WO2009005647A2 - Composés et procédé de préparation d'intermédiaires chiraux pour la synthèse de la proxétine - Google Patents

Composés et procédé de préparation d'intermédiaires chiraux pour la synthèse de la proxétine Download PDF

Info

Publication number
WO2009005647A2
WO2009005647A2 PCT/US2008/007870 US2008007870W WO2009005647A2 WO 2009005647 A2 WO2009005647 A2 WO 2009005647A2 US 2008007870 W US2008007870 W US 2008007870W WO 2009005647 A2 WO2009005647 A2 WO 2009005647A2
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
haloalkyl
heteroalkyl
alkynyl
alkenyl
Prior art date
Application number
PCT/US2008/007870
Other languages
English (en)
Other versions
WO2009005647A3 (fr
Inventor
Robert William Scott
Sean Timothy Neville
Lishan Zhao
Ningqing Ran
Original Assignee
Bioverdant, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Bioverdant, Inc. filed Critical Bioverdant, Inc.
Publication of WO2009005647A2 publication Critical patent/WO2009005647A2/fr
Publication of WO2009005647A3 publication Critical patent/WO2009005647A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms

Definitions

  • Paroxetine is a drug that treats a variety of conditions such as anti-depression, anxiety, panic disorders, post-traumatic stress disorder, premenstrual dysphoric disorder, and obsessive-compulsive disorders (Lund et al., Acta Pharmacol. Toxicol. 51, 351, 1982; Zohar et al, Brit. J. Psychiatry 169, 468, 1996; Stein et al, J. Am. Med. Assoc. 280, 708, 1998; Dechant et al, Drugs, 41, 225, 1991 ; and Dunner et al., Pharmacopsychiatry 31 , 89, 1998).
  • Paroxetine also is used for treating chronic headaches, tingling in the hands and feet, certain male sexual problems and bipolar disorders. Accordingly, there is high demand for paroxetine compounds.
  • the method includes enzymatic hydrolysis of a piperidine derivative, such as meso-(3R,4s,5S)-dimethyl 4-(4-fluorophenyl)-l- methyl-2,6-dioxopiperidine-3,5-dicarboxylate, resulting in the formation of a trans- 3,4-disubstituted piperidine derivative, which is converted to paroxetine via subsequent mesylation, displacement with sesamol, and demethylation.
  • a piperidine derivative such as meso-(3R,4s,5S)-dimethyl 4-(4-fluorophenyl)-l- methyl-2,6-dioxopiperidine-3,5-dicarboxylate
  • R 1 is selected from among hydrogen, OR', NR 'R", Ci- C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, C]-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR',
  • R 1 is selected from among hydrogen, Ci-C 8 alkyl, C 2 - C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl; and X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, C 1 -C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 -
  • R 1 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 haloalkyl, Cj-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 7 cycloalkyl, C 4 -C 7 heterocycloalkyl having one to five heteroatoms selected from among O, N, S and P, phenyl and benzyl, wherein the alkyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, phenyl and benzyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C 6 - haloalkyl, Ci-C 6 -C 6 -
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , CpC 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl.
  • X is halogen.
  • X is F, Cl, Br or I.
  • X is CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R' or NR'R", wherein R' and R" each independently is selected from among hydrogen, Cj- C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • X is F. In certain embodiments X is 4-fluoro.
  • R 1 is selected from among hydrogen, OR', NR'R", C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, C 1 -C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", CpC 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C]-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted; and R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Cj-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl; and
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Cj-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Cj-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 - C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N, S and P.
  • R 1 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 7 cycloalkyl, C 4 -C 7 heterocycloalkyl having one to five heteroatoms selected from among O, N, S and P, phenyl and benzyl, wherein the alkyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, phenyl and benzyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C 6 - haloalkyl, Ci-C 6 -hydroxy
  • R 2 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Cj-C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 7 cycloalkyl, C 4 -C 6 heterocycloalkyl having one to five heteroatoms selected from among O, N, S and P, phenyl and benzyl, wherein the alkyl, alkenyl, alkynyl haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, phenyl and benzyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, C]-C 6 -alkyl, Ci-C 6 -haloalkyl,
  • X is halogen. In certain embodiments X is F, Cl, Br or I.
  • X is CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R' or NR'R", wherein R' and R" each independently is selected from among hydrogen, Cj- C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, CpC 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • X is F. In some embodiments X is 4-fluoro.
  • the process includes the step of: reacting a compound of Formula II:
  • a R 1 is selected from among OR', NR'R", C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C r C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, CpC 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 hetero alkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • A is selected from among F, Cl, Br, I, CN, NO 2 , OR', SR', SOR' and SO 2 R'.
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -
  • substituents can be selected from among a subset of the listed alternatives.
  • the process includes reacting a compound of Formula IV: with a compound of Formula V:
  • R 1 is selected from among hydrogen, OR', NR 'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • R' and R" each independently is selected from among hydrogen, Cj-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • the process includes the step of: transesterifying a compound of Formula I:
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • R 2 and R 3 each independently is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, C)-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives. Also provided is a process for producing a compound of Formula VII:
  • the process includes the step of: hydrolyzing a compound of Formula I:
  • I R 1 is selected from among hydrogen, OR', NR'R", C r C 8 alkyl, C 2 -C 8 alkenyl,
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO2R', CO2R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C r C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 - C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • R' and R" each independently is selected from among hydrogen, C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • processes for producing compounds of Formula VII comprising esterase-catalyzed hydrolysis of compounds of Formula I.
  • processes for producing compounds of Formula VII comprising protease-catalyzed hydrolysis of compounds of Formula I.
  • the processes include enzymatic hydrolysis of compounds of Formula I.
  • the enzyme used in the process is a hydrolase.
  • the enzyme is an esterase or a protease.
  • the enzyme is selected from among Lipase ASl, Pig liver esterase, Rabbit liver esterase, Protease P, Protease M, Subtilisin A, ALCALASE ® , OPTIMASE ® , Alkaline protease, VALIDASE ® , Protease Type XXIV, Proteinase and Subtilisin Carlsberg.
  • the enzyme can be selected from among a carboxylesterase of SEQ ID NOS: 1-3 and 28-113.
  • the enzyme can be selected from among a subtilisin of SEQ ID NOS:4-8 and 15-27.
  • the enzyme can be selected from among an oryzin of SEQ ID NOS:9-14.
  • the enzyme can be selected from among an endopeptidase of SEQ ID NOS:114-172.
  • the enzyme can be selected from among a lipase of SEQ ID NOS:173-187.
  • the compounds produced by the methods provided herein have an optical purity of between 65.0-99.9%. In other embodiments, the optical purity of the compounds of Formula VII is between 75-99%. In other embodiments, the optical purity of the compounds of Formula VII is between 90-95%.
  • the process includes the steps of: reacting a compound of Formula I:
  • R 1 is selected from among hydrogen, OR', NR'R", C r C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Cj-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Cj-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C r C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • R' and R" each independently is selected from among hydrogen, Cj-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • Provided herein is a process for producing paroxetine and related compounds.
  • the process includes the steps of:
  • step (d) reacting the activated compound of step (c) with a sesamol derivative to produce a compound of Formula IX:
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, CpC 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Cj-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci -C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Cj-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted.
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -Cg alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • a mesylating reagent is a compound of the formula R'-SO 2 X where R' is alkyl, haloalkyl, heteroalkyl, heteroaryl or aryl and X is F, Cl, Br or I.
  • the mesylating reagent is a compound of the formula SOX 2 where X is F, Cl, Br or I.
  • the mesylating reagent is selected from among sulfonyl chloride, p-toluenesulfonyl chloride, p-bromobenzensulfonyl chloride, p- nitrobenzyenesulfonyl chloride, methanesulfonyl chloride, trifluoromethanesulfonyl chloride, nonafluorobutanesulfonyl chloride, and 2,2,2-trifluoroethanesulfonyl chloride.
  • the mesylating reagent is methanesulfonyl chloride.
  • the activated compound from step (c) is reacted with sesamol. In other embodiments, the activated compound from step (c) is reacted with benzo[d][l,3]dioxol-5-olate.
  • an activated sesamol derivative is formed by reacting sesamol with a base such as potassium t-butoxide, lithium diisopropylamide, butyllithium, alkali metal hydrides such as potassium hydride, alkali metals such as sodium, or metal chlorides such as sodium chloride to form an activated sesamol derivative.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 - C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein R' and R" each independently is selected from among hydrogen, C)-C 8 alkyl, C 2 -Cg alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, CpC 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alken
  • substituents can be selected from among a subset of the listed alternatives.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , C r C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 - C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamid
  • articles of manufacture that include packaging material, an intermediate compound provided herein, such as a compound of Formula I or a pharmaceutically acceptable salt thereof within the packaging material, and a label that indicates that the compound is used for the synthesis of paroxetine.
  • kits for the synthesis of paroxetine that include a hydrolase and an intermediate compound as provided herein or a pharmaceutically acceptable salt thereof.
  • Standard techniques can be used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation and delivery, and treatment of subjects.
  • Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection).
  • Reactions and purification techniques can be performed e.g., using kits o/manufacturer's specifications or as commonly accomplished in the art or as described herein.
  • the foregoing techniques and procedures generally are performed of conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g. , Sambrook et ah, Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989).
  • treating encompass either or both responsive and prophylaxis measures, e.g., designed to inhibit, slow or delay the onset of a symptom of a disease or disorder, achieve a full or partial reduction of a symptom or disease state, and/or to alleviate, ameliorate, lessen, or cure a disease or disorder and/or its symptoms.
  • C 1 -C x includes Ci-C 2 , CpC 3 . . . Ci-C x.
  • alkyl refers to straight or branched chain substituted or unsubstituted hydrocarbon groups, in one embodiment 1 to 40 carbon atoms, in another embodiment, 1 to 20 carbon atoms, in another embodiment, 1 to 10 carbon atoms.
  • lower alkyl refers to alkyl groups of 1 to 6 carbon atoms.
  • An alkyl group can be a "saturated alkyl,” meaning that it does not contain any alkene or alkyne groups and in certain embodiments, alkyl groups are optionally substituted.
  • An alkyl group can be an "unsaturated alkyl,” meaning that it contains at least one alkene or alkyne group.
  • An alkyl group that includes at least one carbon- carbon triple bond (C ⁇ €) also is referred to by the term "alkynyl,” and in certain embodiments, alkynyl groups are optionally substituted.
  • an alkyl contains 1 to 20 carbon atoms (whenever it appears herein, a numerical range such as “1 to 20” refers to each integer in the given range; e.g., "1 to 20 carbon atoms” means that an alkyl group can contain only 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms, although the term “alkyl” also includes instances where no numerical range of carbon atoms is designated).
  • An alkyl can be designated as "Ci-C 4 alkyl" or by similar designations.
  • C 1 -C 4 alkyl indicates an alkyl having one, two, three, or four carbon atoms, i.e., the alkyl is selected from among methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl and t-butyl.
  • Ci - C 4 includes Ci - C 2 , Cj - C 3 , C 2 - C 3 and C 2 - C 4 alkyl.
  • Alkyls include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, ethenyl, propenyl, butenyl, hexenyl, ethynyl, propynyl, butynyl and hexynyl.
  • halogen or “halide” refers to F, Cl, Br or I and includes pseudohalides.
  • pseudohalides are compounds that behave substantially similar to halides. Such compounds can be used in the same manner and treated in the same manner as halides (X-, in which X is a halogen, such as Cl, F or Br).
  • Pseudohalides include, but are not limited to, cyanide, cyanate, thiocyanate, selenocyanate, trifluoromethoxy, trifluoromethyl and azide.
  • haloalkyl alone or in combination refers to an alkyl in which at least one hydrogen atom is replaced with a halogen atom.
  • the halogen atoms are all the same as one another. In certain of such embodiments, the halogen atoms are not all the same as one another.
  • Certain haloalkyls are saturated haloalkyls, which do not include any carbon-carbon double bonds or any carbon- carbon triple bonds.
  • Certain haloalkyls are haloalkenes, which include one or more carbon-carbon double bonds.
  • Certain haloalkyls are haloalkynes, which include one or more carbon-carbon triple bonds. In certain embodiments, haloalkyls are optionally substituted. Where the number of any given substiruent is not specified (e.g.
  • haloalkyl there can be one or more substituents present.
  • haloalkyl can include one or more of the same or different halogens.
  • haloalkyl includes each of the substituents CF 3 , CHF 2 and CH 2 F.
  • heteroatom refers to an atom other than carbon or hydrogen. Heteroatoms are typically independently selected from oxygen, sulfur, nitrogen, and phosphorus, but are not limited to those atoms. In embodiments in which two or more heteroatoms are present, the two or more heteroatoms can all be the same as one another, or some or all of the two or more heteroatoms can each be different from the others.
  • heteroalkyl alone or in combination refers to a group containing an alkyl and one or more heteroatoms. Certain heteroalkyls are saturated heteroalkyls, which do not contain any carbon-carbon double bonds or any carbon-carbon triple bonds.
  • heterohaloalkyl alone or in combination refers to a heteroalkyl in which at least one hydrogen atom is replaced with a halogen atom, hi certain embodiments, heteroalkyls are optionally substituted.
  • non-cyclic alkyl refers to an alkyl that is not cyclic (i.e., a straight or branched chain containing at least one carbon atom).
  • Non-cyclic alkyls can be fully saturated or can contain non-cyclic alkenes and/or alkynes.
  • Non- cyclic alkyls can be optionally substituted.
  • ring refers to any covalently closed structure.
  • Rings include, for example, carbocycles (e.g., aryls and cycloalkyls), heterocycles (e.g., heteroaryls and non-aromatic heterocycles), aromatics (e.g., aryls and heteroaryls), and non-aromatics (e.g., cycloalkyls and non-aromatic heterocycles). Rings can be optionally substituted. Rings can form part of a ring system. As used herein, the term “ring system” refers to two or more rings, wherein two or more of the rings are fused. The term “fused" refers to structures in which two or more rings share one or more bonds.
  • carbocycles e.g., aryls and cycloalkyls
  • heterocycles e.g., heteroaryls and non-aromatic heterocycles
  • aromatics e.g., aryls and heteroaryls
  • non-aromatics e.g
  • Carbocycle refers to a ring where each of the atoms forming the ring is a carbon atom.
  • Carbocyclic rings can be formed by three, four, five, six, seven, eight, nine, or more than nine carbon atoms.
  • Carbocycles can be optionally substituted.
  • cycloalkyl refers to a saturated mono- or multicyclic ring system where each of the atoms forming a ring is a carbon atom. Cycloalkyls can be formed by three, four, five, six, seven, eight, nine, or more than nine carbon atoms, hi one embodiment, the ring system includes 3 to 12 carbon atoms. In another embodiment, they ring system includes 3 to 6 carbon atoms.
  • the term "cycloalkyl” includes rings that contain one or more unsaturated bonds. As used herein, the terms “cycloalkenyl” and “cycloalkynyl” are unsaturated cycloalkyl ring system. Cycloalkyls can be optionally substituted.
  • a cycloalkyl contains one or more unsaturated bonds.
  • cycloalkyls include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclopentadiene, cyclohexane, cyclohexene, 1,3-cyclohexadiene, 1 ,4-cyclohexadiene, cycloheptane and cycloheptene.
  • cycloalkynyl refers to mono- or multicyclic ring systems that includes at least one carbon-carbon triple bond (C ⁇ C). Cycloalkenyl and cycloalkynyl groups include ring systems that include 3 to
  • the cycloalkenyl groups include 4 to 7 carbon atoms.
  • the cycloalkynyl groups include 8 to 10 carbon atoms.
  • the ring systems of the cycloalkyl, cycloalkenyl and cycloalkynyl groups can be composed of one ring or two or more rings that can be joined together in a fused, bridged or spiro-connected fashion, and can be optionally substituted with one or more alkyl group substituents.
  • heterocycle refers to a ring wherein at least one atom forming the ring is a carbon atom and at least one atom forming the ring is a heteroatom.
  • Heterocyclic rings can be formed by three, four, five, six, seven, eight, nine, or more than nine atoms. Any number of those atoms can be heteroatoms (i.e., a heterocyclic ring can contain one, two, three, four, five, six, seven, eight, nine, or more than nine heteroatoms, provided that at lease one atom in the ring is a carbon atom).
  • the heterocyclic ring will have additional heteroatoms in the ring.
  • 4-6 membered heterocycle refer to the total number of atoms that comprise the ring (i.e., a four, five, or six membered ring, in which at least one atom is a carbon atom, at least one atom is a heteroatom and the remaining two to four atoms are either carbon atoms or heteroatoms).
  • those two or more heteroatoms can be the same or different from one another.
  • the heterocycle includes 3-12 members.
  • the heterocycle includes 4, 5, 6, 7 or 8 members.
  • the heterocycle can be optionally substituted with one or more substituents.
  • the substituents of the heterocyclic group are selected from among hydroxy, amino, alkoxy containing 1 to 4 carbon atoms, halo lower alkyl, including trihalomethyl, such as trifluoromethyl, and halogen.
  • heterocycle can include reference to heteroaryl. Binding to a heterocycle can be at a heteroatom or via a carbon atom. Examples of heterocycles include, but are not limited to the following:
  • D, E, F and G independently represent a heteroatom.
  • D, E, F and G can be the same or different from one another.
  • bicyclic ring refers to two rings, wherein the two rings are fused.
  • Bicyclic rings include, for example, decaline, pentalene, naphthalene, azulene, heptalene, isobenzofuran, chromene, indolizine, isoindole, indole, purine, indoline, indene, quinolizine, isoquinoline, quinoline, phthalazine, naphthyrididine, quinoxaline, cinnoline, pteridine, isochroman, chroman and various hydrogenated derivatives thereof.
  • Bicyclic rings can be optionally substituted. Each ring is independently aromatic or non-aromatic.
  • both rings are aromatic. In certain embodiments, both rings are non-aromatic. In certain embodiments, one ring is aromatic and one ring is non-aromatic.
  • aromatic refers to a planar ring having a delocalized ⁇ -electron system containing 4n+2 ⁇ electrons, where n is an integer.
  • Aromatic rings can be formed by five, six, seven, eight, nine, or more than nine atoms. Aromatics can be optionally substituted.
  • aromatic groups include, but are not limited to phenyl, naphthalenyl, phenanthrenyl, anthracenyl, tetralinyl, fluorenyl, indenyl and indanyl.
  • aromatic includes, for example, benzenoid groups, connected via one of the ring- forming carbon atoms, and optionally carrying one or more substituents selected from an aryl, a heteroaryl, a cycloalkyl, a non-aromatic heterocycle, a halo, a hydroxy, an amino, a cyano, a nitro, an alkylamido, an acyl, a C] -6 alkoxy, a C 1-6 alkyl, a C J-6 hydroxyalkyl, a Ci -6 aminoalkyl, a C 1-6 alkylamino, an alkylsulfenyl, an alkylsulfinyl, an alkylsulfonyl,
  • an aromatic group is substituted at one or more of the para, meta, and/or ortho positions.
  • aromatic groups containing substitutions include, but are not limited to, phenyl, 3-halophenyl, 4-halophenyl, 3-hydroxyphenyl, 4-hydroxy- phenyl, 3-aminophenyl, 4-aminophenyl, 3-methylphenyl, 4-methylphenyl, 3- methoxyphenyl, 4-methoxyphenyl, 4-trifluoromethoxyphenyl, 3-cyano-phenyl, 4- cyanophenyl, dimethylphenyl, naphthyl, hydroxynaphthyl, hydroxymethyl-phenyl, (trifiuoromethyl)phenyl, alkoxyphenyl, 4-morpholin-4-ylphenyl, 4-pyrrolidin-l- ylphenyl, 4-pyrazolylphenyl, 4-triazolylphenyl and 4-(2-oxopyrrolidin-l-yl-yl
  • aryl refers to an aromatic ring wherein each of the atoms forming the ring is a carbon atom.
  • Aryl rings can be formed by three, four, five, six, seven, eight, nine, or more than nine carbon atoms.
  • Aryl groups can be optionally substituted.
  • heteroaryl refers to an aromatic ring in which at least one atom forming the aromatic ring is a heteroatom.
  • Heteroaryl rings can be formed by three, four, five, six, seven, eight, nine and more than nine atoms.
  • Heteroaryl groups can be optionally substituted.
  • heteroaryl groups include, but are not limited to, aromatic C 3-8 heterocyclic groups containing one oxygen or sulfur atom, or two oxygen atoms, or two sulfur atoms or up to four nitrogen atoms, or a combination of one oxygen or sulfur atom and up to two nitrogen atoms, and their substituted as well as benzo- and pyrido-fused derivatives, for example, connected via one of the ring-forming carbon atoms.
  • heteroaryl groups are optionally substituted.
  • the one or more substituents are each independently selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, C t - 6 -alkoxy, Ci -6 -alkyl, Ci -6 -haloalkyl, Ci -6 -hydroxy-alkyl, Ci -6 -aminoalkyl, C 1-6 -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl, or trifluoromethyl.
  • heteroaryl groups include, but are not limited to, unsubstituted and mono- or di- substituted derivatives of furan, benzo furan, thiophene, benzothiophene, pyrrole, pyridine, indole, oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole, isothiazole, imidazole, benzimidazole, pyrazole, indazole, tetrazole, quinoline, isoquinoline, pyridazine, pyrimidine, purine and pyrazine, furazan, 1,2,3- oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, triazole, benzotriazole, pteridine, phenoxazole, oxadiazole, benzopyrazole, quinolizine, cinnoline, phthalazine,
  • the substituents are halo, hydroxy, cyano, O-C 1-6 -alkyl, Ci -6 -alkyl, hydroxy-C 1-6 -alkyl and amino-Ci -6 -alkyl.
  • non-aromatic ring refers to a ring that does not have a delocalized 4n+2 ⁇ -electron system.
  • non-aromatic heterocycle refers to a non-aromatic ring wherein one or more atoms forming the ring is a heteroatom.
  • Non-aromatic heterocyclic rings can be formed by three, four, five, six, seven, eight, nine, or more than nine atoms.
  • Non-aromatic heterocycles can be optionally substituted.
  • non-aromatic heterocycles contain one or more carbonyl or thiocarbonyl groups such as, for example, oxo- and thio-containing groups.
  • non- aromatic heterocycles include, but are not limited to, lactams, lactones, cyclic imides, cyclic thioimides, cyclic carbamates, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin, 1,3-dioxane, 1,4-dioxin, 1 ,4-dioxane, piperazine, 1,3-oxathiane, 1 ,4-oxathiin, 1 ,4-oxathiane, tetrahydro-l,4-thiazine, 2//-l,2-oxazine , maleimide, succinimide, barbituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane, hexahydro-l,3,5-triazine, te
  • arylalkyl alone or in combination, refers to an alkyl substituted with an aryl that can be optionally substituted.
  • heteroarylalkyl alone or in combination, refers to an alkyl substituted with a heteroaryl that can be optionally substituted.
  • cyano refers to a group of formula -CN.
  • isocyanato refers to a group of formula -NCO.
  • thiocyanato refers to a group of formula -CNS.
  • isothiocyanato refers to a group of formula -NCS.
  • esters refers to a chemical moiety with formula -(R) n -COOR', where R and R' each independently is selected from among alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and non-aromatic heterocycle (bonded through a ring carbon), where n is 0 or 1.
  • amide refers to a chemical moiety with formula -(R) n -C(O)NHR' or -(R) n -NHC(O)R', where R and R' each independently is selected from alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic (bonded through a ring carbon), where n is 0 or 1.
  • R and R' each independently is selected from alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) and heteroalicyclic (bonded through a ring carbon), where n is 0 or 1.
  • an amide can be an amino acid or a peptide.
  • amine As used herein, the terms “amine,” “hydroxy,” and “carboxyl” include such groups that have been esterified or amidif ⁇ ed. Procedures and specific groups used to achieve esterification and amidification are known to those of skill in the art and can readily be found in reference sources such as Greene and Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., John Wiley & Sons, New York, NY, 1999, which is incorporated herein in its entirety, where permitted. As used herein, the term “together form a bond” refers to the instance in which two substituents to neighboring atoms are null the bond between the neighboring atoms becomes a double bond.
  • the term “optionally substituted,” refers to a group in which none, one, or more than one of the hydrogen atoms has been replaced with one or more group(s) individually and independently selected from among alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl, non-aromatic heterocycle, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N carbamyl, O thiocarbamyl, N thiocarbamyl, C amido, N amido, S-sulfonamido, N sulfonamido, C carboxy, O carboxy, isocyanato, thiocyanato, isothiocyanato, nitro, silyl, trihalomethan
  • the compounds provided herein can contain chiral centers. Such chiral centers can be of either the (R) or (S) configuration, or can be a mixture thereof. Thus, the compounds provided herein can be enantiomerically pure, or be stereoisomeric or diastereomeric mixtures. It is to be understood that the chiral centers of the compounds provided herein can undergo epimerization in vivo. As such, one of skill in the art will recognize that administration of a compound in its (R) form is equivalent, for compounds that undergo epimerization in vivo, to administration of the compound in its (S) form.
  • cis and trans are descriptors that show the relationship between two ligands attached to separate atoms that are connected by a double bond or contained in a ring with a double bond.
  • Two ligands are said to be “cis” to each other if they lie on the same side of a plane. If the ligands are on opposite sides, their relative position is described as “trans.”
  • the appropriate reference plane of a double bond is perpendicular to that of the relevant sigma bond that passes through the double bond.
  • enantiomer refers to one of a pair of molecular entities that are mirror images of each other and non-superimposable.
  • Enantiomeric excess (ee) can be calculated for a mixture of (R) and (S)-enantiomers.
  • the ee is defined as the absolute value of the mole fractions of F ⁇ minus the mole fraction of F (S y
  • the percent ee is the absolute value of the mole fractions of F ⁇ minus the mole fraction of F ( s ) multiplied by 100.
  • optical activity refers to the ability of a sample material to rotate the plane of polarized light. A specific enantiomer causes rotation of light in either a clockwise or counterclockwise direction.
  • optical purity refers to the ratio of observed optical rotation of a sample comprising a mixture of enantiomers to the optical rotation of one pure enantiomer.
  • substantially pure means sufficiently homogeneous to appear free of readily detectable impurities as determined by standard methods of analysis, such as thin layer chromatography (TLC), gel electrophoresis, high performance liquid chromatography (HPLC) and mass spectrometry (MS), used by those of skill in the art to assess such purity, or sufficiently pure such that further purification would not detectably alter the physical and chemical properties, such as enzymatic and biological activities, of the substance.
  • substantially pure object species e.g., compound
  • is the predominant species present i.e., on a molar basis it is more abundant than any other individual species in the composition.
  • a substantially purified fraction is a composition wherein the object species contains at least about 50 percent (on a molar basis) of all species present. In certain embodiments, a substantially pure composition will contain more than about 50%, 60%, 70%, 80%, 85%, 90%, 95%, or 99% of all species present in the composition. In certain embodiments, a substantially pure composition will contain more than about 80%, 85%, 90%, 95%, or 99% of all species present in the composition.
  • Methods for purification of the compounds to produce substantially chemically pure compounds are known to those of skill in the art. A substantially chemically pure compound can, however, be a mixture of stereoisomers. In such instances, further purification might increase the specific activity of the compound.
  • Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as reverse phase HPLC.
  • the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.
  • acid generally refers to a molecular entity or chemical species capable of donating a hydron (proton), otherwise considered a Br ⁇ nsted acid, or capable of accepting an electron pair, otherwise known as a Lewis acid.
  • base refers to a chemical species or molecular entity having an available pair of electrons capable of forming a bond with a hydron (proton), otherwise known as a Br ⁇ nsted base, or capable of donating an electron pair to form a bond with the vacant orbital of some other species, otherwise known as a Lewis base.
  • activation of a chemical group occurs when some or all of the energy required for a desired transformation is provided by a preceding reaction. For example, in the scheme: A + B ⁇ X ⁇ Y + Z, some or all of the energy required for the reaction of X to form products Y and Z is provided by the first reaction between A and B.
  • chemical resolution occurs when a mixture of stereoisomers is separated into the component diasteriomers and/or enantiomers. Chemical resolution also can be applied to the separation of olefin cis- and trans- isomers.
  • an "aliquot” refers to a fractional part of known quantity taken from a larger solution or mixture. The properties of the aliquot are usually analyzed and such properties are considered to be representative of the properties of the larger solution or mixture.
  • a "reactant” refers to a substance that is consumed in the course of a chemical reaction. A reactant also is referred to as a reagent.
  • a "condensation” reaction occurs when two or more reactants yield a single main product with accompanying formation of water or of some other small molecule such as ammonia, ethanol, acetic acid, or hydrogen sulfide. A condensation reaction also can occur between two or more reactive sites within the same molecular entity.
  • transesterification refers to a reaction that converts one ester into another. Transesterifications are often realized by reacting an ester with an excess of alcohol under acidic or basic conditions.
  • transformation refers to the conversion of a substrate into a particular product irrespective of the reagents or mechanisms involved. Reference to a transformation does not require full description of all reactants or all products necessary to convert the substrate into product.
  • cyclization refers to the formation of a covalently closed ring by formation of a new bond.
  • reduced and reduction refers to the transfer of one or more electrons to a molecular entity.
  • a compound can be reduced by the addition of hydrogen.
  • a reduced species also can be formed through the gain of electrons.
  • the reverse process in which one or more electrons is removed from a molecular entity is known as "oxidation.”
  • salting refers to the addition of electrolytes to a solution.
  • Salting often alters the distribution ratio of a particular solute or changes the miscibility of two liquids.
  • Shotten-Baumann conditions refers to acylation of alcohols with acyl halides in aqueous alkaline solution.
  • a “derivative” is a compound obtained or produced by modification of another compound of similar structure. Derivatives can be produced by one or more modification steps.
  • sesamol refers to benzo[ ⁇ /][l,3]dioxol-5-ol, otherwise known as 3,4-methylenedioxyphenol.
  • asesamol derivative refers to a compound obtained or produced by modification of sesamol or similar structure. Sesamol derivatives can be produced by one or more modification steps.
  • quenched refers to arresting the course of a chemical or enzymatic reaction by chemical or physical means.
  • protecting groups are often employed. That is, a functional group is temporarily converted into an unreactive form to prevent its interference with transformations to be carried out elsewhere in the molecule. Such temporary functional group modification is known as "protecting" the original group. Subsequent to transformation carried out elsewhere in the molecule, the original unit can be regenerated, i.e., "deprotected," under separate conditions.
  • an alcohol can be protected as a 1,1 -dimethyl ethyl ether by an acid catalyzed reaction of the alcohol with 2-methyl-2-propanol. The resulting ether is inert to some basic, oxidizing, or reducing conditions. The alcohol can be deprotected by removal of the ether group in dilute aqueous acid.
  • Desymmetrization involves the modification of a compound that results in the loss of one or more symmetry elements.
  • Desymmetrization includes the loss of a symmetry element that precludes chirality, such as a mirror plane, center of inversion, or rotational-reflection axis.
  • Desymmetrization can result in the conversion of a prochiral molecular entity into a chiral entity.
  • prochiral refers to a structure that lacks chirality, but can become chiral by addition, removal, or replacement of a substituent.
  • hydrolysis refers to the general rupture of one or more bonds by water molecules.
  • Keevenagel condensation refers to the condensation of aldehydes or ketones with active methylene compounds in the presence of ammonia or amines.
  • ichael addition refers to the base catalyzed addition of methylene compounds to unsaturated systems.
  • EC refers to the Enzyme Commission of the International Union of Biochemistry and Molecular Biology (IUBMB).
  • IUBMB International Union of Biochemistry and Molecular Biology
  • EC numbers such as EC 3.4.21.62, are associated with a recommended name for the respective enzyme. The first number designates the major class, the second number designates the subclass, and the third number designates the sub-subclass. The fourth number indicates the serial number of the enzyme in its sub-subclass.
  • an "esterase” is an enzyme that catalyzes the cleavage of ester bonds. Many esterases show specificity for particular types of esters.
  • a “protease” is an enzyme that catalyzes the hydrolysis of peptide bonds. Many proteases are also capable of cleaving ester bonds. Proteases often show specificity for particular bond arrangements.
  • hydrolase refers to enzymes that catalyze the cleavage of C- O, C-N, C-C and other bonds by reactions involving the addition or removal of water.
  • product refers to a substance that is formed during a chemical or enzymatic reaction.
  • reaction medium refers to the phase in which a chemical or biological reaction or other such transformation takes place.
  • the reaction medium can include solid, liquid, and gaseous phases and mixtures thereof. Chemical and biological reactants and reagents are commonly dissolved or suspended in various liquid compositions to facilitate a reaction or transformation.
  • catalyst is a substance that increases the rate of a reaction.
  • a catalytic substance is a substance that increases the rate of a reaction.
  • biocatalyst refers to a living organism, enzyme, and/or enzyme complex that catalyses a reaction or otherwise facilitates substrate conversion in various chemical reactions.
  • a "buffer solution” is any substance or mixture of compounds in solution that is capable of neutralizing both acids and bases without appreciably changing the original acidity or alkalinity of the solution.
  • Buffer solutions contain mixture(s) of acid and conjugate base at or near the pK a to minimize pH changes caused by an influx of acid or base.
  • Buffer solutions optionally also can contain additional solutes such as salts and other compounds.
  • enzyme “deactivation” occurs when an enzyme is no longer capable of catalysis.
  • a lipase is one of various enzymes that catalyze the hydrolysis of fats, especially triglycerides and phospholipids, into glycerol and fatty acids. Many lipases selectively cleave ester bonds.
  • substrate refers to the chemical entity involved in a reaction that undergoes conversion to a product or products. Enzymes can catalyze the conversion of substrate(s) to product(s).
  • precipitate refers to the act of separating, e.g., a compound or product, from solution or suspension, usually via a chemical or physical change, often resulting in an insoluble oil or amorphous or crystalline solid, and the term also refers to a substance separated from a solution or suspension by chemical or physical change.
  • peak area refers to the area between the peak and the baseline of a chromatogram.
  • co-solvent refers to a mixture of liquids.
  • the co-solvent is a material that is not necessarily an acceptable solvent that is added to a generally small amount of an active solvent to form a mixture that has enhanced solvent power.
  • a polar cosolvent can be added into a mixture of an organic liquid and a compound having pendant ionomeric groups to solubilize the pendant ionomeric groups.
  • Co-solvents can increase solubility of a compound.
  • the use of cosolvents can increase the solubility by several orders of magnitude.
  • Some commonly used cosolvents in pharmaceuticals are propylene glycol, polyethylene glycols, ethanol and sorbitol.
  • the addition of a co-solvent can increase solubility of hydrophobic molecules by reducing the dielectric constant of the solvent.
  • THF tetrahydrofuran
  • the term "article of manufacture” is a product that is made and sold and that includes a container and packaging, and optionally instructions for use of the product.
  • articles of manufacture encompass packaged intermediates as disclosed herein.
  • the articles of manufacture include one or more intermediates as provided herein and an enzyme.
  • the articles of manufacture include one or more intermediates as provided herein and a hydrolase.
  • a “kit” refers to a combination of an intermediate provided herein and another item for a purpose including, but not limited to, synthesis of paroxetine or a related compound. Kits also optionally include instructions for use and/or reagents and glassware and other such items for use with the product.
  • substantially identical to a product means sufficiently similar so that the property of interest is sufficiently unchanged so that the substantially identical product can be used in place of the product.
  • homologous means about greater than 25% nucleic acid sequence identity, generally 25% 40%, 60%, 80%, 90% or 95%.
  • the terms “homology” and “identity” are often used interchangeably.
  • sequences are aligned so that the highest order match is obtained (see, e.g., Computational Molecular Biology, Lesk, A.M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A.M., and Griffin, H.
  • the term "identity" represents a comparison between a test and a reference polypeptide or polynucleotide.
  • a test polypeptide can be defined as any polypeptide that is 90% or more identical to a reference polypeptide.
  • the term “90% identical to” refers to percent identities from 90 to 99.99 relative to the reference polypeptides. Identity at a level of 90% or more is indicative of the fact that, assuming for exemplification purposes a test and reference polynucleotide length of 100 amino acids are compared, no more than 10% (i.e., 10 out of 100) amino acids in the test polypeptide differs from that of the reference polypeptides.
  • Similar comparisons can be made between a test polynucleotide and reference polynucleotides. Such differences can be represented as point mutations randomly distributed over the entire length of an amino acid sequence or they can be clustered in one or more locations of varying length up to the maximum allowable, e.g., in the case of approximately 90% identity, 10/100 amino acid difference. Differences are defined as nucleic acid or amino acid addition, substitutions or deletions. The number of conserved amino acids are determined by standard alignment algorithms programs, and are used with default gap penalties established by each supplier. Substantially homologous nucleic acid molecules would hybridize typically at moderate stringency or at high stringency all along the length of the nucleic acid of interest. Also contemplated are nucleic acid molecules that contain degenerate codons in place of codons in the hybridizing nucleic acid molecule.
  • nucleic acid molecules have nucleotide sequences that are at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% "identical” can be determined using known computer algorithms such as the "FAST A” program, using for example, the default parameters as in Pearson et al, (1988) Proc. Natl. Acad. Sci. USA 85:2444 (other programs include the GCG program package (Devereux et al. , Nucleic Acids Research 12(I):387 (1984)), BLASTP, BLASTN, FASTA (Atschul et al, J Molec Biol 215:403 (1990); Guide to Huge Computers, Martin J.
  • the GAP program defines similarity as the number of aligned symbols ⁇ i.e., nucleotides or amino acids) which are similar, divided by the total number of symbols in the shorter of the two sequences.
  • Default parameters for the GAP program can include: (1) a unary comparison matrix (containing a value of 1 for identities and 0 for non- identities) and the weighted comparison matrix of Gribskov et al, (1986) Nucl. Acids Res. 14:6745, as described by Schwartz and Dayhoff, eds., ATLAS OF PROTEIN SEQUENCE AND STRUCTURE, National Biomedical Research Foundation, pp.
  • Percent homology or identity of proteins and/or nucleic acid molecules can be determined, for example, by comparing sequence information using a GAP computer program (e.g., Needleman et al., (1970) J. MoI. Biol. 48:443, as revised by Smith and Waterman ((1981) Adv. Appl. Math. 2:482). Briefly, the GAP program defines similarity as the number of aligned symbols (i.e., nucleotides or amino acids) which are similar, divided by the total number of symbols in the shorter of the two sequences.
  • a GAP computer program e.g., Needleman et al., (1970) J. MoI. Biol. 48:443, as revised by Smith and Waterman ((1981) Adv. Appl. Math. 2:482).
  • the GAP program defines similarity as the number of aligned symbols (i.e., nucleotides or amino acids) which are similar, divided by the total number of symbols in the shorter of the two sequences.
  • Default parameters for the GAP program can include: (1) a unary comparison matrix (containing a value of 1 for identities and 0 for non-identities) and the weighted comparison matrix of Gribskov et al., (1986) Nucl. Acids Res. 14:6745, as described by Schwartz and Dayhoff, eds., ATLAS OF PROTEIN SEQUENCE AND STRUCTURE, National Biomedical Research Foundation, pp. 353-358 (1979); (2) a penalty of 3.0 for each gap and an additional 0.10 penalty for each symbol in each gap; and (3) no penalty for end gaps.
  • heterologous or foreign DNA and RNA are used interchangeably and refer to DNA or RNA that does not occur naturally as part of the genome in which it is present or which is found in a location or locations in the genome that differ from that in which it occurs in nature.
  • Heterologous nucleic acid is generally not endogenous to the cell into which it is introduced, but has been obtained from another cell or prepared synthetically. Generally, although not necessarily, such nucleic acid encodes RNA and proteins that are not normally produced by the cell in which it is expressed. Any DNA or RNA that one of skill in the art would recognize or consider as heterologous or foreign to the cell in which it is expressed is herein encompassed by heterologous DNA.
  • Heterologous DNA and RNA also can encode RNA or proteins that mediate or alter expression of endogenous DNA by affecting transcription, translation, or other regulatable biochemical processes.
  • amino acids which occur in the various amino acid sequences appearing herein, are identified of their well-known, three-letter or one- letter abbreviations.
  • conservative amino acid substitutions can be made in any of hydrolases provided that the resulting protein exhibits hydrolase activity.
  • Conservative amino acid substitutions, such as those set forth in Table 1 are those that do not eliminate hydrolase activity. Suitable conservative substitutions of amino acids are known to those of skill in this art and can be made generally without altering the biological activity of the resulting molecule.
  • Contemplated amino acid substitutions include any amino acid substitution such that the enzyme retains its catalytic activity.
  • the term "monitoring” refers to observing an effect or absence of any effect. In certain embodiments, one monitors a reaction after addition of a reactant or change in reaction conditions, such as temperature or pressure. Examples of effects that can be monitored include, but are not limited to, changes in evolution of gas, the appearance of a reaction product or a disappearance of a substrate or reactant.
  • the term “contacting” refers to bringing two or more materials into close enough proximity that they can interact. In certain embodiments, contacting can be accomplished in a vessel such as, e.g., a test tube, flask, petri dish or mixing tank. In certain embodiments, contacting can be performed in the presence of additional materials.
  • the term "subject" is an animal, typically a mammal, including human.
  • the term “patient” includes human and animal subjects.
  • carrier refers to a compound that facilitates the incorporation of another compound into cells or tissues.
  • DMSO dimethyl sulfoxide
  • a pharmaceutical composition refers to a chemical compound or composition capable of inducing a desired therapeutic effect in a subject.
  • a pharmaceutical composition contains an active agent, which is the agent that induces the desired therapeutic effect.
  • the pharmaceutical composition can contain a prodrug of the compounds provided herein.
  • a pharmaceutical composition contains inactive ingredients, such as, for example, carriers and excipients.
  • therapeutically effective amount refers to an amount of a pharmaceutical composition sufficient to achieve a desired therapeutic effect.
  • pharmaceutically acceptable refers to a formulation of a compound that does not significantly abrogate the biological activity, a pharmacological activity and/or other properties of the compound when the formulated compound is administered to a subject. In certain embodiments, a pharmaceutically acceptable formulation does not cause significant irritation to a subject.
  • pharmaceutically acceptable derivatives of a compound include, but are not limited to, salts, esters, enol ethers, enol esters, acetals, ketals, orthoesters, hemiacetals, hemiketals, acids, bases, solvates, hydrates or prodrugs thereof.
  • Such derivatives can be readily prepared by those of skill in this art using known methods for such derivatization.
  • the compounds produced can be administered to animals or humans without substantial toxic effects and either are pharmaceutically active or are prodrugs.
  • Pharmaceutically acceptable salts include, but are not limited to, amine salts, such as but not limited to chloroprocaine, choline, N,N'-dibenzyl-ethylenediamine, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N- methylglucamine, procaine, N-benzyl-phenethylamine, 1 -para-chloro-benzyl-2- pyrrolidin-l'-yl-methylbenzimidazole, diethylamine and other alkylamines, piperazine and tris(hydroxyrnethyl)-aminomethane; alkali metal salts, such as but not limited to lithium, potassium and sodium; alkali earth metal salts, such as but not limited to barium
  • esters include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl and heterocyclyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids and boronic acids.
  • Pharmaceutically acceptable solvates and hydrates are complexes of a compound with one or more solvent or water molecules, or 1 to about 100, or 1 to about 10, or one to about 2, 3 or 4, solvent or water molecules.
  • membrane reactor refers to any reaction vessel in which the catalyst, such as an enzyme, is enclosed in a reactor, either in solution or attached to a solid support, while low-molecular substances within the reactor are able to leave the reactor.
  • the membrane of the reactor can be integrated directly into the reaction chamber.
  • the membrane is incorporated outside the reactor chamber in a separate filtration module, with the reaction solution flowing continuously or intermittently through the filtration module, and with the retentate being recirculated into the reactor.
  • paroxetine refers to 3-((benzo[d][l,3]dioxol-5- yloxy)methyl)-4-(4-fluorophenyl)piperidine or a pharmaceutically acceptable salt thereof.
  • "paroxetine” refers to the (3S,4R) configuration of 3-((benzo[d][l,3]dioxol-5-yloxy)methyl)-4-(4-fluorophenyl)piperidine or a pharmaceutically acceptable salt thereof.
  • paroxetine there are several synthetic methods known in the art for preparing paroxetine (see e.g., U.S. 3,912,743; U.S. 4,007,196; U.S. 4,721,723; U.S. 4,902,801 ; EP 0223403; Lie et al, Tet. Asym. 12, 419, 2001; and Gonzalo et al, J. Org. Chem. 68, 3333, 2003).
  • Successful preparation of active paroxetine requires an optically active product of the 3S,4R configuration.
  • a focus of the prior art methods is the preferential isolation of trans-3,4-disubstituted piperidine derivatives. For example, U.S. Pat. No.
  • 4,007,196 produces the trans orientation by base-catalyzed epimerization.
  • a symmetrical 4-aryl substituted glutarimide was desymmetrized by enolization with a chiral base. This process can afford a 3,4- disubstituted glutarimide derivative of the trans orientation by subsequent treatment with an electrophile (see Greenhalgh et al, Synlett 12, 2074, 2002; and Gill et al, Tetrahedron 59, 9213, 2003).
  • Other methods resolve trans isomers via biocatalysis of piperidine-ester substrates (see WO 01/029032; WO 93/22284; de Gonzalo et al, J. Org. Chem.
  • Scheme I demonstrates a multi-step route to prepare optically pure paroxetine (see WO 02/32870; WO 98/02556; WO 97/44320; WO 01/85688; and WO 94/03428).
  • the synthetic pathway starts with the natural product arecoline, which is reacted with p-fluorophenyl grignard using the method originally described by Plati et al. (J. Org. Chem., 1957, 22, 261). This produces the disubstituted piperidine (see also WO 01/29032).
  • the trans orientation can be obtained as the predominant form via base- catalyzed epimerization.
  • the menthol ester was originally resolved (see U.S.
  • diastereomeric salts can be difficult to selectively recrystalize (see Liu et al., Tetrahedron Asymmetry 2001, 12, 419-426).
  • the use of chiral auxiliary groups also can add additional synthetic steps.
  • Selective enzymatic resolution has been reported ⁇ e.g., WO 01/29032). Such methods, however, are not a significant improvement over chemical resolution since both methods waste half of the synthesized material.
  • a desymmetrization process of a symmetrical imide containing two ester functionalities is achieved by selective enzyme hydrolysis of a single ester group.
  • hydrolases catalyze the hydrolysis of ester bonds, including, but not limited to, various proteases and esterases.
  • the methods provided herein can use an esterase alone or in combination with other esterases.
  • Scheme II a chiral imide ester of 3S,4R configuration is obtained by this process. This approach allows for a theoretical 100% yield of chirality installation.
  • the desymmetrization step described herein proceeds without loss of significant starting material.
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, C 1 -C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Cj-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted; and
  • R' and R" each independently is selected from among hydrogen, C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, CpC 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among C)-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, hetero
  • substituents can be selected from among a subset of the listed alternatives.
  • hydrolases Any hydrolase, alone or in combination with other hydrolases, can be used. If needed, hydrolases, libraries thereof and/or libraries of modified hydrolases can be screened to identify more suitable or suitable hydrolases for the methods provided herein.
  • Hydrolases are enzymes that catalyze the hydrolysis of a chemical bond. Hydrolases comprise the third major class of enzymes, EC 3, in the Enzyme Commission of the International Union of Biochemistry and Molecular Biology (IUBMB) numbering classification system. Hydrolases are further classified into several subclasses and sub-subclasses. Each hydrolase receives a serial number within the corresponding sub-subclass. Bonds that can be hydrolyzed by these enzymes include, but are not limited to, ester bonds, bonds to sugars, ether bonds, peptide bonds, carbon-nitrogen bonds, acid anhydrides, carbon-carbon bonds, halide bonds, phosphorus-nitrogen bonds, sulfur-nitrogen bonds, carbon-phosphorus bonds, sulfur- sulfur bonds and carbon-sulfur bonds.
  • hydrolases can hydrolyze esters, including, but not limited to, various proteases, and those enzymes classified as esterases, including lipases. Any enzyme that can hydrolyze the ester bond as shown in Scheme II can be used in the methods described herein.
  • the hydrolase is an esterase, a lipase, or a protease or combinations thereof.
  • the hydrolase can be of mammalian, including human, origin, or can be of non-mammalian origin, including but not limited to, plant, bacterial, viral, yeast and fungal origin.
  • the hydrolase used in the methods provided herein to hydrolyze the ester bond can be a wild-type protein or a variant thereof.
  • hydrolase variant is used that contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more mutations, such as amino acid substitutions, deletions, additions, or combinations thereof.
  • hydrolase variants can be generated by directed evolution such that the substrate specificity and enzymatic activity of the variants are optimized for the purposes herein ⁇ see, e.g., Bornscheuer et al., Curr. Opin. Biotech. 7:2169-2173 (1999)).
  • the properties of a candidate hydrolase, such as substrate specificity and enzymatic activity can be assessed using any method known in the art, as described below.
  • high-throughput screening of multiple hydrolases is performed ⁇ e.g., see Yazbeck et al., Adv. Synth. Catal.
  • Esterases (classified as EC 3.1 in the EC number classification of enzymes) are enzymes that hydrolyze esters into an acid and an alcohol in a chemical reaction with water. This is mediated through nucleophilic attack of the active serine in the esterase on the carbonyl of the substrate in a charge-relay system with two other amino acid residues in the esterase. Together, the three amino acid residues are called the catalytic triad. Esterases display broad substrate specificity and also can exhibit enantioselectivity. Many esterases have been described in the art that hydrolyze different substrates.
  • esterase enzymes cleaving carboxyl esters have been identified in leukocytes; phosphatases, such as alkaline and acidic phosphatases, hydrolyse phosphoric acid esters; glycosidases, such as galactosidases, glucosidases, mannosidases and amylases can cleave glycosidic bonds.
  • phosphatases such as alkaline and acidic phosphatases, hydrolyse phosphoric acid esters
  • glycosidases such as galactosidases, glucosidases, mannosidases and amylases can cleave glycosidic bonds.
  • esterases include, but are not limited to, lipases, acetylesterases, thiolester hydrolases, phosphoric monoester hydrolases, phosphoric diester hydrolases, triphosphoric monoester hydrolases, sulfuric ester hydrolases, diphosphoric monoester hydrolases, phosphoric triester hydrolases, exonucleases and endonucleases.
  • lipases acetylesterases, thiolester hydrolases, phosphoric monoester hydrolases, phosphoric diester hydrolases, triphosphoric monoester hydrolases, sulfuric ester hydrolases, diphosphoric monoester hydrolases, phosphoric triester hydrolases, exonucleases and endonucleases.
  • lipases acetylesterases, thiolester hydrolases, phosphoric monoester hydrolases, phosphoric diester hydrolases, triphosphoric monoester hydrolases, sulfuric ester
  • Lipases also known as triacylglycerol ester hydrolases, (E. C.3.1.1.3) are a subclass of esterases that belong to the al ⁇ hydrolase super family. They are ubiquitous enzymes that can be generally divided into the following four groups of their specificity in hydrolysis reaction: substrate specific lipases, regio-selective lipases, fatty acid specific lipases, and stereo-specific lipases. Lipases can be obtained from a variety of sources including, but not limited to, plants, animals, yeast and bacteria.
  • Exemplary lipases include, but are not limited to, lipases from Acinetobacter calcoaceticus, Acinetobacter sp., Alcaligenes sp., Aspergillus carneus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Bacillus sp.
  • Candida rugosa One of skill in the art can screen any lipase for substrate specificity and enzymatic activity to determine its suitability for use in the methods provided herein for the production of paroxetine or related compounds. c. Proteases
  • Esters also can be hydrolyzed by proteases, including, but not limited to, serine proteases and cysteine proteases.
  • proteases including, but not limited to, serine proteases and cysteine proteases.
  • the activity of proteases in the serine protease family is dependent on a set of amino acid residues that form the active site. One of the residues is always a serine, hence their designation as serine proteases.
  • serine proteases also can hydrolyze ester bonds. They can be obtained from a variety of sources including, but not limited to, animals (including humans), plants, yeast and bacteria.
  • Exemplary eukaryotic serine proteases include, but are not limited to, acrosin; blood coagulation factors VII, IX, X, XI and XII, thrombin, plasminogen, and protein C; cathepsin G; chymotrypsins; complement components CIr, CIs, C2, and complement factors B, D and I; complement-activating component of RA-reactive factor; cytotoxic cell proteases (granzymes A to H); duodenase I; elastases 1, 2, 3 A, 3B (protease E), leukocyte (medullasin); enterokinase (enteropeptidase); hepatocyte growth factor activator; hepsin; glandular (tissue) kallikreins (including EGF-binding protein types A, B, and C, NGF - ⁇ chain, ⁇ -renin, prostate specific antigen (PSA) and tonin); plasma kallikrein; mast
  • Exemplary prokaryotic serine protease include, but are not limited to, subtilisins from Bacillus sp.; alkaline elastase YaB from Bacillus sp.; alkaline serine exoprotease A from Vibrio alginolyticus; aqualysin I from Thermus aquaticus; AspA from Aeromonas salmonicida; bacillopeptidase F (esterase) from Bacillus subtilis; C5A peptidase from Streptococcus pyogenes; cell envelope-located proteases PI, PII, and PIII from Lactococcus lactis; extracellular serine protease from Serratia marcescens; extracellular protease from Xanthomonas campestris; intracellular serine protease (ISP) from various Bacillus; minor extracellular serine protease epr from Bacillus subtilis; minor extracellular serine protease
  • alkaline extracellular protease (AEP) from Yarrowia lipolytica; alkaline proteinase from Cephalosporium acremonium; cerevisin (vacuolar protease B) from yeast; cuticle-degrading protease (prl) from Metarhizium anisopliae.; KEX-I protease from Kluyveromyces lactis.
  • AEP alkaline extracellular protease
  • Cephalosporium acremonium alkaline proteinase from Cephalosporium acremonium
  • cerevisin (vacuolar protease B) from yeast
  • cuticle-degrading protease (prl) from Metarhizium anisopliae.
  • KEX-I protease from Kluyveromyces lactis.
  • kexin from yeast
  • oryzin alkaline proteinase from Aspergillus
  • proteinase K from Tritirachium album
  • proteinase R from Tritirachium album
  • proteinase T from Tritirachium album
  • subtilisin-like protease III from yeast
  • thermomycolin from Malbranchea sulfurea.
  • cysteine proteases Members of the class of cysteine proteases have a common catalytic mechanism that involves a cysteine amino acid residue in the active site of the protease.
  • Exemplary cysteine proteases that can be used in the methods herein include, but are not limited to, vertebrate lysosomal cathepsins B, H, L, and S; vertebrate lysosomal dipeptidyl peptidase I (also known as cathepsin C); vertebrate calpains; mammalian cathepsin K, which seems involved in osteoclastic bone resorption; human cathepsin O; bleomycin hydrolase; barley aleurain; EP-B 1/B4; kidney bean EP-Cl ; rice bean SH-EP; kiwi fruit actinidin; papaya latex papain; chymopapain; caricain; proteinase IV; pea turgor-responsive protein 15 A; pineapple stem bromel
  • hydrolases useful for the methods herein Any method known in the art to screen enzymes for substrate specificity and enzymatic activity can be used to identify hydrolases useful for the methods provided herein ⁇ see e.g., Gupta et al, (2003) Biotechnol. Appl. Biochem. 37:63-71 ; Yazbeck et al, (2003) Adv. Synth. Catal. 345:524-532; and Kim et al, (2006) Prot. Exp. Purif. 45:315-323).
  • high throughput methods are employed to screen multiple enzymes for their ability to hydrolyze the ester bond. Such methods are typically performed in, for example, 96-well microtiter plates, such that multiple hydrolases ⁇ e.g., a hydrolase library) can be simultaneously screened for hydrolysis of a chosen substrate.
  • the hydrolases are obtained from commercial sources, such as those provided in Table 2.
  • the hydrolases are specifically engineered for this purpose, such as through directed evolution methods (see Bornscheuer et al, (1999) Curr. Opin. Biotech. 7:2169-2173).
  • the amino acid sequences of exemplary hydrolases that can be used in the methods herein are set forth in SEQ ID NOS: 1-187.
  • EC number, enzyme names, species and the sequence identifier (SEQ ID NO) of such enzymes are provided in Table 3.
  • exemplary carboxylesterases, including pig liver esterase are set forth in SEQ ID NOS: 1-3 and 28-113;
  • the screening reactions are typically carried out in volumes of about 100 ⁇ l of a suitable buffer, such as 0.1 M potassium phosphate buffer, containing 1 mg/ml of substrate, 10 mg/ml of enzyme and 10% organic solvent.
  • a suitable buffer such as 0.1 M potassium phosphate buffer
  • the pH of the reaction is generally maintained at between 7.0 and 7.4.
  • water immiscible solvents such as methyl tert-buty ⁇ ether (MTBE), ethyl acetate, dichloro-methane, toluene or diisopropyl ether (DIPE) can be used for lipase screening, resulting in a biphasic system.
  • MTBE methyl tert-buty ⁇ ether
  • DIPE diisopropyl ether
  • Proteases and esterases can be screened in water miscible solvents, such as acetonitrile, methanol, acetone or ethanol. Following incubation at an appropriate temperature, typically ranging from O 0 C to 6O 0 C, the samples are analyzed to assess hydrolysis.
  • a variety of methods can be used in the analysis of the sample, including, but not limited to, high performance liquid chromatography (HPLC), capillary electrophoresis (CE), gas chromatography (GC), UV spectrophotometry, thin layer chromatography (TLC) and liquid chromatography coupled with mass spectrometry (LC-MC). e.
  • Suitable organic solvents that can be used for this purpose include, but are not limited to, hexane, cyclohexane, ethyl acetate, 1 -hexanol, chloroform, dichloroethane, dichloromethane, toluene, tert-pentyl alcohol, methyl isobutyl ketone (MIBK), methyl tert-butyl ether (MTBE) and di-isopropyl ether (DIPE).
  • the reaction is carried out in an aqueous buffer.
  • Suitable aqueous phases include, but are not limited to, buffers such as glutamic acid- glutamate, phosphoric acid-phosphate, acetic acid-acetate and citric acid-citrate buffers.
  • buffers such as glutamic acid- glutamate, phosphoric acid-phosphate, acetic acid-acetate and citric acid-citrate buffers.
  • 0.1 M potassium phosphate buffer can be used in the methods herein.
  • a suitable pH for the reaction can be determined by one of skill in the art, taking into account the activity and stability of the hydrolase. A pH range from 3 to 11 is contemplated for the methods herein.
  • a neutral environment is maintained, such that the pH of the reaction is at or about 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6 or 7.7.
  • the temperature during the reaction can be between O 0 C to 6O 0 C. In some embodiments, an ambient temperature is maintained throughout the reaction. In other embodiments,
  • the amount of hydrolase utilized is determined by the activity of the enzyme.
  • An IU International Unit designates that amount of an enzyme preparation that catalyzes the formation of one micromole of product per minute. Such determinations can be made using methods well known in the art. For the methods herein, typically 10 to 10,000 IU of hydrolase is added to the reaction for every gram of substrate. The mixture is then typically agitated throughout the reaction. The reaction can be monitored by, for example, gas chromatography (GC), high performance liquid chromatography (HPLC) or thin layer chromatography (TLC) to determine the point of completion and the optical purity of the product. The resulting compound can be isolated by extraction, evaporation, or other suitable separation methods.
  • GC gas chromatography
  • HPLC high performance liquid chromatography
  • TLC thin layer chromatography
  • the reaction is monitored by GC and the product is purified by extraction.
  • the reaction mixture optionally is saturated with sodium chloride and repeatedly extracted with ethyl acetate until complete recovery of the product.
  • the organic layers are then dried over anhydrous sodium sulfate and filtered and concentrated.
  • D. Intermediates In certain embodiments, the compounds provided herein have a structure of
  • R is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, CpC 8 haloalkyl, C 1 -C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", CpC 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted;
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, C 1 -C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted; or a pharmaceutically acceptable salt of the compound of formula I, with the proviso that if R 2 is methyl and X is 4-fluoro, then R 1 is not methyl or benzyl.
  • substituents can be selected from among a subset of the listed alternatives.
  • R 1 is selected from among hydrogen, OR',
  • R'R C 1 -C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, Cj-C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 -C 9 )alkyl having one to eight heteroatoms selected from among O, N, S and P, C 3 -C 9 aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, C 1 -C 8 heteroalkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl, wherein the alkyl, alkenyl, alkynyl, halo
  • R 1 is selected from among hydrogen, OR', NR 'R", C 1 - C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cyclo(C 3 - C 9 )alkyl, heterocyclo(C 3 -C 9 )alkyl having one to eight heteroatoms selected from among O, N, S and P, C 3 -Cg aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, C 1 -C 8 heteroalkyl, C 3 -Cg aryl and C 3 -Cg heteroaryl, wherein the al
  • R 1 is selected from among C 1 -C 8 alkyl, C 2 -C 8 alkenyl,
  • R 1 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci -C 8 haloalkyl, Q-C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - Cg)-alkyl having one to eight heteroatoms selected from among O, N, S and P, C 3 -C 9 aryl and C 3 -Cg heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci -6 - alkoxy, Ci -6 -alky
  • R 1 is selected from among C 2 -C 8 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 8 haloalkyl, C 2 -C 8 heteroalkyl, cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 - C 8 )-alkyl having one to six heteroatoms selected from among O, N, S and P, C 4 -C 8 aryl and C 4 -Cg heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 - alkoxy,
  • R 1 is selected from among C 2 -C 8 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 8 haloalkyl, C 2 -C 8 heteroalkyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl and heteroalkyl, are optionally substituted with a substituent selected from among halo, hydroxy, amino,- cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 - alkyl, Ci-C 6 -haloalkyl, Ci-C 6 -hydroxy-alkyl, Ci-C 6 -aminoalkyl, Ci-C 6 -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl and trifluoromethyl.
  • R 1 is selected from among cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 -C 8 )-alkyl having one to six heteroatoms selected from among O, N, S and P, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C 6 -haloalkyl, Ci-C 6 -hydroxy-alkyl, Ci-C 6 -aminoalkyl, Ci-C 6 -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsul
  • R 1 is selected from among hydrogen, Ci-C 6 -haloalkyl, Ci-C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 arylalkyl, Ci-C 6 -hydroxy-alkyl and hydroxy.
  • R 1 is selected from among hydrogen, trifluoro(Ci- C 4 )-alkyl, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C r C 4 arylalkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl.
  • R 1 is selected from among hydrogen, fluoro(Ci-C 4 )-alkyl, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl. In other embodiments, R 1 is selected from among hydrogen, C 2 -C 6 alkyl, CpC 4 arylalkyl, C 3 -C 6 alkenyl and benzyl. In other embodiments, R 1 is selected from among hydrogen, C 1 -C 4 haloalkyl, and C 2 -C 6 alkyl. In other embodiments, R 1 is hydrogen. In other embodiments, R 1 is Ci-C 6 haloalkyl. In other embodiments, R 1 is Ci-C 6 alkyl. In other embodiments, R 1 is methyl. In other embodiments, R 1 is hydrogen. In other embodiments, R 1 is benzyl.
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - C 9 )alkyl, C 3 -Cg aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among Ci-C 8 alkyl, cyclo(C 3 -C 9 )alkyl, C 1 -C 8 heteroalkyl, C 3 -C 9 aryl, C 3 -C 9 heteroaryl, non-aromatic
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Cj-C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - C 9 )alkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C ⁇ -haloalkyl,
  • R 2 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl, C 2 -C 6 heteroalkyl, cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 - C 8 )alkyl, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -alkoxy, C]-C 4 -alkyl, Ci-C4-halo
  • R 2 is selected from among C 1 -C 4 alkyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl, C4-C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkynyl, haloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -alkoxy, Ci-C 4 -alkyl, Ci-C 4 - haloalkyl, Ci-C4-hydroxy-alkyl, Ci-C 4 -aminoalkyl, Ci-C 4 -alkylamino, Ci-C 4 - alkylsulfenyl, C 1 -C 4 -alkylsulf ⁇ nyl, Ci-C
  • R 2 is selected from among C 1 -C 4 alkyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl and benzyl, wherein the alkyl, alkynyl, haloalkyl and benzyl are optionally substituted with a substituent selected from among halo, Ci-C 4 -alkyl, Ci- C 4 -haloalkyl, Ci-C 4 -alkylsulfenyl, Ci-C 4 -alkylsulfinyl, Ci-C 4 -alkylsulfonyl, amino, cyano, nitro, alkylamido, acyl and Ci-C 4 -alkoxy.
  • R 2 is C 1 -C4 alkyl. In some embodiments, R 2 is C 2 -C 6 alkynyl. In some embodiments, R 2 is benzyl. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is Ci-C 6 heteroalkyl. In some embodiments, R is Cj-C 6 alkoxy. In some embodiments, R 2 is -CH 2 (CH 2 )o- 6 ⁇ H.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR 1 R", CpC 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 -C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalky
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Cj-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 -C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, wherein the al
  • X is selected from among OR', SR', SOR', SO 2 R', CO 2 R' and NR'R", wherein R' and R" each independently is selected from among hydrogen, C r C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 - C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P and C 3 -C 9 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl
  • X is selected from among OR', SR', SOR', SO 2 R' and CO 2 R', wherein R' is selected from among hydrogen, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 - C 6 alkynyl, Ci-C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 8 heteroaryl having one to seven heteroatoms selected from among O, N, S and P and C 4 -C 8 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -alk
  • X is selected from among OR', SR', SOR', SO 2 R' and CO 2 R', wherein R' is selected from among hydrogen, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 - C 6 alkynyl, CpC 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 8 heteroaryl having one to seven heteroatoms selected from among O, N, S and P and C 4 -C 8 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl and trifluoromethyl.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci -C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci- C 6 -alkyl, Ci-C
  • X is selected from among F, Cl, Br, I, CN, NO 2 , C 2 -C 4 alkyl, C 2 -C 4 haloalkyl and Ci-C 4 heteroalkyl, wherein the alkyl, haloalkyl and heteroalkyl, are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C 6 - haloalkyl, Ci-C ⁇ -hydroxy-alkyl, C]-C 6 -aminoalkyl, Cj-C ⁇ -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl and trifluoromethyl.
  • X is selected from among F, Cl, Br, I, CN and NO 2 . In some embodiments, X is selected from among F, Cl, Br and I. In some embodiments, X is F. In some embodiments, X is 4-fluoro. In some embodiments, X is hydrogen.
  • X is trifluoroalkyl. In some embodiments, X is hydroxyl. In some embodiments, X is CpC 4 alkyl. In some embodiments, X is Ci-C 4 alkoxy.
  • R 1 is selected from among hydrogen, Ci-C 8 alkyl, C 2 -
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl; and
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Cj-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 - C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N,
  • R 1 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl,
  • R 2 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 haloalkyl, CpC 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 7 cycloalkyl, C 4 -C 6 heterocycloalkyl having one to five heteroatoms selected from among O, N, S and P, phenyl and benzyl; and
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, C)-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl.
  • the compound is selected from among: meso-(3R,4s,5S)-diethyl 4-(4-fluorophenyl)-l-methyl-2,6-dioxopiperidine- 3,5-dicarboxylate; meso-(3R,4s,5S)-diisopropyl 4-(4-fluorophenyl)- 1 -methyl-2,6- dioxopiperidine-3,5-dicarboxylate; /we.sO-(3R,4s,5S)-dibenzyl 4-(4-fluorophenyl)-l-methyl-2,6-dioxopiperidine-
  • the compounds provided herein have a structure of Formula I ⁇ :
  • R 1 is selected from among hydrogen, OR', NR'R", C r C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, hetero
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, CpC 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted; and
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, CpC 8 haloalkyl, Ci-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted; or a pharmaceutically acceptable salt of Formula I A .
  • substituents can be selected from among a subset of the listed alternatives.
  • R 1 is selected from among hydrogen
  • R 1 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, CpC 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - C 9 )-alkyl having one to eight heteroatoms selected from among O, N, S and P, C 3 -Cg aryl and C 3 -Cg heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci -6 - alkoxy, Cj -6 - alkoxy
  • R 1 is selected from among C 2 -C 8 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 8 haloalkyl, C 2 -C 8 heteroalkyl, cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 - C 8 )-alkyl having one to six heteroatoms selected from among O, N, S and P, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 - alkoxy,
  • R 1 is selected from among C 2 -C 8 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 8 haloalkyl, C 2 -C 8 heteroalkyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl and heteroalkyl, are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, C 1 -C 6 - alkyl, Ci-C 6 -haloalkyl, Ci-C 6 -hydroxy-alkyl, Ci-C 6 -aminoalkyl, Ci-C ⁇ -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl and trifluoromethyl.
  • R 1 is selected from among cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 -C 8 )-alkyl having one to six heteroatoms selected from among O, N, S and P, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-Cg-alkoxy, Ci-C 6 -alkyl, Ci-C 6 -haloalkyl, Ci-C 6 -hydroxy-alkyl, Ci-C 6 -aminoalkyl, C ! -C 6 -alkylamino, alkylsulfenyl, alkylsulfinyl, alkyl
  • R 1 is selected from among hydrogen, Ci-C 6 -haloalkyl, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C r C 6 arylalkyl, Ci-C 6 -hydroxy-alkyl and hydroxy.
  • R 1 is selected from among hydrogen, trifluoro(Ci- C 4 )-alkyl, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 4 arylalkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl.
  • R 1 is selected from among hydrogen, fluoro(Ci-C 4 )-alkyl, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl. In other embodiments, R 1 is selected from among hydrogen, C 2 -C 6 alkyl, Ci-C 4 arylalkyl, C 3 -C 6 alkenyl and benzyl. In other embodiments, R 1 is selected from among hydrogen, Ci-C 4 haloalkyl, and C 2 -C 6 alkyl. In other embodiments, R 1 is hydrogen. In other embodiments, R 1 is CpC 6 haloalkyl. In other embodiments, R 1 is C 1 -C 6 alkyl. In other embodiments, R 1 is methyl. In other embodiments, R 1 is hydrogen. In other embodiments, R 1 is benzyl.
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Cj-C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - C 9 )alkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among C 1 -C 8 alkyl, cyclo(C 3 -C 9 )alkyl, Ci-C 8 heteroalkyl, C 3 -C9 aryl, C 3 -C 9 heteroaryl, non-aro
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C r C 8 haloalkyl, C r C 8 heteroalkyl, cyclo(C 3 -C 9 )alkyl, heterocyclo(C 3 - C 9 )alkyl, C 3 -C 9 aryl and C 3 -C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, d-C 6 -alkyl, Ci-C 6 -haloalkyl
  • R 2 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl, C 2 -C 6 heteroalkyl, cyclo(C 4 -C 8 )alkyl, heterocyclo(C 4 - C 8 )alkyl, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -alkoxy, Ci-C 4 -alkyl, Ci-C 4 -halo
  • R 2 is selected from among Ci-C 4 alkyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl, C 4 -C 8 aryl and C 4 -C 8 heteroaryl having one to six heteroatoms selected from among O, N, S and P, wherein the alkyl, alkynyl, haloalkyl, aryl and heteroaryl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -alkoxy, Cj-C 4 -alkyl, C 1 -C 4 - haloalkyl, Ci-C 4 -hydroxy-alkyl, Ci-C 4 -aminoalkyl, C !
  • R 2 is selected from among Ci-C 4 alkyl, C 2 -C 6 alkynyl, C 2 -C 6 haloalkyl and benzyl, wherein the alkyl, alkynyl, haloalkyl and benzyl are optionally substituted with a substituent selected from among halo, Ci-C 4 -alkyl, C 1 - C 4 -haloalkyl, Ci-C 4 -alkylsulfenyl, Ci-C 4 -alkylsulfinyl, Ci-C 4 -alkylsulfonyl, amino, cyano, nitro, alkylamido, acyl and Ci-C 4 -alkoxy.
  • R 2 is Ci-C 4 alkyl. In some embodiments, R 2 is C 2 -C 6 alkynyl. In some embodiments, R 2 is benzyl. In some embodiments, R 2 is methyl. In some embodiments, R 2 is ethyl. In some embodiments, R 2 is Ci-C 6 heteroalkyl. In some embodiments, R 2 is Ci-C 6 alkoxy. In some embodiments, R 2 is -CH 2 (CH 2 ) 0-6 OH.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, CpC 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 -C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalky
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 -C 4 heteroalkenyl having one to three heteroatoms selected from among O, N, S and P and C 2 -C 4 heteroalkynyl having one to three heteroatoms selected from among O, N, S and P, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalky
  • X is selected from among OR', SR', SOR', SO 2 R', CO 2 R' and NR'R", wherein R' and R" each independently is selected from among hydrogen, C r C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C r C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 - C 9 heteroaryl having one to eight heteroatoms selected from among O, N, S and P and C 3 -C 9 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acy
  • X is selected from among OR', SR', SOR', SO 2 R' and CO 2 R', wherein R' is selected from among hydrogen, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 - C 6 alkynyl, C 1 -C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 8 heteroaryl having one to seven heteroatoms selected from among O, N, S and P and C 4 -C 8 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 4 -
  • X is selected from among OR', SR', SOR', SO 2 R' and CO 2 R', wherein R' is selected from among hydrogen, C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 - C 6 alkynyl, Ci-C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 8 heteroaryl having one to seven heteroatoms selected from among O, N, S and P and C 4 -C 8 aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl and trifluoromethyl.
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Ci-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl and heteroalkynyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Cj-C ⁇ -alkoxy, Ci- C 6 -alkyl, Ci-
  • X is selected from among F, Cl, Br, I, CN, NO 2 , C 2 -C 4 alkyl, C 2 -C 4 haloalkyl and Ci-C 4 heteroalkyl, wherein the alkyl, haloalkyl and heteroalkyl, are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C ⁇ -alkoxy, Ci-C 6 -alkyl, Ci-C 6 - haloalkyl, Ci-C 6 -hydroxy-alkyl, Ci-C 6 -aminoalkyl, Ci-C 6 -alkylamino, alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl and trifluoromethyl.
  • X is selected from among F, Cl, Br, I, CN and NO 2 . In some embodiments, X is selected from among F, Cl, Br and I. In some embodiments, X is F. In some embodiments, X is 4-fluoro. In some embodiments, X is hydrogen. In some embodiments, X is trifluoroalkyl. In some embodiments, X is hydroxyl. In some embodiments, X is C 1 -C 4 alkyl. In some embodiments, X is Ci-C 4 alkoxy.
  • R 1 is selected from among hydrogen, C 1 -C 8 alkyl, C 2 - C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Cj-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl having one to seven heteroatoms selected from among O, N, S and P, C 3 -C 8 cycloalkyl, C 3 -C 8 heterocycloalkyl having one to seven heteroatoms selected from among O, N, S and P, phenyl and benzyl; and X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , C r C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl having one to three heteroatoms selected from among O, N, S and P, C 2 - C
  • R 1 is selected from among C 2 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 haloalkyl, Ci-C 6 heteroalkyl having one to five heteroatoms selected from among O, N, S and P, C 4 -C 7 cycloalkyl, C 4 -C 7 heterocycloalkyl having one to five heteroatoms selected from among O, N, S and P, phenyl and benzyl, wherein the alkyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, phenyl and benzyl are optionally substituted with a substituent selected from among halo, hydroxy, amino, cyano, nitro, alkylamido, acyl, Ci-C 6 -alkoxy, Ci-C 6 -alkyl, Ci-C 6 - haloalkyl, Ci-C 6 -hydroxy
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , Cj-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 -C 4 haloalkyl, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl.
  • Scheme IV describes methods to prepare compounds of structures 5 and 6 from compounds of structure 1.
  • R 1 is selected from among hydrogen, OR', NRTl", Cj-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Cj-C 8 haloalkyl, Cj-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 and R 3 each independently is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, CpCg heteroalkyl, cycloalkyl, hetero- cycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl,
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Cj-C 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • compounds of structure 1 are commercially available and known in the art.
  • compounds of structure 1 include, but are not limited to, 4- fluorobenzaldehyde (available from Sigma-Aldrich Co., St. Louis, MO, Cat. No. 128376), 4-chlorobenzaldehyde (Sigma-Aldrich Cat. No. 112216), 4- bromobenzaldehyde (Sigma-Aldrich Cat. No. B57400), or 4-formylbenzonitrile (Sigma-Aldrich Cat. No.C89609).
  • the imide of structure 4 can be prepared by reacting compounds of structure 3 with an amine, such as methylamine, in the presence Of SOCl 2 , 1,1'- carbonyldiimidazole (CDI), or oxalyl chloride.
  • Scheme IV Compounds of Structure 5:
  • Bis-acylation of compounds of structure 4 using base such as lithium hexamethyldisilazide (LiHMDS), and a disubstituted carbonate, such as dimethyl carbonate, produces compounds of structure 5.
  • base such as lithium hexamethyldisilazide (LiHMDS)
  • a disubstituted carbonate such as dimethyl carbonate
  • compounds of structure 5 can be used in the presence of the dicarbonate, for example, methyl chloroformate.
  • Scheme V describes an alternative method to prepare compounds of structure 5.
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among CpC 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C 1 -C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 halo
  • R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, CpC 8 heteroalkyl, heteroaryl and aryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, heteroaryl and aryl are optionally substituted.
  • substituents can be selected from among a subset of the listed alternatives.
  • the unsaturated ester of structure 7 was prepared by Knoevanagel condensation (see e.g., EP 0373423; and U.S. 5,032,602). For example, 229 ml (2 mol) of dimethyl malonate, 223 ml (2 mol) of 4-fluorobenzaldehyde, 40 ml of piped dine and 103 ml of glacial acetic acid are heated under reflux overnight in 1.5 1 of cyclohexane in a water separator. After cooling to room temperature, the mixture is taken up in ethyl acetate, and the solution is washed with water, dried using sodium sulphate and distilled.
  • Knoevanagel condensation see e.g., EP 0373423; and U.S. 5,032,602
  • 229 ml (2 mol) of dimethyl malonate 223 ml (2 mol) of 4-fluorobenzaldehyde, 40 ml of piped dine and 103 ml of glacial
  • Amide substrate of structure 8 was prepared by reacting acyl halides, such as methyl malonyl chloride, with an amine, such as methyl amine, under anhydrous conditions.
  • Scheme VI describes the hydrolysis of compounds of structure 5 to produce compounds of structure 10.
  • a compound of structure 9 can be an intermediate in the conversion of 5 to 10.
  • R 1 is selected from among hydrogen, OR', NR 'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, C]-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, Ci-C 8 haloalkyl, Cj-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C,-C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted; and R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C
  • the hydrolysis of esters can be achieved chemically or biochemically. Chemical hydrolysis can be selective or non-selective. Biochemical catalysis includes processes conducted by microorganisms and enzymes. Enzymatic hydrolysis, including hydrolysis by carboxy esterases and alkaline proteases, selectively cleaves ester bonds. For example, treatment of compounds of structure 5 with an appropriate enzyme solution, such as a hydrolase solution, produces compounds of structure 10. Optimization of reaction conditions include enzyme selection, medium selection, use of organic co-solvents, buffer selection, temperature, enzyme concentration, substrate concentration, and pH.
  • the hydrolase can be an esterase, a lipase, or a protease.
  • the hydrolase can be of mammalian, including human, origin, or can be of non-mammalian origin, including but not limited to, plant, bacterial, viral, yeast and fungal origin. Treatment of a compound of structure 5 with such a hydrolase enzyme affords a compound of structure 10.
  • High throughput methods can be employed to screen multiple enzymes for their ability to produce a compound of structure 10 from a compound of structure 5. Such methods are typically performed in, for example, 96-well microtiter plates, such that multiple hydrolases (e.g., a hydrolase library) can be simultaneously screened for hydrolysis of a chosen substrate and stereoselectivity.
  • hydrolases e.g., a hydrolase library
  • One of skill in the art can determine suitable conditions for the enzymatic desymmetrization of a compound of structure 5 to a compound of structure 10, taking into account the type of hydrolase being employed in the reaction.
  • the hydrolase can be a wild-type protein or a variant thereof.
  • the hydrolase variant can contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more mutations, such as amino acid substitutions, deletions, additions, or combinations thereof.
  • hydrolase variants can be generated by directed evolution such that the substrate specificity and stereoselectivity of the variants are optimized for the purposes herein (see e.g. Bornscheuer et ⁇ i, (1999) Curr. Opin. Biotech. 7:2169-2173, Reetz et ⁇ l, (2004) PNAS 101 :5716-5722).
  • the reactions can be performed in volumes of about 100 ⁇ l of a suitable buffer, such as 0.1 M potassium phosphate buffer, containing 1 mg/ml of substrate, 10 mg/ml of enzyme and 10% organic solvent.
  • a suitable buffer such as 0.1 M potassium phosphate buffer
  • the amount of hydrolase used in the reactions is determined by the activity of the enzyme.
  • An IU International Unit designates that amount of an enzyme preparation that catalyzes the formation of one micromole of product per minute. Such determinations can be made using methods well known in the art. Typically, 10 to 10,000 IU of hydrolase is added to the reaction for every gram of substrate.
  • a suitable pH for the reaction can be determined by one of skill in the art, taking into account the activity and stability of the hydrolase. A pH range from 3 to 11 is contemplated for the methods herein.
  • the pH of the reaction is generally maintained at between 7.0 and 7.4, but can be modified to, for example, pH 8.
  • a neutral environment is maintained, such that the pH of the reaction is at or about 6.7, 6.8, 6.9, 7.0, 7.1 , 7.2, 7.3, 7.4, 7.5, 7.6 or 7.7.
  • Suitable aqueous phases include, but are not limited to, buffers such as glutamic acid- glutamate, phosphoric acid-phosphate, acetic acid-acetate and citric acid-citrate buffers.
  • Water immiscible solvents such as methyl tert-butyl ether (MTBE), ethyl acetate, dichloromethane, toluene or diisopropyl ether (DIPE) can be used for lipase screening, resulting in a biphasic system.
  • Proteases and esterases can be screened in water miscible solvents, such as acetonitrile, methanol, acetone or ethanol.
  • the temperature during the reaction can be between 0 0 C to 60 0 C. In some embodiments, an ambient temperature is maintained throughout the reaction. In other embodiments, a temperature of 30 0 C or 37 0 C is maintained.
  • HPLC high performance liquid chromatography
  • CE capillary electrophoresis
  • GC gas chromatography
  • TLC thin layer chromatography
  • LC-MC liquid chromatography coupled with mass spectrometry
  • Scheme VII describes the synthesis of compounds of structures 12 and 13 from compounds of structure 10.
  • Compounds of structure 10 can be used to prepare paroxetine and related compounds of structure 13.
  • R 1 is selected from among hydrogen, OR', NR'R", Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C 1 -C 8 haloalkyl, Cj-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • R 2 is selected from among Ci-C 8 alkyl, C 2 -C 8 alkenyl, C 2 -C 8 alkynyl, C r C 8 haloalkyl, Ci-C 8 heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted;
  • X is selected from among hydrogen, F, Cl, Br, I, CN, NO 2 , OR', SR', SOR', SO 2 R', CO 2 R', NR'R", C r C 4 alkyl, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, Ci-C 4 haloalkyl, C 2 - C 4 haloalkenyl, C 2 -C 4 haloalkynyl, Ci-C 4 heteroalkyl, C 2 -C 4 heteroalkenyl and C 2 -C 4 heteroalkynyl, wherein the alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl are optionally substituted; and R' and R" each independently is selected from among hydrogen, Ci-C 8 alkyl, C 2 -C 8 alkenyl, C
  • Compounds of structure 11 are prepared by reducing compounds of structure 10 to the corresponding alcohol (see, e.g., U.S. 4,902,801). ).
  • trans-3- ethoxycarbonyl-4-(4'-fluorophenyl)-N-methylpiperidin-2,6-dione (18.0 g) in tetrahydrofuran (80 ml) was added slowly to a solution of lithium aluminium hydride (6.0 g) in tetrahydrofuran (40 ml) under nitrogen, keeping the temperature below 40°.
  • the reaction mixture was stirred at room temperature overnight, then warmed to 50° for about 7 hours and finally stirred overnight at room temperature.
  • Deprotection of the nitrogen moiety of a compound of structure 12 can produce compounds of structure 13, including paroxetine.
  • Reaction Conditions The reactions described in any of Schemes IV-VII above can be performed at various temperatures and over various temperature ranges. The temperature depends upon various parameters such as reactant and solvent selection, and can be empirically determined. For example, the reaction can be performed at room temperature (e.g., 21 0 C). The reaction can be performed below room temperature, for example, at temperatures between 5 to 0 0 C, between 0 to -10 0 C, between 0 to -20 0 C, or between 0 to -35 0 C (or lower).
  • the reaction can be performed at temperatures greater than room temperature, for example, at temperatures between 40-250 0 C (or higher).
  • the reaction can be performed between 40 to 80 0 C, between 80 to 120 0 C, between 120 to 180 0 C, or between 150 to 250 0 C.
  • the reaction can be conducted at temperatures between 65-70 0 C.
  • the reactions can be performed in various solvents and solvent combinations, including organic solutions, aqueous solutions and combinations thereof.
  • a suitable solvent or solvent mixture can be determined empirically by one of skill in the art, taking into account the properties of the reactants.
  • the solvent can be an organic protic or aprotic solvent including, but not limiting to, an alcohol, such as methanol, ethanol, isopropanol, butanol, pentanol or hexanol, acetone, ethyl acetate, dithianes, THF, dioxane, acetonitrile, nitromethane, dichloromethane, dichloroethane, diethyl ether, dimethylformamide (DMF), dimethylsulfoxide (DMSO), dimethyl-acetamide, or any combination thereof.
  • the reaction can be conducted in solvent and solvent combinations such as THF, MeOH, THF/TEA, THF/NaOtBu and MeOH/TEA.
  • the reactions can be performed with reactants at dilute or concentrated levels.
  • the reactants can be at concentrations between about 0.1 - 5 nmol/L, 0.01-0.5 mmol/L, 1-10 mmol/L, or 0.1-5 mol/L.
  • the reaction can be performed over any time period.
  • the reaction can be conducted for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58 or 59 minutes.
  • the reaction can be conducted for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours.
  • the reaction can be conducted over a period of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 days.
  • concentration and time for the reaction depend on various parameters such as temperature, solvent, and reactants, and can be empirically determined.
  • the reactions can be performed under various pressures. The pressure depends upon various parameters including reactant or catalyst selection and can be empirically determined.
  • the reaction can be conducted at normal atmospheric pressure (e.g., 1 arm).
  • the reaction can be carried out under pressures higher than atmospheric pressure, for example, at pressures of 2 - 10 arms, 10 - 50 arms, 50 - 100 atms, or 100 - 200 atms.
  • the reaction can be conducted at pressures lower than atmospheric pressure, for example, at pressures of 0.9 - 0.5 atms or 0.5 - 0.1 atms.
  • the reactions can be homogeneous or heterogeneous.
  • the reactions can be a bi- phasic mixture or an emulsion, or the reactants can be immobilized to a solid support.
  • the solid support can be selected from among metal, ceramic or plastic plates, beads, microbeads, membranes, filaments, microtitre trays, and the wall of a reaction chamber.
  • suitable solid supports include semi-permeable membranes, glass capillaries, alumina, alumina supported polymers, silica, chemically bonded hydrocarbons on silica, polyolefins, agarose, polysaccharides such as dextran, or glycoproteins such as fibronectin.
  • alumina alumina supported polymers
  • silica chemically bonded hydrocarbons on silica
  • polyolefins such as polyolefins
  • agarose polysaccharides
  • polysaccharides such as dextran
  • glycoproteins such as fibronectin
  • the compounds provided herein can possess sufficiently acidic or sufficiently basic functional group(s) to form pharmaceutically acceptable salts.
  • Pharmaceutically acceptable salts are obtained using standard procedures well known in the art. For example, if the compound includes an acidic functionality, a pharmaceutically acceptable salt is prepared by any suitable method, such as by treatment of the acid with an inorganic or organic base. If the compound has a basic functionality, a pharmaceutically acceptable salt is prepared by any suitable method, such as by treatment of the base with an inorganic or organic acid.
  • a single compound provided herein can include more than one acidic or basic moiety and can include mono, di or tri-salts.
  • Compounds of Formula I, I A , II, VI, VII, VIII, IX and X can possess a sufficiently acidic functional group to form a pharmaceutically acceptable salt.
  • Exemplary salts can be formed by treatment or these compounds with metals and alkali metals (for example, sodium, potassium or lithium), alkaline-earth metal hydroxides or alkoxides (such as ethoxide or methoxide) and transition metals (such as magnesium or aluminum).
  • compounds with a suitably acidic proton can be treated with organic bases (such as ammonia or secondary or tertiary amines, including diethylamine, triethylamine, piperidine, piperazine, morpholine, choline, or meglumine), with basic amino acids, or with amino alcohols (such as 3-aminobutanol and 2-aminoethanol).
  • organic bases such as ammonia or secondary or tertiary amines, including diethylamine, triethylamine, piperidine, piperazine, morpholine, choline, or meglumine
  • basic amino acids such as 3-aminobutanol and 2-aminoethanol
  • Paroxetine and compounds of Formula I, I A , II, VI, VII, VIII, IX and X can possess a sufficiently basic functional group to form a pharmaceutically acceptable salt.
  • Exemplary salts can be formed by treatment of these compounds with organic acids, such as acetic acid, adamantanecarboxylic acid, adipic acid, ascorbic acid, aspartic acid, azelaic acid, benzoic acid, 2-(4-hydroxybenzoyl) benzoic acid, carboxylic, cinnamic acid, citric acid, cyclohexanecarboxylic acid, decanoic acid, dodecanoic acid, 1 ,2-ethanedisulphonic acid, ethanesulphonic acid, ethylenediamine- tetraacetic acid (EDTA), fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hippuric acid, lactic acid, lactobionic acid, maleic acid, hydroxy
  • Inorganic acid addition salts also can be formed, including, but not limited to, bicarbonates, carbonates, chlorides, bromides, iodides, nitrates, perchlorates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, sulfates, pyrosulfates, bisulfates, sulfites, and bisulfites.
  • bicarbonates carbonates, chlorides, bromides, iodides, nitrates, perchlorates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, sulfates, pyrosulfates, bisulfates, sulfites, and bisulfites.
  • the methods provided herein for the enzymatic production of paroxetine and/or the production of racemic and/or enantiomerically enriched intermediates used in the synthesis of paroxetine can be performed at any scale.
  • an appropriate substrate as described herein is dissolved or suspended in an appropriate solvent, such as water or co-solvent system, in a 500 mL to 2 L conical flask or 1-5 gallon reactor followed by addition of 10 to 10,000 IU of a hydrolase for every gram of substrate.
  • the reaction mixture is stirred using an incubator shaker or a blade or propeller mixer for the time required for conversion of the substrate to the product (1-96 hrs), and the product is recovered and the structure confirmed by known spectral data.
  • the enzyme can be recovered and reused by separation of the reaction product, such as by ultrafiltration, which includes passing the solvent containing the product through an ultrafiltration membrane that retains compounds with molecular weights higher than the reaction product, such as the hydrolase, while allowing the substrate to pass through for collection, such as by extraction, evaporation, concentration, isolation or other technique known in the art.
  • ultrafiltration includes passing the solvent containing the product through an ultrafiltration membrane that retains compounds with molecular weights higher than the reaction product, such as the hydrolase, while allowing the substrate to pass through for collection, such as by extraction, evaporation, concentration, isolation or other technique known in the art.
  • Any ultrafiltration cartridge known in the art having the appropriate molecular weight cutoff for separating enzyme from the product in the product stream can be used.
  • a batch, semi-continuous or continuous process can be used.
  • the enzyme can be free in solution or can be immobilized on a solid support.
  • it efficiently can be recovered and recycled, such as by using an ultrafiltration membrane selected to retain the enzyme due its high molecular weight relative to the molecular weights of the substrate and product.
  • the enzyme can be retained in a reaction chamber while low molecular weight product and unreacted substrate passes through the ultrafiltration membrane. The reaction product is removed from the effluent stream, additional substrate is added to the product-free effluent stream and the effluent stream is returned to the reaction chamber.
  • the enzyme also can be attached to a solid support.
  • the solid support can be selected from among glass, plastic, polystyrene, polycarbonate, polypropylene, polyethylene, nylon, a sol-gel polymer, chitin, sand, pumice, agarose, dendrimers, buckyballs, polyacrylamide, silicon, rubber, latex, semi- or partially permeable membranes, such as of regenerated cellulose or cellophane, cellulose, nitrocellulose, cellulose acetate, agarose polyacrylamide copolymer, starch, nylon polyesters, dextran, cross-linked dextran, dextran acrylamide copolymer, cyclodextran, cross- linked hydroxyethylmethacrylate substituted cross-linked polystyrenes, polyvinylalcohol, celite and silica.
  • the method can be performed in a membrane reactor, which includes a reaction vessel in which the enzyme catalyst is attached to a solid support, while low-molecular substances within the reactor are able to leave the reactor.
  • the membrane is integrated directly into the reaction chamber.
  • the membrane is incorporated outside the reactor chamber in a separate filtration module, with the reaction solution flowing continuously or intermittently through the filtration module, and with the retentate being recirculated into the reactor.
  • a continuous membrane reactor the product is continuously removed and a continuous amount of substrate is provided.
  • the method can use micro-porous or semi-permeable membranes, including those enclosed in a cartridge, that are placed in a circuit in which the solvent including the substrate recycles after removal of the resulting product.
  • the enzyme is immobilized on a solid support, such as a micro-porous or semi-permeable membrane, and the enzyme contacts the solvent containing the substrate at the membrane-enzyme interface.
  • the immobilized enzyme converts the substrate to product, and the solution from which the product is recovered can be replenished with substrate and returned to a substrate vessel or to the enzyme membrane reactor.
  • the circuit optionally can include a reactor circulating pump that pumps the solution leaving the enzyme reactor vessel to a product recovery vessel.
  • micro-porous or semi-permeable membrane can be used as a solid support.
  • Perforated membranes are considered to be "micro-porous" if they have pore sizes down to a few nm, particularly, 10-50 nm, and in particular 1-10 nm.
  • an ultrafiltration membrane with pore sizes of ⁇ 1 nm has a molecular weight cutoff of 1000 daltons. The upper limit is not critical so long as it provides an appropriate molecular weight cutoff to allow separation of the product from the reactants, such as the enzyme.
  • the method can be performed using a semi-permeable membrane with pore sizes of 1 nm, 5 nm, 10 nm, 25 nm, 50 nm, 100 nm, 1 ⁇ m, 5 ⁇ m, 10 ⁇ m and larger.
  • Any micro- porous or semi-permeable membrane including hydrophobic and hydrophilic membrane, can be utilized, including a hollow fiber membrane in any configuration, such as a bundle. Hollow fibers provide large membrane surfaces and small liquid layer membrane surfaces and small liquid layer thicknesses can be realized. This arrangement allows the velocity of the flow of the substrate-bearing solvent to be controlled to a rate appropriate for the velocity of enzymatic product formation. Hollow fibers made of any suitable material can be used.
  • Exemplary hydrophobic materials include hydrocarbon polymers, such as polypropylene and polyethylene, and fluoridated derivatives thereof, whose relatively high porosity and small wall thickness with relatively small pore sizes make them particularly suitable for use in the membrane reactor.
  • the size and the number of the pores that pass through the solid support material can be selected to maximize the rate of transfer (flux) of the substrate across the membrane and in contact with the enzyme to yield product. It will be appreciated that the pore size can be varied depending on the properties of the immobilized enzyme, reactants employed, products produced, and reaction factors such as pH, concentration, pressure and temperature; and further, that the optimum pore size and flow rates can be determined empirically by those skilled in the art. The pH and temperature of the reaction and product phases are maintained at a value that keeps the reactants in a form that maximizes enzymatic product formation. A useful discussion of pore size selection is found in U.S. Pat. No. 4,174,374.
  • membrane reactors examples include U.S. Pat. Nos. 7,198,941, 6,942,799, 6,107,055, 5,077,217, 5,057,421, 4,963,494, 4,861,483, 4,795,704, 4,442,206, 4,187,086.
  • Enzyme membrane reactors of various configuration and sizes, from bench scale to full-scale industrial units, are well known to the skilled artisan (for a discussion of which, see, e.g., Biocatalysis: Fundamentals and Applications, Andreas S. Bommarius and Bettina R.
  • the bis-isopropyl ester (Compound 110, Structure 6, Scheme IV) was synthesized from Compound 104 (Structure 5, Scheme IV).
  • Catalytic KCN was used as a transesterification catalyst with heating.
  • An enzyme solution was prepared as follows: the protease subtilisin Carlsberg obtained from Sigma-Aldrich Co. (Saint Louis, MO, Cat. No. P-5380) (250 mL liquid formulation containing approximately 25 g of enzyme) was dialyzed against potassium phosphate buffer (10 mM, pH 7.5) containing 0.025 mM calcium acetate. The resulting enzyme solution (350 mL) was then used directly in the reaction.
  • Relative conversion % HPLC peak area of monoester product / Sum of HPLC peak areas of monoester product and bis-ester starting material
  • the identified enzymes fall into several categories of hydrolases including lipases, esterases and proteases.
  • the identified enzymes that gave the most conversions under the screening conditions were carboxyl-esterases (EC 3.1.1.1), including pig and rabbit liver esterases, and alkaline proteases (EC 3.4.21.62), such as subtilisin Carlsberg.
  • the sequences identifiers (SEQ ID NOS) corresponding to the amino acid sequences of these and other exemplary hydrolases are provided in Table 3.
  • exemplary carboxylesterases including pig and rabbit liver esterase, are set forth in SEQ ID NOS: 1-3 and 28-113; exemplary subtilisins, including subtilisin Carlsberg, are set forth in SEQ ID NOS:4-8 and 15-27; exemplary oryzins (i.e. EC 3.4.21.63) are set forth in SEQ ID NOS:9-14; exemplary endopeptidases are set forth in SEQ ID NOS:114-172; and exemplary lipases are set forth in SEQ ID NOS:173-187.
  • Co-solvent screening for enzymatic hydrolysis of Compound 104 to Compound 118 Medium optimization is an important part of enzymatic reaction engineering for improving both enzyme reactivity and selectivity.
  • Pig liver esterase (PLE) proved to be less tolerant to the organic co-solvents, resulting in either lower reactivity or deactivation.
  • no reversal of its enantioselectivity was observed.
  • subtilisin Carlsberg was very tolerant to a number of solvents and significant improvement of reactivity was achieved. The following summarizes the results from the solvent screening.
  • the substrate Compound 104, 1 mg/mL
  • the enzyme subtilisin Carlsberg P-5380 (Sigma), 5 mg/mL
  • pH 6.5 the substrate
  • the reