase in contact with an organic phase and a solid phase. For example, the enantioselective hydrolysis may be carried out in a two-phase system comprised of a solid phase, which contains the enzyme, and an aqueous phase, which contains the initially racemic substrate, the undesired optically-active diester, and the desired optically-active dicarboxylic acid monoester.
  • Alternatively, the enantioselective hydrolysis may be carried out in a three-phase system comprised of a solid phase, which contains the enzyme, an organic phase that initially contains the racemic substrate (Formula 4), and an aqueous phase that initially contains a small fraction of the racemic substrate. Since the desired optically-active dicarboxylic acid monoester (Formula 3) has a lower pKa than the unreacted optically-active diester (Formula 5) and therefore exhibits greater aqueous solubility, the organic phase becomes enriched in the unreacted diester while the aqueous phase becomes enriched in the desired dicarboxylic acid monoester as the reaction proceeds.
  • The amounts of the racemic substrate (Formula 4) and the biocatalyst used in the enantioselective hydrolysis will depend on, among other things, the properties of the particular cyano-substituted diester and enzyme. Generally, however, the reaction may employ a substrate having an initial concentration of about 0.1 M to about 3.0 M, and in many cases, having an initial concentration of about 1.5 M to about 3.0 M. Additionally, the reaction may generally employ an enzyme loading of about 1% to about 10%, and in many cases, may employ an enzyme loading of about 3% to about 4% (v/v).
  • The enantioselective hydrolysis may be carried out over wide ranges of temperature and pH. For example, the reaction may be carried out at a temperature of about 10° C. to a temperature of about 50° C., but is typically carried out at about RT. Such temperatures generally permit substantially full conversion (e.g., about 42% to about 50%) of the racemate (Formula 4) in a reasonable amount of time (about 2 h to about 24 h) without deactivating the enzyme. Additionally, the enantioselective hydrolysis may be carried out at a pH of about 5 to a pH of about 10, more typically at a pH of about 6 to a pH of about 9, and often at a pH of about 6.5 to a pH of about 7.5.
  • In the absence of pH control, the reaction mixture pH will decrease as the hydrolysis of the substrate (Formula 4) proceeds because of the formation of the dicarboxylic acid monoester (Formula 3). To compensate for this change, the hydrolysis reaction may be run with internal pH control (i.e., in the presence of a suitable buffer) or may be run with external pH control through the addition of a base. Suitable buffers include potassium phosphate, sodium phosphate, sodium acetate, ammonium acetate, calcium acetate, BES, BICINE, HEPES, MES, MOPS, PIPES, TAPS, TES, TRICINE, Tris, TRIZMA®, or other buffers having a pKa of about 6 to a pKa of about 9. The buffer concentration generally ranges from about 5 mM to about 1 mM, and typically ranges from about 50 mM to about 200 mM. Suitable bases include aqueous solutions comprised of KOH, NaOH, NH4OH, etc., having concentrations ranging from about 0.5 M to about 15 M, or more typically, ranging from about 5 M to about 10 M. Other inorganic additives such as calcium acetate may also be used.
  • Following or during the enzymatic conversion of the racemate (Formula 4), the desired optically active dicarboxylic acid monoester (Formula 3) is isolated from the product mixture using standard techniques. For example, in the case of a single (aqueous) phase batch reaction, the product mixture may be extracted one or more times with a nonpolar organic solvent, such as hexane or heptane, which separates the desired dicarboxylic monoester (Formula 2) and the unreacted diester (Formula 5) in aqueous and organic phases, respectively. Alternatively, in the case of a multi-phase reaction employing aqueous and organic phases enriched in the desired monoester (Formula 3) and the unreacted diester (Formula 5), respectively, the monoester and diester may be separated batch-wise following reaction, or may be separated semi-continuously or continuously during the enantioselective hydrolysis.
  • As indicated in FIG. 1, the unreacted diester (Formula 5) may be isolated from the organic phase and racemized to yield the racemic substrate (Formula 4). The resulting racemate (Formula 4) may be recycled or combined with unconverted racemic substrate, which subsequently undergoes enzymatic conversion to Formula 3 as described above. Recycling the unreacted diester (Formula 5) increases the overall yield of the enantioselective hydrolysis above 50%, thereby increasing the atom economy of the method and lowering costs associated with disposal of the undesired enantiomers.
  • The treatment of the diester (Formula 5) with a base that is strong enough to abstract an acidic α-proton of the malonate moiety generally results in inversion of the stereogenic center and generation of the racemic substrate (Formula 4). Useful bases include organic bases, such as alkoxides (e.g., sodium ethoxide), linear aliphatic amines, and cyclic amines, and inorganic bases, such as KOH, NaOH, NH4OH, and the like. The reaction is carried out in a compatible solvent, including polar protic solvents, such as EtOH or aprotic polar solvents, such as MTBE. Reaction temperatures above RT typically improve the yield of the racemization process.
  • As shown in FIG. 1, the substantially enantiopure dicarboxylic acid monoester (Formula 3) may be converted to an optically active γ-amino acid (Formula 1) using at least three different methods. In one method, the monoester (Formula 3) is hydrolyzed in the presence of an acid catalyst or a base catalyst to yield an optically-active cyano-substituted dicarboxylic acid (Formula 6) or corresponding salt. The cyano moiety of the resulting dicarboxylic acid (or its salt) is reduced to yield an optically-active γ-amino dicarboxylic acid (Formula 7) or a corresponding salt, which is subsequently decarboxylated by treatment with an acid, by heating, or both, to yield the desired optically active γ-amino acid (Formula 1). The cyano moiety may be reduced via reaction with H2 in the presence of catalytic amounts of Raney nickel, palladium, platinum, and the like, or through reaction with a reducing agent, such as LiAlH4, BH3—Me2S, and the like. Useful acids for the hydrolysis and decarboxylation reactions include mineral acids, such as HClO4, H1, H2SO4, HBr, HCl, and the like. Useful base catalysts for the hydrolysis reaction include various alkali and alkaline earth metal hydroxides and oxides, including LiOH, NaOH, KOH, and the like.
  • In another method, the dicarboxylic acid monoester (Formula 3) undergoes reductive cyclization to form an optically-active cyclic 3-carboxy-pyrrolidin-2-one (Formula 2), which is subsequently treated with an acid to yield the desired enantiomerically-enriched γ-amino acid (Formula 1). The reductive cyclization may be carried out by reacting the monoester (Formula 3) with H2 in the presence of catalytic amounts of Raney nickel, palladium, platinum and the like. One or more acids may be used to hydrolyze and decarboxylate the resulting lactam acid (Formula 2), including mineral acids such as HClO4, H1, H2SO4, HBr, and HCl, and organic acids such as HOAc, TFA, p-TSA, and the like. The concentration of the acids may range from about 1N to about 12 N, and the amount of the acids may range from about 1 eq to about 7 eq. The hydrolysis and decarboxylation reactions may be carried out at a temperature of about RT or higher, or at a temperature of about 60° C. or higher, or at temperature in a range of about 60° C. to about 130° C.
  • In a third method, the ester moiety of the dicarboxylic acid monoester (Formula 3) is first hydrolyzed to give the cyano-substituted dicarboxylic acid (Formula 6 or its salt) as described above. The resulting dicarboxylic acid (or its salt) is subsequently decarboxylated to give an optically-active cyano-substituted carboxylic acid or its salt (Formula 8 in which R5 is a hydrogen atom, though R5 can also be C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl as noted below). The same conditions used to decarboxylate the lactam acid (Formula 2) or the γ-amino dicarboxylic acid (Formula 7) may be used. Instead of first hydrolyzing the ester moiety, the dicarboxylic acid monoester (Formula 3) may be first decarboxylated directly to a cyano substituted monoester (Formula 8) by heating the aqueous solution of the dicarboxylic acid monoester (as a salt) to a temperature of from about 50° C. to reflux. Krapcho conditions (DMSO/NaCl/water) may also be used. In either case, the cyano moiety of the compound of formula 8 is subsequently reduced to give the optically active γ-amino acid (Formula 1).
  • In addition to Raney nickel, a number of other catalysts may be used to reduce the cyano moiety of the compounds of Formula 3, 6 and 8. These include, without limitation, heterogeneous catalysts containing from about 0.1% to about 20%, and more typically, from about 1% to about 5%, by weight, of transition metals such as Ni, Pd, Pt, Rh, Re, Ru, and Ir, including oxides and combinations thereof, which are typically supported on various materials, including Al2O3, C, CaCO3, SrCO3, BaSO4, MgO, SiO2, TiO2, ZrO2, and the like. Many of these metals, including Pd, may be doped with an amine, sulfide, or a second metal, such as Pb, Cu, or Zn. Useful catalysts thus include palladium catalysts such as Pd/C, Pd/SrCO3, Pd/Al2O3, Pd/MgO, Pd/CaCO3, Pd/BaSO4, PdO, Pd black, PdCl2, and the like, containing from about 1% to about 5% Pd, based on weight. Other useful catalysts include Rh/C, Ru/C, Re/C, PtO2, Rh/C, RuO2, and the like.
  • The catalytic reduction of the cyano moiety is typically carried out in the presence of one or more polar solvents, including without limitation, water, alcohols, ethers, esters and acids, such as MeOH, EtOH, IPA, THF, EtOAc, and HOAc. The reaction may be carried out at temperatures ranging from about 5° C. to about 100° C., though reactions at RT are common. Generally, the substrate-to-catalyst ratio may range from about 1:1 to about 1000:1, based on weight, and H2 pressure may range from about atmospheric pressure, 0 psig, to about 1500 psig. More typically, the substrate-to-catalyst ratios range from about 4:1 to about 20: 1, and H2 pressures range from about 25 psig to about 150 psig.
  • All of the preceding methods may be used to convert the substantially enantiopure monoester (Formula 3) to the optically active γ-amino acid (Formula 1), but each may offer certain advantages over the others. For example, following acid workup of the process employing reductive cyclization, the lactam acid (Formula 2) may be isolated and purified by extracting it into an organic solvent, whereas the cyano-substituted dicarboxylic acid (Formula 6) may be more difficult to isolate because of its comparatively higher aqueous solubility. Isolation of the lactam acid (Formula 2) reduces the carryover of water-soluble impurities into the final product mixture and permits higher reactant concentration (e.g., about 1 M to about 2 M) during hydrolysis and decarboxylation, thereby increasing process throughput. Additionally, direct decarboxylation by heating the aqueous solution of the dicarboxylic acid monoester (Formula 3) affords the cyanomonoester (Formula 8) in high enantiomeric purity. This compound can be separated from the reaction medium by extraction in an organic solvent or by direct phase separation, ensuring efficient removal of inorganic impurities by the water phase. High reaction throughput and the avoidance of strongly acidic conditions are two advantages of this approach.
  • FIG. 2 illustrates a process for preparing cyano-substituted diesters (Formula 4), which may serve as substrates for the enzymatic enantioselective hydrolysis shown in FIG. 1. The process includes a crossed aldol condensation, which comprises reacting an unsymmetrical ketone or an aldehyde (Formula 17) with a malonic acid diester (Formula 18) in the presence of catalytic amounts of a base to yield an α,β-unsaturated malonic acid diester (Formula 19) in which R1, R2, R3, and R4 are as defined above in connection with Formula 1. This type of crossed aldol reaction is known as a Knoevenagel Condensation, which is described in a number of literature reviews. See, e.g., B. K. Wilk, Tetrahedron 53:7097-7100 (1997) and references cited therein, the complete disclosures of which are herein incorporated by reference for all purposes.
  • Generally, any base capable of generating an enolate ion from the malonic acid diester (Formula 18) may be used, including secondary amines, such as di-n-propylamine, di-1-propylamine, pyrrolidine, etc., and their salts. The reaction may include a carboxylic acid, such as HOAc, to neutralize the product and to minimize enolization of the unsymmetrical ketone or aldehyde (Formula 17). Reactions involving unsymmetrical ketones may also employ Lewis acids, such as titanium tetrachloride, zinc chloride, zinc acetate, and the like to facilitate reaction. The reaction is typically run in a hydrocarbon solvent under reflux conditions. Useful solvents include hexane, heptane, cyclohexane, toluene, methyl t-butyl ether, and the like, with azeotropic removal of water.
  • In a subsequent step, a cyanide source, such as HCN, acetone cyanohydrin, an alkali metal cyanide (NaCN, KCN, etc.), or an alkaline earth metal cyanide (magnesium cyanide, etc.), undergoes conjugate addition to the β-carbon of the α,β-unsaturated malonic acid diester (Formula 19). The reaction is typically carried out in one or more polar protic solvents, including EtOH, MeOH, n-propanol, isopropanol, or polar aprotic solvents, such as DMSO, and the like. Subsequent acid workup yields the cyano-substituted diester (Formula 4). For an application of the method depicted in FIG. 2 to prepare a pregabalin precursor (Formula 12), see U.S. Pat. No. 5,637,767 to Grote et al., which is herein incorporated by reference in its entirety and for all purposes.
  • The desired (S)- or (R)-enantiomers of any of the compounds disclosed herein may be further enriched through classical resolution, chiral chromatography, or recrystallization. For example, the optically active γ-amino acids (Formula 1 or Formula 9) may be reacted with an enantiomerically-pure compound (e.g., acid or base) to yield a pair of diastereoisomers, each composed of a single enantiomer, which are separated via, say, fractional recrystallization or chromatography. The desired enantiomer is subsequently regenerated from the appropriate diastereoisomer. Additionally, the desired enantiomer often may be further enriched by recrystallization in a suitable solvent when it is it available in sufficient quantity (e.g., typically not much less than about 85% ee, and in some cases, not much less than about 90% ee).
  • As described throughout the specification, many of the disclosed compounds have stereoisomers. Some of these compounds may exist as single enantiomers (enantiopure compounds) or mixtures of enantiomers (enriched and racemic samples), which depending on the relative excess of one enantiomer over another in a sample, may exhibit optical activity. Such stereoisomers, which are non-superimposable mirror images, possess a stereogenic axis or one or more stereogenic centers (i.e., chirality). Other disclosed compounds may be stereoisomers that are not mirror images. Such stereoisomers, which are known as diastereoisomers, may be chiral or achiral (contain no stereogenic centers). They include molecules containing an alkenyl or cyclic group, so that cis/trans (or Z/E) stereoisomers are possible, or molecules containing two or more stereogenic centers, in which inversion of a single stereogenic center generates a corresponding diastereoisomer. Unless stated or otherwise clear (e.g., through use of stereobonds, stereocenter descriptors, etc.) the scope of the present invention generally includes the reference compound and its stereoisomers, whether they are each pure (e.g., enantiopure) or mixtures (e.g., enantiomerically enriched or racemic).
  • Some of the compounds may also contain a keto or oxime group, so that tautomerism may occur. In such cases, the present invention generally includes tautomeric forms, whether they are each pure or mixtures.
  • Many of the compounds described in this disclosure, including those represented by Formula 1 and Formula 9, are capable of forming pharmaceutically acceptable salts. These salts include, without limitation, acid addition salts (including diacids) and base salts. Pharmaceutically acceptable acid addition salts include nontoxic salts derived from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, hydrofluoric, phosphorous, and the like, as well nontoxic salts derived from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Such salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate, and the like.
  • Pharmaceutically acceptable base salts include nontoxic salts derived from bases, including metal cations, such as an alkali or alkaline earth metal cation, as well as amines. Examples of suitable metal cations include, without limitation, sodium cations (Na+), potassium cations (K+), magnesium cations (Mg2+), calcium cations (Ca2+), and the like. Examples of suitable amines include, without limitation, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, procaine, and t-butyl amine. For a discussion of useful acid addition and base salts, see S. M. Berge et al., “Pharmaceutical Salts,” 66 J. of Pharm. Sci., 1-19 (1977); see also Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (2002).
  • One may prepare a pharmaceutically acceptable acid addition salt (or base salt) by contacting a compound's free base (or free acid) or zwitterion with a sufficient amount of a desired acid (or base) to produce a nontoxic salt. If the salt precipitates from solution, it may be isolated by filtration; otherwise, the salt may be recovered by evaporating the solvent. One may also regenerate the free base (or free acid) by contacting the acid addition salt with a base (or the base salt with an acid). Though certain physical properties of the free base (or free acid) and its respective acid addition salt (or base salt) may differ (e.g., solubility, crystal structure, hygroscopicity, etc.), a compound's free base and acid addition salt (or its free acid and base salt) are otherwise the same for purposes of this disclosure.
  • Disclosed and claimed compounds may exist in both unsolvated and solvated forms and as other types of complexes besides salts. Useful complexes include clathrates or compound-host inclusion complexes where the compound and host are present in stoichiometric or non-stoichiometric amounts. Useful complexes may also contain two or more organic, inorganic, or organic and inorganic components in stoichiometric or non-stoichiometric amounts. The resulting complexes may be ionized, partially ionized, or non-ionized. For a review of such complexes, see J. K. Haleblian, J. Pharm. Sci. 64(8):1269-88 (1975). Pharmaceutically acceptable solvates also include hydrates and solvates in which the crystallization solvent may be isotopically substituted, e.g. D2O, d6-acetone, d6-DMSO, etc. Generally, for the purposes of this disclosure, references to an unsolvated form of a compound also include the corresponding solvated or hydrated form of the compound.
  • The disclosed compounds also include all pharmaceutically acceptable isotopic variations, in which at least one atom is replaced by an atom having the same atomic number, but an atomic mass different from the atomic mass usually found in nature. Examples of isotopes suitable for inclusion in the disclosed compounds include, without limitation, isotopes of hydrogen, such as 2H and 3H; isotopes of carbon, such as 13C and 14C; isotopes of nitrogen, such as 15N; isotopes of oxygen, such as 17O and 18O; isotopes of phosphorus, such as 31P and 32P; isotopes of sulfur, such as 35S; isotopes of fluorine, such as 18F; and isotopes of chlorine, such as 36Cl. Use of isotopic variations (e.g., deuterium, 2H) may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements. Additionally, certain isotopic variations of the disclosed compounds may incorporate a radioactive isotope (e.g., tritium, 3H, or 14C), which may be useful in drug and/or substrate tissue distribution studies.
    • EXAMPLES
  • The following examples are intended to be illustrative and non-limiting, and represent specific embodiments of the present invention.
  • General Materials and Methods
  • Enzyme screening was carried out using a 96-well plate, which is described in D. Yazbeck et al., Synth. Catal. 345:524-32 (2003), the complete disclosure of which is herein incorporated by reference for all purposes. All enzymes used in the screening plate (see Table 2) were obtained from commercial enzyme suppliers including Amano (Nagoya, Japan), Roche (Basel, Switzerland), Novo Nordisk (Bagsvaerd, Denmark), Altus Biologics Inc. (Cambridge, Mass.), Biocatalytics (Pasadena, Calif.), Toyobo (Osaka, Japan), Sigma-Aldrich (St. Louis, Mo.) and Fluka (Buchs, Switzerland). The screening reactions were performed in an Eppendorf Thermomixer-R (VWR). Subsequent larger scale enzymatic resolutions employed LIPOLASE® 100L and LIPOLASE® 100T, which are available form Novo-Nordisk A/S (CAS no. 9001-62-1).
  • Nuclear Magnetic Resonance
  • Three hundred MHz 1H NMR and 75 MHz 13C NMR spectra were obtained on a BRUKER 300 UltraShield™ equipped with a 5 mm auto switchable PHQNP probe. Spectra were generally acquired near RT, and standard autolock, autoshim and autogain routines were employed. Samples were usually spun at 20 Hz for 1D experiments. 1H NMR spectra were acquired using 30-degree tip angle pulses, 1.0 s recycle delay, and 16 scans at a resolution of 0.25 Hz/point. The acquisition window was typically 8000 Hz from +18 to −2 ppm (Reference TMS at 0 ppm) and processing was with 0.3 Hz line broadening. Typical acquisition time was 5-10 s. Regular 13C NMR spectra were acquired using 30-degree tip angle pulses, 2.0 s recycle delay, and 2048 scans at a resolution of 1 Hz/point. Spectral width was typically 25 KHz from +235 to −15 ppm (Reference TMS at 0 ppm). Proton decoupling was applied continuously and 1 Hz line broadening was applied during processing. Typical acquisition time was 102 min.
  • Mass Spectrometry
  • Mass Spectrometry was performed on a HEWLETT PACKARD 1100MSD using HP Chemstation Plus Software. The LC was equipped with an Agilent 1100 quaternary LC system and an Agilent liquid handler as an autosampler. Data were acquired under electron spray ionization with ACN/water (containing 0.1% formic acid) as the solvent (10% ACN to 90%, 7 min). Temperatures: probe was 350° C., source was 150° C. Corona discharge was 3000 V for positive ion and 3000 V for negative ion.
  • High Performance Liquid Chromatography
  • High Performance Liquid Chromatography (HPLC) was performed on a series 1100 AGILENT TECHNOLOGIES instrument equipped with an Agilent 220 HPLC auto sampler, quaternary pump, and a UV detector. The LC was PC controlled using HP Chemstation Plus Software. Normal Phase chiral HPLC was performed using Chiral HPLC columns obtained from Chiral Technologies (Exton, Pa.) and Phenomenex (Torrance, Calif.).
  • Gas Chromatography
  • Gas Chromatography (GC) was performed on a 110 volt Agilent 6890N network GC system equipped with an FID detector with electrometer, a 7683 Series split/splitless capillary injector, a relay board that monitors four external events, and an inboard printer/plotter. Enantiomeric excess of the diester (Formula 13, R3=R4=Et) and monoester (Formula 11, R3=Et) were performed using a CHIRALDEX G-TA column (30 m×0.25 mm), with helium carrier gas, and at 135° C. Under such conditions, the monoester decomposed to give (S)-3-cyano-5-methyl-hexanoic acid ethyl ester, and ee was determined based on the decomposition product. The chiral GC columns used in analysis were obtained from Astec, Inc (Whippany, N.J.).
    • Example 1 Enzyme screening via enzymatic hydrolysis of (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) to yield (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid (Formula 21)
  • Figure US20050283023A1-20051222-C00019
  • Enzyme screening was carried out using a screening kit comprised of individual enzymes deposited in separate wells of a 96-well plate, which was prepared in advance in accordance with a method described in D. Yazbeck et al., Synth. Catal. 345:524-32 (2003). Each of the wells had an empty volume of 0.3 ml (shallow well plate). One well of the 96-well plate contained only phosphate buffer (10 μL, 0.1 M, pH 7.2), another well contained only ACN (10 μL), and each of the remaining wells contained one of the 94 enzymes listed in Table 2 (10 μL, 100 mg/mL). Prior to use, the screening kit was removed from storage at −80° C. and the enzymes were allowed to thaw at RT for about 5 min. Potassium phosphate buffer (85 μL, 0.1 M, pH 7.2) was dispensed into each of the wells using a multi-channel pipette. Concentrated substrate (Formula 20, 5 μL) was subsequently added to each well via a multi-channel pipette and the 96 reaction mixtures were incubated at 30° C. and 750 rpm. The reactions were quenched and sampled after 24 h by transferring each of the reaction mixtures into separate wells of a second 96-well plate. Each of the wells had an empty volume of 2 mL (deep well plate) and contained EtOAc (1 mL) and HCl (1N, 100 μL). The components of each well were mixed by aspirating the well contents with a pipette. The second plate was centrifuged and 100 μL of the organic supernatant was transferred from each well into separate wells of a third 96-well plate (shallow plate). The wells of the third plate were subsequently sealed using a penetrable mat cover. Once the wells were sealed, the third plate was transferred to a GC system for determination of optical purity (ee).
  • Table 3 lists enzyme, trade name, supplier, and E value for some of the enzymes that were screened. For a given enzyme, the E value may be interpreted as the relative reactivity of a pair of enantiomers (substrates). The E values listed in Table 3 were calculated from HPLC data (fractional conversion, χ, and ee) using a computer program called Ee2, which is available from the University of Graz. Generally, enzymes exhibiting S-selectivity and an E value of about 35 or greater are suitable for scale-up.
    TABLE 3
    Results from Screening Reactions of Example 1
    Enzyme Trade name Supplier E Value
    S-Selective
    Thermomyces lanuginosus Lipolase Novozymes >200
    Lipase
    Rhizopus delemar Lipase Lipase D Amano >200
    Rhizopus niveus Lipase L-9406 Sigma 66
    Rhizomucor miehei Esterase 46059 Fluka 52
    Pseudomonas sp. Lipase  103 Biocatalytics 51
    Rhizomucor miehei Lipase Palatase 20000 Novozymes 41
    Rhizopus oryzae Lipase FAP15 Amano 35
    Candida antarctica Lipase - A CAL-A Novozymes 5
    Candida antarctica Lipase - B CAL-B, Novozymes 3
    Chirazyme L-2
    Marginally S-Selective
    Pig liver Esterase PLE-AS Biocatalytics <2
    Enteropeptidase Sigma <2
    Porcine kidney Acylase Sigma <2
    Cholesterol Esterase Biocatalytics <2
    R-Selective
    Streptomyces griseus Protease Sigma 20
    Streptomyces sp. Protease  118 Biocatalytics 11
    • Example 2 Enzymatic resolution of (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) to yield (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23) and (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22)
  • Figure US20050283023A1-20051222-C00020
  • A reactor (392 L) equipped with overhead stirring was charged with potassium phosphate buffer (292.2 L, 10 mM, pH 8.0) and LIPOLASE® 100L, type EX (3.9 L). The mixture was stirred at 800 RPM for 1 min and KOH (2 M) was added to adjust the pH to 8.0. (R/S)-3-Cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20, 100 kg) was added, and the resulting mixture was titrated with NaOH aq (50%) during hydrolysis to maintain a pH of 8.0. The extent of reaction was monitored by HPLC (C18 column, 4.6 mm×150 mm, detection at 200 nm). Upon reaching a conversion of about 40-45% (e.g., after about 24 h) the reaction mixture was transferred to a separatory funnel. The aqueous mixture was extracted with heptane (205 L). EtOH (absolute) was added (up to about 5% v/v) to disrupt a light emulsion that formed, and the aqueous and organic layers were separated. The extraction step was repeated twice, and the aqueous layer containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23) may be further concentrated under vacuum (e.g., 25-50% of its original volume). The organic layers containing (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22) were combined, dried, and concentrated. The resulting diethyl ester was subsequently racemized in accordance with Example 6. MS m/z [+H]+227. 1H NMR (300 MHz, D2O): δ 2.35 (dd, 6H), 2.70 (t, 3H), 2.85 (m, 1H), 2.99 (m, 1H), 3.25 (m, 1H), 4.75 (m, 1H), 5.60 (q, 2H). 13C NMR (75 ppm, D2O) δ 172.19, 171.48, 122.85, 62.70, 59.49, 40.59, 31.83, 27.91, 23.94, 21.74, 14.77.
    • Example 3 Enzymatic resolution of (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) to yield (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23) and (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22)
  • A reactor (3.92 L) equipped with overhead stirring is charged with calcium acetate buffer (1.47 L, 100 mM, pH 7.0) and (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20, 1 kg). The mixture is stirred at 1100 RPM for 5 min and KOH (5 M) is added to adjust the pH to 7.0. LIPOLASE® 100L, type EX (75 mL) is added and the resulting mixture is titrated with KOH (5 M) during hydrolysis to maintain a pH of 7.0. The extent of reaction is monitored by HPLC (C18 column, 4.6 mm×150 mm, detection at 200 nm). Upon reaching a conversion of about 42% to 45% (e.g., after about 20-25 h) the reaction mixture is transferred to a separatory funnel. The aqueous mixture is extracted with hexane (100% v/v). EtOH (absolute) is added (up to about 5% v/v) to disrupt a light emulsion that forms, and the aqueous and organic layers are separated. The extraction step is repeated twice to obtain an aqueous layer containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23), which may be used in subsequent transformations without isolation. The organic layers containing (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22) are combined, dried, and concentrated. The resulting diethyl ester is subsequently racemized in accordance with Example 6.
    • Example 4 Preparation of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10) from (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23)
  • Figure US20050283023A1-20051222-C00021
  • A vessel was charged with an aqueous solution containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid potassium salt (Formula 23, 411 L from Example 2). Raney Nickel (50% aq solution, Sigma-Aldrich) was added to the mixture, and hydrogen gas was introduced into the vessel over a 20 h period to maintain a pressure of 50 psig in the vessel headspace throughout reaction. The hydrogenation reaction was monitored by H2 uptake and HPLC analysis (C18 column, 4.6 mm×150 mm, detection at 200 nm) of the vessel contents. Following reaction, the aqueous mixture was filtered to remove the Raney Ni catalyst. The pH of the concentrated solution was adjusted to 3.0 using 37% HCl (about 14 L). The resulting solution was extracted three times with EtOAc (50% v/v). The combined organic layers were concentrated under vacuum to afford (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10). MS m/z [M+H]+186.1130. 13C NMR (75 ppm, CDCl3) δ 175.67, 172.23, 54.09, 47.62, 43.69, 37.22, 26.31, 23.34, 22.54. Yield 40-42%; 97% ee.
    • Example 5 Preparation of pregabalin (Formula 9) from (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10)
  • Figure US20050283023A1-20051222-C00022
  • A reactor vessel (60 L) was charged with (S)4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10), HCl (36-38%, 30 L), and water (29 L). HOAc (1 L) was added to the solution and the resulting slurry was heated for 36-38 h at 80° C. and for an additional 6 h at 110° C. The extent of reaction was monitored by HPLC (C18 column, 4.6 mm×150 mm, detection at 200 nm). Water and excess HCl were evaporated to afford an oil, which was washed with MTBE (2×15 L). Water was added to the oil and the mixture was stirred until the solution cleared. The pH of the solution was adjusted to 5.2-5.5 using KOH (about 6 kg), which resulted in the precipitation of pregabalin. The mixture was heated to 80° C. and subsequently cooled to 4° C. After 10 h, crystalline pregabalin was filtered and washed with IPA (12 L). The filtrate was concentrated under vacuum to afford a residual oil. Water (7.5 L) and EtOH (5.0 L) were added to the residual oil and the resulting mixture was heated to 80° C. and then cooled to 4° C. After 10 h, a second crop of pregabalin crystals were filtered and washed with IPA (1 L). The combined pregabalin crystals were dried in a vacuum oven at 45° C. for 24 h. MS m/z [M+H]+160.1340. 1H NMR (300 MHz, D2O): δ 2.97 (dd, J=5.4, 12.9 Hz, 1H), 2.89 (dd, J=6.6, 12.9 Hz, 1H), 2.05-2.34 (m, 2H), 1.50-1.70 (sept, J=6.9 Hz, 1H), 1.17 (t, J=7.0 Hz, 2H), 0.85 (dd, J=2.2, 6.6 Hz, 6H). 13C NMR (75 ppm, D2O) δ 181.54, 44.32, 41.28, 32.20, 24.94, 22.55, 22.09. Yield 80-85%; ee>99.5%.
    • Example 6 Preparation of (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) via racemization of (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22)
  • Figure US20050283023A1-20051222-C00023
  • A reactor was charged with (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22, 49.5 kg) and EtOH (250 L). Sodium ethoxide (21% w/w in EtOH, 79.0 L, 1.1 eq) was added to the mixture, which was heated to 80° C. for 20 h. After completion of the reaction, the mixture was allowed to cool to RT and was neutralized by adding HOAc (12.2 L). Following evaporation of the EtOH, MTBE (150 L) was added to the mixture, and the resulting solution was filtered and evaporated to afford (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) in quantitative yield.
    • Example 7 Preparation (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24) from (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid (Formula 21)
  • Figure US20050283023A1-20051222-C00024
  • A 50 mL round bottomed flask was charged with (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid (Formula 21, 3.138 g, 13.79 mmol), NaCl (927 mg, 1.15 eq), de-ionized water (477 μL, 1.92 eq) and DMSO (9.5 mL). The resulting mixture was heated to 88° C. and maintained at that temperature for 17 h. A sample was taken for LC and LC/MS analyses, which showed the presence of the starting material (Formula 21) and the products (Formula 24 and Formula 25). The temperature of the mixture was subsequently increased to 135° C. and allowed to react for an additional 3.5 h. A second sample was taken for LC and LC/MS analysis, which showed the absence of starting material (Formula 21) and showed, in addition to the desired products (Formula 24 and Formula 25), the presence of unidentified byproducts. (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24): 97.4% ee after 88° C.; 97.5% ee after 135° C.
    • Example 8 Determination of the optical purity (ee) of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10)
  • The optical purity of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid (Formula 10) was determined via a derivatization method. A sample of (S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid was esterified with EtOH in the presence of a catalytic amount of dry HCl in dioxane at 70° C. The resulting lactam ester was analyzed by HPLC (CHIRALPAK AD-H, 4.6 mm×250 mm) using a mobile phase of hexane and EtOH (95:5), a flow rate of 1.0 mL/min, injection volume of 10 μL, column temperature of 35° C., and detection at 200 nm.
    • Example 9 Determination of the optical purity (ee) of pregabalin (Formula 9)
  • The optical purity of pregabalin was analyzed via a derivatization method. A sample of pregabalin was derivatized with Marfey's reagent (1-fluoro-24-dinitrophenyl-5-L-alanine amide) and then analyzed by HPLC (LUNA C18(2) column, 0.46 mm×150 mm, 3 μm) using a mobile phase of aqueous NaPO4 (20 nM, pH 2.0) and ACN (90:10 for 10 min, 10:90 for 3 min, 90:10 for 5 min), a flow rate of 1.2 mL/min, an injection volume of 10 μL, column temperature of 35° C., and detection at 200 nm.
    • Example 10 Enzymatic resolution of (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20) to yield (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid sodium salt (Formula 23) and (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22)
  • Figure US20050283023A1-20051222-C00025
  • A reactor (16000 L) equipped with overhead stirring is charged with calcium acetate (254 kg), deionized water (1892.7 kg) and LIPOZYME® TL 100 L (food grade LIPOLASE®, 983.7 kg). After complete mixing, (R/S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 20, 9000 kg, 85% purity assay) is charged and the mixture is stirred for 24 h. NaOH (2068 kg of a 30% solution) is added over the course of the reaction to maintain the pH at 7.0. The extent of reaction is monitored by HPLC (C18 column, 4.6 mm×150 mm, detection at 200 nm). Upon reaching a conversion of about 42% to 45% (e.g., after about 20-25 h) the titrator and stirring are stopped. The organic phase is immediately separated and the aqueous phase is washed twice with toluene (780 kg). The aqueous layer containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid sodium salt (Formula 23) is used in subsequent transformations (Example 11) without isolation. The organic layers containing (R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester (Formula 22) are combined and concentrated. The resulting diethyl ester is subsequently racemized in accordance with Example 6.
    • Example 11 Preparation (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24) from (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid sodium salt (Formula 23)
  • Figure US20050283023A1-20051222-C00026
  • A reactor (16000 L) equipped with overhead stirring is charged with the final aqueous solution from Example 10 (9698.6 L, containing (3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid sodium salt, Formula 23), NaCl (630 kg) and toluene (900 L). The mixture is stirred for 2 h under refluxing conditions (75-85° C.). The stirring is stopped; the organic phase is immediately separated and the aqueous phase is washed twice with toluene (900 L). The organic layers, which contain (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24) are combined and concentrated. The ethyl ester (Formula 24) is subsequently hydrolyzed in accordance with Example 12.
    • Example 12 Preparation of (S)-3-cyano-5-methyl-hexanoic acid potassium salt (Formula 26) from (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24)
  • Figure US20050283023A1-20051222-C00027
  • A reactor (12000 L) equipped with overhead stirring is charged with (S)-3-cyano-5-methyl-hexanoic acid ethyl ester (Formula 24, 2196 L from Example 11). KOH (1795.2 kg, 45% solution, w/w) and H2O (693.9 kg) are added to the reaction mixture with vigorous stirring. The temperature is maintained at 25° C. After 4 h, the reaction mixture is charged to a hydrogenation vessel (Example 13) with no further work-up.
    • Example 13 Preparation of pregabalin (Formula 9) from (S)-3-cyano-5-methyl-hexanoic acid potassium salt (Formula 26)
  • Figure US20050283023A1-20051222-C00028
  • A hydrogenator (12000 L) is charged with water (942.1 L) and with the reaction mixture from Example 12, which contains (S)-3-cyano-5-methyl-hexanoic acid potassium salt (Formula 26,4122.9 L). A Raney nickel suspension (219.6 kg, 50% w/w in H2O) is added. The hydrogenation is conducted under 50 psig at 35° C. After 6 h, the Raney nickel is filtered and the resulting filtrate is transferred to a reactor (16000 L) for crystallization. After adding H2O (1098 L), the pH of the solution is adjusted to 7.0-7.5 using HOAc (864.7 kg). The resulting precipitate is filtered and washed once with H2O (549 L) and twice with IPA (2,586 L each). The solid is recrystallized with IPA (12296 L) and H2O (6148 L). The mixture is heated to 70° C. and subsequently cooled to 4° C. After 5-10 h, the crystalline solid is filtered, washed with IPA (5724 L), and dried in a vacuum oven at 45° C. for 24 h to give pregabalin as a white crystalline solid (1431 kg, 30.0% overall yield, 99.5% purity and 99.75% ee).
  • It should be noted that, as used in this specification and the appended claims, singular articles such as “a,” “an,” and “the,” may refer to a single object or to a plurality of objects unless the context clearly indicates otherwise. Thus, for example, reference to a composition containing “a compound” may include a single compound or two or more compounds. It is to be understood that the above description is intended to be illustrative and not restrictive. Many embodiments will be apparent to those of skill in the art upon reading the above description. Therefore, the scope of the invention should be determined with references to the appended claims and includes the full scope of equivalents to which such claims are entitled. The disclosures of all articles and references, including patents, patent applications and publications, are herein incorporated by reference in their entirety and for all purposes.

Claims (15)

1. A method of making a compound of Formula 1,
Figure US20050283023A1-20051222-C00029
or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, in which
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl,
the method comprising:
(a) reacting a compound of Formula 2,
Figure US20050283023A1-20051222-C00030
or a salt thereof, with an acid and water to yield the compound of Formula 1 or a salt thereof; and
(b) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 2 are as defined in Formula 1.
2. The method of claim 1, wherein R1 is a hydrogen atom and R is isobutyl.
3. The method of claim 1, further comprising reducing a cyano moiety of a compound of Formula 3,
Figure US20050283023A1-20051222-C00031
or a salt thereof, to yield the compound of Formula 2 or a salt thereof, wherein
R1 and R2 in Formula 3 are as defined in Formula 1; and
R3 in Formula 3 is C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
4. The method of claim 3, wherein R1 is a hydrogen atom and R2 is isobutyl.
5. The method of claim 3, further comprising:
(a) contacting a compound of Formula 4,
Figure US20050283023A1-20051222-C00032
with an enzyme to yield the compound of Formula 3, or a salt thereof, and a compound of Formula 5,
Figure US20050283023A1-20051222-C00033
wherein the enzyme is adapted to enantioselectively hydrolyze the compound of Formula 4 to the compound of Formula 3 or a salt thereof;
(b) isolating the compound of Formula 3 or a salt thereof; and
(c) optionally racemizing the compound of Formula 5 to yield the compound of Formula 4, wherein
R1, R2, and R3 in Formula 4 and Formula 5 are as defined in Formula 3; and
R4 in Formula 4 and Formula 5 is the same as or different than R3 and is C1-2 alkyl, C3-12 cycloalkyl, or aryl-C3-12 alkyl.
6. The method of claim 5, wherein R1 is a hydrogen atom and R is isobutyl.
7. A method of making a compound of Formula 1,
Figure US20050283023A1-20051222-C00034
or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, in which
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl,
the method comprising:
(a) reducing a cyano moiety of a compound of Formula 6,
Figure US20050283023A1-20051222-C00035
or a salt thereof, to yield a compound of Formula 7,
Figure US20050283023A1-20051222-C00036
or a salt thereof;
(b) decarboxylating the compound of Formula 7 or a salt thereof to yield the compound of Formula 1 or a salt thereof; and
(c) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 6 and in Formula 7 are as defined in Formula 1.
8. The method of claim 7, wherein R1 is a hydrogen atom and R2 is isobutyl.
9. A method of making a compound of Formula 1,
Figure US20050283023A1-20051222-C00037
or a pharmaceutically acceptable complex, salt, solvate or hydrate thereof, in which
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl,
the method comprising:
(a) reducing a cyano moiety of a compound of Formula 8,
Figure US20050283023A1-20051222-C00038
or a salt thereof, to yield the compound of Formula 1 or a salt thereof; and
(b) optionally converting the compound of Formula 1 or a salt thereof into a pharmaceutically acceptable complex, salt, solvate or hydrate, wherein R1 and R2 in Formula 8 are as defined in Formula 1, and R5 in Formula 8 is hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
10. The method of claim 9, wherein R1 is a hydrogen atom and R2 is isobutyl.
11. A method of making a compound of Formula 3,
Figure US20050283023A1-20051222-C00039
or a salt thereof, in which
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl, and
R3 is C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl,
the method comprising:
(a) contacting a compound of Formula 4,
Figure US20050283023A1-20051222-C00040
with an enzyme to yield the compound of Formula 3 and a compound of Formula 5,
Figure US20050283023A1-20051222-C00041
wherein the enzyme is adapted to enantioselectively hydrolyze the compound of Formula 4 to the compound of Formula 3 or a salt thereof;
(b) isolating the compound of Formula 3 or a salt thereof; and
(c) optionally racemizing the compound of Formula 5 to yield the compound of Formula 4, wherein
R1, R2, and R3 in Formula 4 and Formula 5 are as defined above in Formula 3; and
R4 in Formula 4 and Formula 5 is the same as or different than R3 and is C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl.
12. The method of claim 11, wherein R1 is a hydrogen atom and R is isobutyl.
13. A compound of Formula 2,
Figure US20050283023A1-20051222-C00042
including salts thereof, wherein
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl, provided that when one of the substituents represented by R1 or R2 is hydrogen, the other substituent is not C1-3 alkyl or C5 alkyl.
14. A compound of Formula 27,
Figure US20050283023A1-20051222-C00043
including salts thereof, wherein
R1 and R2 are different and are each independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, and substituted C3-12 cycloalkyl, provided that when one of the substituents represented by R1 or R2 is a hydrogen atom, the other substituent is not methyl; and
R5 and R6 are independently selected from hydrogen atom, C1-12 alkyl, C3-12 cycloalkyl, or aryl-C1-6 alkyl, provided that R5 and R6 are different if not both hydrogen atoms.
15. A compound selected from:
3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
(3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
(2S,3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
(2R,3S)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid,
3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester,
(R)-3-cyano-2-ethoxycarbonyl-5-methyl-hexanoic acid ethyl ester,
4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid,
(S)-4-isobutyl-2-oxo-pyrrolidine-3-carboxylic acid,
3-cyano-2-carboxy-5-methyl-hexanoic acid,
(S)-3-cyano-2-carboxy-5-methyl-hexanoic acid,
3-aminomethyl-2-carboxy-5-methyl-hexanoic acid, and
(S)-3-aminomethyl-2-carboxy-5-methyl-hexanoic acid,
including salts thereof and opposite enantiomers thereof.
US11/157,585 2004-06-21 2005-06-21 Preparation of pregabalin and related compounds Abandoned US20050283023A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
US11/157,585 US20050283023A1 (en) 2004-06-21 2005-06-21 Preparation of pregabalin and related compounds
US12/254,336 US7838686B2 (en) 2004-06-21 2008-10-20 Preparation of pregabalin and related compounds
US12/951,429 US8044227B2 (en) 2004-06-21 2010-11-22 Preparation of pregabalin and related compounds
US13/247,032 US8134023B2 (en) 2004-06-21 2011-09-28 Preparation of pregabalin and related compounds

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US58167104P 2004-06-21 2004-06-21
US62903404P 2004-11-18 2004-11-18
US11/157,585 US20050283023A1 (en) 2004-06-21 2005-06-21 Preparation of pregabalin and related compounds

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US12/254,336 Continuation US7838686B2 (en) 2004-06-21 2008-10-20 Preparation of pregabalin and related compounds

Publications (1)

Publication Number Publication Date
US20050283023A1 true US20050283023A1 (en) 2005-12-22

Family

ID=35295367

Family Applications (4)

Application Number Title Priority Date Filing Date
US11/157,585 Abandoned US20050283023A1 (en) 2004-06-21 2005-06-21 Preparation of pregabalin and related compounds
US12/254,336 Expired - Lifetime US7838686B2 (en) 2004-06-21 2008-10-20 Preparation of pregabalin and related compounds
US12/951,429 Active US8044227B2 (en) 2004-06-21 2010-11-22 Preparation of pregabalin and related compounds
US13/247,032 Expired - Lifetime US8134023B2 (en) 2004-06-21 2011-09-28 Preparation of pregabalin and related compounds

Family Applications After (3)

Application Number Title Priority Date Filing Date
US12/254,336 Expired - Lifetime US7838686B2 (en) 2004-06-21 2008-10-20 Preparation of pregabalin and related compounds
US12/951,429 Active US8044227B2 (en) 2004-06-21 2010-11-22 Preparation of pregabalin and related compounds
US13/247,032 Expired - Lifetime US8134023B2 (en) 2004-06-21 2011-09-28 Preparation of pregabalin and related compounds

Country Status (32)

Country Link
US (4) US20050283023A1 (en)
EP (1) EP1831154B1 (en)
JP (5) JP4174551B2 (en)
KR (3) KR100847928B1 (en)
CN (2) CN102102114B (en)
AP (1) AP2466A (en)
AR (1) AR049446A1 (en)
AT (1) ATE455093T1 (en)
AU (1) AU2005256945B2 (en)
BR (1) BRPI0512347B8 (en)
CA (1) CA2571040C (en)
CR (2) CR8765A (en)
CY (1) CY1110337T1 (en)
DE (1) DE602005018965D1 (en)
DK (1) DK1831154T3 (en)
EA (3) EA015418B1 (en)
ES (1) ES2336014T3 (en)
GE (1) GEP20104895B (en)
HR (1) HRP20100054T1 (en)
IL (1) IL179614A0 (en)
MA (1) MA28674B1 (en)
ME (1) ME01067B (en)
MX (1) MXPA06014228A (en)
NO (1) NO338097B1 (en)
NZ (1) NZ552220A (en)
PL (1) PL1831154T3 (en)
PT (1) PT1831154E (en)
RS (1) RS51210B (en)
SI (1) SI1831154T1 (en)
TW (6) TWI357405B (en)
UA (1) UA83575C2 (en)
WO (1) WO2006000904A2 (en)

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060276543A1 (en) * 2005-04-06 2006-12-07 Judith Aronhime Crystalline forms of pregabalin
US20060276544A1 (en) * 2005-05-10 2006-12-07 Lilach Hedvati Pregabalin free of isobutylglutaric acid and a process for preparation thereof
US20060281816A1 (en) * 2005-04-11 2006-12-14 Lilach Hedvati Pregabalin free of lactam and a process for preparation thereof
US20070043241A1 (en) * 2005-05-10 2007-02-22 Lilach Hedvati Optical resolution of 3-carbamoylmethyl-5-methylhexanoic acid
US20070066846A1 (en) * 2005-04-11 2007-03-22 Asher Maymon Process for making (S)-Pregabalin
US20070073085A1 (en) * 2005-05-10 2007-03-29 Lilach Hedvati Method for the preparation of pregabalin and salts thereof
CZ297970B6 (en) * 2005-08-10 2007-05-09 Zentiva, A. S Process for preparing (S)-3-(aminomethyl)-5-methyl-hexanoic acid (pregabalin)
US20070191636A1 (en) * 2005-09-19 2007-08-16 Kansal Vinod K Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-Pregabalin
US20070293694A1 (en) * 2006-05-24 2007-12-20 Lilach Hedvati Processes for the preparation of R-(+)-3-(carbamoyl methyl)-5-methylhexanoic acid and salts thereof
US20080015385A1 (en) * 2006-05-31 2008-01-17 Lilach Hedvati Preparation of (S)-pregabalin-nitrile
US20080026433A1 (en) * 2006-05-31 2008-01-31 Lilach Hedvati Use of enzymatic resolution for the preparation of intermediates of pregabalin
US20080306292A1 (en) * 2007-03-22 2008-12-11 Vinod Kumar Kansal Synthesis of (S)-(+)-3-(aminomethyl)-5-methyl hexanoic acid
US20080311635A1 (en) * 2007-05-14 2008-12-18 Dipharma Francis S.R.L. Process for the preparation of (s)(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090062393A1 (en) * 2007-08-29 2009-03-05 Protia, Llc Deuterium-enriched pregabalin
US20090137842A1 (en) * 2007-10-03 2009-05-28 Vollerner Yuri Pregabalin -4-eliminate, pregabalin 5-eliminate, their use as reference marker and standard, and method to produce pregabalin containing low levels thereof
US20090143615A1 (en) * 2007-12-03 2009-06-04 Dipharma Francis S.R.L. Process for the Preparation of (S)(+)-3-(Aminomethyl)-5-Methylhexanoic Acid
WO2009149928A1 (en) * 2008-06-10 2009-12-17 Synthon B.V. Processes for making pregabalin and intermediates therefor
US20100087525A1 (en) * 2008-06-23 2010-04-08 Lilach Hedvati Stereoselective enzymatic synthesis of (s) or (r)-iso-butyl-glutaric ester
WO2010070593A3 (en) * 2008-12-19 2010-11-04 Pfizer Ireland Pharmaceuticals Malonate esters
USRE41920E1 (en) 1996-07-24 2010-11-09 Warner-Lambert Company Llc Isobutylgaba and its derivatives for the treatment of pain
US20100312010A1 (en) * 2007-12-18 2010-12-09 Watson Pharma Private Limited Process for the Preparation of (S)-Pregabalin
WO2011141923A2 (en) 2010-05-14 2011-11-17 Lupin Limited Improved synthesis of optically pure (s) - 3-cyano-5-methyl-hexanoic acid alkyl ester, an intermediate of (s)- pregabalin
US8063244B2 (en) 2009-05-07 2011-11-22 Dipharma Francis S.R.L. Process for the synthesis of pregabalin
CN107445864A (en) * 2017-08-09 2017-12-08 无锡殷达尼龙有限公司 A kind of method for purifying Long carbon chain cyano group acid product
CN113651717A (en) * 2021-08-05 2021-11-16 浙江工业大学 A kind of racemization method of optically pure isobutyl succinonitrile

Families Citing this family (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5157576B2 (en) 2007-05-14 2013-03-06 住友化学株式会社 Process for producing optically active 2-alkyl-1,1,3-trialkoxycarbonylpropane
KR100846419B1 (en) * 2007-08-10 2008-07-15 한국과학기술원 New manufacturing method of pregabalin
JP2011507941A (en) * 2007-12-26 2011-03-10 ジェネリクス・（ユーケー）・リミテッド Method for producing pregabalin
SG157299A1 (en) * 2008-05-09 2009-12-29 Agency Science Tech & Res Diagnosis and treatment of kawasaki disease
CN102099482B (en) 2008-05-21 2014-04-16 桑多斯股份公司 Stereoselective enzymatic hydrolysis of 5-methyl-3-nitromethyl-hexanoate
WO2012025861A1 (en) 2010-08-23 2012-03-01 Pfizer Manufacturing Ireland Process for the preparation of ( s ) - 3 - cyano - 5 - methylhexanoic acid derivatives adn of pregabalin
WO2012059797A1 (en) * 2010-11-04 2012-05-10 Lupin Limited Process for synthesis of (s) - pregabalin
KR101306585B1 (en) * 2011-04-14 2013-09-10 한국외국어대학교 연구산학협력단 How to prepare pregabalin
CN102952057B (en) * 2012-11-07 2015-09-09 浙江普洛家园药业有限公司 A kind of preparation method of 4-isobutyl-pyrrolidin-2-one
JP6225490B2 (en) * 2013-05-31 2017-11-08 大日本印刷株式会社 Pharmaceutical package
US10076550B2 (en) * 2014-06-25 2018-09-18 Council Of Scientific & Industrial Research Synergistic pharmaceutical composition for gastroinestinal disorders
WO2016075082A1 (en) 2014-11-10 2016-05-19 Sandoz Ag Stereoselective reductive amination of alpha-chiral aldehydes using omega-transaminases for the synthesis of precursors of pregabalin and brivaracetam
CN104557576B (en) * 2014-12-19 2019-07-19 浙江华海药业股份有限公司 A kind of preparation method of high-purity Pregabalin
CN105348125A (en) * 2015-11-26 2016-02-24 太仓运通生物化工有限公司 Method for synthesizing Pregabalin by taking isovaleraldehyde as raw material
CN108358799B (en) * 2018-04-24 2020-11-10 贵州师范大学 Preparation method of pregabalin
CN109503403B (en) * 2018-12-21 2021-11-16 卓和药业集团股份有限公司 Pregabalin splitting method
CN113248416B (en) * 2020-02-10 2024-03-26 江西博腾药业有限公司 Preparation method and application of 1- ((benzyloxy) carbonyl) -4-ethylpyrrolidine-3-carboxylic acid
EP4103168A4 (en) * 2020-02-14 2024-04-24 Council Of Scientific & Industrial Research PROCESS FOR THE PRODUCTION OF GAMMA-AMINOBUTRY ACIDS AND THEIR ANALOGUES
CN111205288B (en) 2020-02-19 2021-03-30 四川大学 A kind of synthetic method of (1S,12bS) lactam ester compound
CN113880724A (en) * 2021-12-06 2022-01-04 南京桦冠生物技术有限公司 Preparation method of 3- (2-aminophenyl) -2-acrylate

Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5563175A (en) * 1990-11-27 1996-10-08 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5616793A (en) * 1995-06-02 1997-04-01 Warner-Lambert Company Methods of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US5618710A (en) * 1990-08-03 1997-04-08 Vertex Pharmaceuticals, Inc. Crosslinked enzyme crystals
US5637767A (en) * 1995-06-07 1997-06-10 Warner-Lambert Company Method of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US6001876A (en) * 1996-07-24 1999-12-14 Warner-Lambert Company Isobutylgaba and its derivatives for the treatment of pain
US6127418A (en) * 1997-08-20 2000-10-03 Warner-Lambert Company GABA analogs to prevent and treat gastrointestinal damage
US6194459B1 (en) * 1997-08-19 2001-02-27 Warner-Lambert Company Methods for treating physiological conditions associated with the use, or sequelae of use, of cocaine or other psychomotors stimulants
US6242488B1 (en) * 1997-08-20 2001-06-05 University Of Oklahoma Method for preventing and treating pain
US6306910B1 (en) * 1998-07-09 2001-10-23 Warner-Lambert Company Use of Gaba-analogues for treating insomnia
US6326374B1 (en) * 1998-07-09 2001-12-04 Warner-Lambert Company Compositions comprising GABA analogs and caffeine
US6329429B1 (en) * 1997-06-25 2001-12-11 Warner-Lambert Company Use of GABA analogs such as Gabapentin in the manufacture of a medicament for treating inflammatory diseases
US6359005B1 (en) * 1998-10-16 2002-03-19 Warner-Lambert Company Method for the treatment of mania and bipolar disorder
US20030149172A1 (en) * 2002-02-05 2003-08-07 Avantium International B.V. Crosslinked enzyme aggregates and crosslinking agent therefor
US20030212290A1 (en) * 2000-01-27 2003-11-13 Burk Mark Joseph Asymmetric synthesis of pregabalin

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3125583A (en) * 1964-03-17 Hjnc oxchaxohxchc o oh
US637767A (en) * 1899-09-25 1899-11-28 Gaylard W Denyes Grain-drill.
SU452196A1 (en) * 1973-02-12 2001-09-20 Филиал Всесоюзного Научно-Исследовательского Химико-Фармацевтического Института Им. Серго Орджоникидзе Method of producing γ-amino acid acid
JPS60338B2 (en) * 1980-12-16 1985-01-07 日本新薬株式会社 Production method of butyric acid derivatives
US4428887A (en) * 1982-07-14 1984-01-31 Monsanto Company Method of producing mono-substituted terminal diesters
DE19623142A1 (en) * 1996-06-10 1997-12-11 Huels Chemische Werke Ag Enantiomerically enriched malonic acid monoesters substituted by a tertiary hydrocarbon radical and their preparation
JPH10245369A (en) * 1997-03-03 1998-09-14 Ajinomoto Co Inc Production of serine derivative
RU2143002C1 (en) * 1997-12-24 1999-12-20 Акционерное общество открытого типа "Мосагроген" METHOD OF γ-AMINOBUTYRIC ACID PRODUCING
GB0004297D0 (en) * 2000-02-23 2000-04-12 Ucb Sa 2-oxo-1 pyrrolidine derivatives process for preparing them and their uses
US6931007B2 (en) * 2001-07-25 2005-08-16 Longboard, Inc. System and method of serving data messages
US20030225149A1 (en) * 2002-04-30 2003-12-04 Blazecka Peter G. Process for preparing highly functionalized gamma-butyrolactams and gamma-amino acids
FR2849024B1 (en) * 2002-12-20 2007-11-02 Aventis Pharma Sa NOVEL CHIRAL COMPOUNDS DERIVED FROM HEXANOIC ACID ESTERS, PROCESS AND PREPARATION INTERMEDIATES, USE IN THE SYNTHESIS OF CHIRAL 2- (BROMOMETHYL) 2-ETHYL HEXANOIC ACID
JP2006061112A (en) * 2004-08-30 2006-03-09 Sumitomo Chemical Co Ltd Process for producing optically active 2- (cyclopentylmethyl) -malonic acid monoester
KR20080036060A (en) * 2006-05-31 2008-04-24 테바 파마슈티컬 인더스트리즈 리미티드 How to Use Enzyme Splitting to Prepare Intermediates of Pregabalin

Patent Citations (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5618710A (en) * 1990-08-03 1997-04-08 Vertex Pharmaceuticals, Inc. Crosslinked enzyme crystals
US6028214A (en) * 1990-11-27 2000-02-22 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5599973A (en) * 1990-11-27 1997-02-04 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5608090A (en) * 1990-11-27 1997-03-04 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5563175A (en) * 1990-11-27 1996-10-08 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5684189A (en) * 1990-11-27 1997-11-04 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5710304A (en) * 1990-11-27 1998-01-20 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US6359169B1 (en) * 1990-11-27 2002-03-19 Northwestern University GABA and L-glutamic acid analogs for antiseizure treatment
US5847151A (en) * 1990-11-27 1998-12-08 Northwestern University Gaba and L-glutamic acid analogs for antiseizure treatment
US5616793A (en) * 1995-06-02 1997-04-01 Warner-Lambert Company Methods of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US5629447A (en) * 1995-06-02 1997-05-13 Warner-Lambert Company Methods of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US5637767A (en) * 1995-06-07 1997-06-10 Warner-Lambert Company Method of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US6046353A (en) * 1995-06-07 2000-04-04 Warner-Lambert Company Method of making (S)-3-(aminomethyl)-5-methylhexanoic acid
US5840956A (en) * 1995-06-07 1998-11-24 Warner-Lambert Company Method of making (S)-3-(Aminomethyl)-5-Methylhexanoic acid
US6001876A (en) * 1996-07-24 1999-12-14 Warner-Lambert Company Isobutylgaba and its derivatives for the treatment of pain
US6329429B1 (en) * 1997-06-25 2001-12-11 Warner-Lambert Company Use of GABA analogs such as Gabapentin in the manufacture of a medicament for treating inflammatory diseases
US6194459B1 (en) * 1997-08-19 2001-02-27 Warner-Lambert Company Methods for treating physiological conditions associated with the use, or sequelae of use, of cocaine or other psychomotors stimulants
US6242488B1 (en) * 1997-08-20 2001-06-05 University Of Oklahoma Method for preventing and treating pain
US6127418A (en) * 1997-08-20 2000-10-03 Warner-Lambert Company GABA analogs to prevent and treat gastrointestinal damage
US6426368B2 (en) * 1997-08-20 2002-07-30 Warner-Lambert Company Method for preventing and treating alcoholism
US6306910B1 (en) * 1998-07-09 2001-10-23 Warner-Lambert Company Use of Gaba-analogues for treating insomnia
US6326374B1 (en) * 1998-07-09 2001-12-04 Warner-Lambert Company Compositions comprising GABA analogs and caffeine
US6359005B1 (en) * 1998-10-16 2002-03-19 Warner-Lambert Company Method for the treatment of mania and bipolar disorder
US20030212290A1 (en) * 2000-01-27 2003-11-13 Burk Mark Joseph Asymmetric synthesis of pregabalin
US20030149172A1 (en) * 2002-02-05 2003-08-07 Avantium International B.V. Crosslinked enzyme aggregates and crosslinking agent therefor

Cited By (62)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE41920E1 (en) 1996-07-24 2010-11-09 Warner-Lambert Company Llc Isobutylgaba and its derivatives for the treatment of pain
US7417165B2 (en) 2005-04-06 2008-08-26 Teva Pharmaceutical Industries Ltd. Crystalline forms of pregabalin
US20060276543A1 (en) * 2005-04-06 2006-12-07 Judith Aronhime Crystalline forms of pregabalin
US20070066846A1 (en) * 2005-04-11 2007-03-22 Asher Maymon Process for making (S)-Pregabalin
US20060281816A1 (en) * 2005-04-11 2006-12-14 Lilach Hedvati Pregabalin free of lactam and a process for preparation thereof
US7488846B2 (en) 2005-04-11 2009-02-10 Teva Pharmaceuical Industries Ltd. Pregabalin free of lactam and a process for preparation thereof
US20070287860A1 (en) * 2005-05-10 2007-12-13 Lilach Hedvati Optical resolution of 3-carbamoylmethyl-5-methyl hexanoic acid
US7763749B2 (en) 2005-05-10 2010-07-27 Teva Pharmaceutical Industries Ltd. Method for the preparation of Pregabalin and salts thereof
US20070287748A1 (en) * 2005-05-10 2007-12-13 Lilach Hedvati Optical resolution of-3-carbamoylmethyl-5-methyl hexanoic acid
US20070043241A1 (en) * 2005-05-10 2007-02-22 Lilach Hedvati Optical resolution of 3-carbamoylmethyl-5-methylhexanoic acid
US20070287859A1 (en) * 2005-05-10 2007-12-13 Lilach Hedvati Optical resolution of 3-carbamoylmethyl-5-methyl hexanoic acid
US20060276544A1 (en) * 2005-05-10 2006-12-07 Lilach Hedvati Pregabalin free of isobutylglutaric acid and a process for preparation thereof
US7678938B2 (en) 2005-05-10 2010-03-16 Teva Pharmaceutical Industries Ltd. Optical resolution of 3-carbamoylmethyl-5-methyl hexanoic acid
US7619112B2 (en) 2005-05-10 2009-11-17 Teva Pharmaceutical Industries Ltd. Optical resolution of 3-carbamoylmethyl-5-methyl hexanoic acid
US20070073085A1 (en) * 2005-05-10 2007-03-29 Lilach Hedvati Method for the preparation of pregabalin and salts thereof
US7462737B2 (en) 2005-05-10 2008-12-09 Teva Pharmaceutical Industries Ltd. Pregabalin free of isobutylglutaric acid and a process for preparation thereof
CZ297970B6 (en) * 2005-08-10 2007-05-09 Zentiva, A. S Process for preparing (S)-3-(aminomethyl)-5-methyl-hexanoic acid (pregabalin)
US20100081841A1 (en) * 2005-09-19 2010-04-01 Vinod Kumar Kansal Novel asymmetric synthesis of (s)-(+)-3-(AMINOMETHYL)-5- methylhexanoic acid
US7923575B2 (en) 2005-09-19 2011-04-12 Teva Pharmaceutical Industries Limited Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7446220B2 (en) 2005-09-19 2008-11-04 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US8212071B2 (en) 2005-09-19 2012-07-03 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20080045747A1 (en) * 2005-09-19 2008-02-21 Kansal Vinod K Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-pregabalin
US7973196B2 (en) 2005-09-19 2011-07-05 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7465826B2 (en) 2005-09-19 2008-12-16 Teva Pharmaceutical Industries Ltd. Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-pregabalin
US7960583B2 (en) 2005-09-19 2011-06-14 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7470812B2 (en) 2005-09-19 2008-12-30 Teva Pharmaceutical Industries Ltd. Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-Pregabalin
US20080051602A1 (en) * 2005-09-19 2008-02-28 Kansal Vinod K Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-Pregabalin
US20090062562A1 (en) * 2005-09-19 2009-03-05 Vinod Kumar Kansal Novel asymmetric synthesis of (S)-(+)-3-(Aminomethyl)-5-methylhexanoic acid
US7851651B2 (en) 2005-09-19 2010-12-14 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090069578A1 (en) * 2005-09-19 2009-03-12 Vinod Kumar Kansal Novel asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090069600A1 (en) * 2005-09-19 2009-03-12 Vinod Kumar Kansal Novel asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090069596A1 (en) * 2005-09-19 2009-03-12 Vinod Kumar Kansal Novel asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090069593A1 (en) * 2005-09-19 2009-03-12 Vinod Kumar Kansal Novel asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US20070191636A1 (en) * 2005-09-19 2007-08-16 Kansal Vinod K Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-Pregabalin
US20070197827A1 (en) * 2005-09-19 2007-08-23 Kansal Vinod K Novel asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7687656B2 (en) 2005-09-19 2010-03-30 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7586005B2 (en) 2005-09-19 2009-09-08 Teva Pharmaceutical Industries Ltd. Asymmetric synthesis of (S)-(+)-3-(aminomethyl)-5-methylhexanoic acid
US7563923B2 (en) 2005-09-19 2009-07-21 Teva Pharmaceutical Industries Ltd. Chiral 3-carbamoylmethyl-5-methyl hexanoic acids, key intermediates for the synthesis of (S)-Pregabalin
US7462738B2 (en) 2006-05-24 2008-12-09 Teva Pharmaceutical Industries Ltd. Processes for the preparation of R-(+)-3-(carbamoyl methyl)-5-methylhexanoic acid and salts thereof
US20070293694A1 (en) * 2006-05-24 2007-12-20 Lilach Hedvati Processes for the preparation of R-(+)-3-(carbamoyl methyl)-5-methylhexanoic acid and salts thereof
EP2071032A2 (en) 2006-05-31 2009-06-17 Teva Pharmaceutical Industries Ltd The use of enzymatic resolution for the preparation of intermediates of pregabalin
US20080026433A1 (en) * 2006-05-31 2008-01-31 Lilach Hedvati Use of enzymatic resolution for the preparation of intermediates of pregabalin
US20080015385A1 (en) * 2006-05-31 2008-01-17 Lilach Hedvati Preparation of (S)-pregabalin-nitrile
WO2007143113A3 (en) * 2006-05-31 2008-01-31 Teva Pharma The use of enzymatic resolution for the preparation of intermediates of pregabalin
US8097754B2 (en) 2007-03-22 2012-01-17 Teva Pharmaceutical Industries Ltd. Synthesis of (S)-(+)-3-(aminomethyl)-5-methyl hexanoic acid
US20080306292A1 (en) * 2007-03-22 2008-12-11 Vinod Kumar Kansal Synthesis of (S)-(+)-3-(aminomethyl)-5-methyl hexanoic acid
US20080311635A1 (en) * 2007-05-14 2008-12-18 Dipharma Francis S.R.L. Process for the preparation of (s)(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090062393A1 (en) * 2007-08-29 2009-03-05 Protia, Llc Deuterium-enriched pregabalin
US20090137842A1 (en) * 2007-10-03 2009-05-28 Vollerner Yuri Pregabalin -4-eliminate, pregabalin 5-eliminate, their use as reference marker and standard, and method to produce pregabalin containing low levels thereof
EP2067768A1 (en) 2007-12-03 2009-06-10 Dipharma Francis S.r.l. A process for the preparation of (S)(+)-3-(aminomethyl)-5-methylhexanoic acid
US20090143615A1 (en) * 2007-12-03 2009-06-04 Dipharma Francis S.R.L. Process for the Preparation of (S)(+)-3-(Aminomethyl)-5-Methylhexanoic Acid
US20100312010A1 (en) * 2007-12-18 2010-12-09 Watson Pharma Private Limited Process for the Preparation of (S)-Pregabalin
US20110245508A1 (en) * 2008-06-10 2011-10-06 Lambertus Thijs Processes For Making Pregabalin And Intermediates Therefor
US20090312560A1 (en) * 2008-06-10 2009-12-17 Lambertus Thijs Processes for making pregabalin and intermediates therefor
WO2009149928A1 (en) * 2008-06-10 2009-12-17 Synthon B.V. Processes for making pregabalin and intermediates therefor
US20100087525A1 (en) * 2008-06-23 2010-04-08 Lilach Hedvati Stereoselective enzymatic synthesis of (s) or (r)-iso-butyl-glutaric ester
WO2010070593A3 (en) * 2008-12-19 2010-11-04 Pfizer Ireland Pharmaceuticals Malonate esters
US8063244B2 (en) 2009-05-07 2011-11-22 Dipharma Francis S.R.L. Process for the synthesis of pregabalin
WO2011141923A2 (en) 2010-05-14 2011-11-17 Lupin Limited Improved synthesis of optically pure (s) - 3-cyano-5-methyl-hexanoic acid alkyl ester, an intermediate of (s)- pregabalin
WO2011141923A3 (en) * 2010-05-14 2012-01-19 Lupin Limited Improved synthesis of optically pure (s) - 3-cyano-5-methyl-hexanoic acid alkyl ester, an intermediate of (s)- pregabalin
CN107445864A (en) * 2017-08-09 2017-12-08 无锡殷达尼龙有限公司 A kind of method for purifying Long carbon chain cyano group acid product
CN113651717A (en) * 2021-08-05 2021-11-16 浙江工业大学 A kind of racemization method of optically pure isobutyl succinonitrile

Also Published As

Publication number Publication date
KR100847928B1 (en) 2008-07-22
NO338097B1 (en) 2016-08-01
ES2336014T3 (en) 2010-04-07
US8044227B2 (en) 2011-10-25
TWI402248B (en) 2013-07-21
EA200602099A1 (en) 2007-04-27
GEP20104895B (en) 2010-02-25
CN102102114A (en) 2011-06-22
CR8765A (en) 2008-01-02
CN102102114B (en) 2013-08-14
TW200604139A (en) 2006-02-01
US20120015412A1 (en) 2012-01-19
ATE455093T1 (en) 2010-01-15
CA2571040A1 (en) 2006-01-05
BRPI0512347A (en) 2008-03-04
JP4782101B2 (en) 2011-09-28
HK1161307A1 (en) 2012-08-24
TW200900375A (en) 2009-01-01
EP1831154A2 (en) 2007-09-12
TWI377191B (en) 2012-11-21
SI1831154T1 (en) 2010-03-31
EA200800908A1 (en) 2008-12-30
PL1831154T3 (en) 2010-05-31
US8134023B2 (en) 2012-03-13
KR20080015152A (en) 2008-02-18
BRPI0512347B1 (en) 2018-03-20
AU2005256945B2 (en) 2008-11-13
JP2008088191A (en) 2008-04-17
CN1972904B (en) 2011-02-02
JP4966183B2 (en) 2012-07-04
KR100847927B1 (en) 2008-07-22
MA28674B1 (en) 2007-06-01
WO2006000904A3 (en) 2006-04-27
NO20065329L (en) 2006-12-19
AP2006003846A0 (en) 2006-12-31
BRPI0512347B8 (en) 2021-05-25
TW200846306A (en) 2008-12-01
EA015418B1 (en) 2011-08-30
TWI405747B (en) 2013-08-21
US20110065168A1 (en) 2011-03-17
ME01067B (en) 2012-10-20
KR100847929B1 (en) 2008-07-22
TWI399361B (en) 2013-06-21
CA2571040C (en) 2009-09-15
JP2009022272A (en) 2009-02-05
AU2005256945A1 (en) 2006-01-05
JP2008133289A (en) 2008-06-12
HRP20100054T1 (en) 2010-05-31
EA011765B1 (en) 2009-06-30
DE602005018965D1 (en) 2010-03-04
JP2008110985A (en) 2008-05-15
EA200800909A1 (en) 2008-08-29
TW200846307A (en) 2008-12-01
MXPA06014228A (en) 2007-02-14
JP4777332B2 (en) 2011-09-21
TWI357405B (en) 2012-02-01
CR20110483A (en) 2011-10-24
TW200846474A (en) 2008-12-01
KR20080015524A (en) 2008-02-19
JP4800346B2 (en) 2011-10-26
HK1104024A1 (en) 2008-01-04
CY1110337T1 (en) 2015-01-14
RS51210B (en) 2010-12-31
CN1972904A (en) 2007-05-30
NZ552220A (en) 2010-08-27
US7838686B2 (en) 2010-11-23
TWI396743B (en) 2013-05-21
TW200902481A (en) 2009-01-16
DK1831154T3 (en) 2010-04-12
US20090042262A1 (en) 2009-02-12
AP2466A (en) 2012-09-17
EP1831154B1 (en) 2010-01-13
WO2006000904A2 (en) 2006-01-05
UA83575C2 (en) 2008-07-25
AR049446A1 (en) 2006-08-02
IL179614A0 (en) 2007-05-15
JP2008504252A (en) 2008-02-14
JP4174551B2 (en) 2008-11-05
PT1831154E (en) 2010-02-19
KR20080015153A (en) 2008-02-18

Similar Documents

Publication Publication Date Title
US8044227B2 (en) Preparation of pregabalin and related compounds
US20070141684A1 (en) Preparation of gamma-amino acids having affinity for the alpha-2-delta protein
KR100843684B1 (en) Process for preparing pregabalin and related compounds
HK1104024B (en) Preparation of pregabalin and related compounds
HK1161307B (en) Preparation of pregabalin and related compounds

Legal Events

Date Code Title Description
STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION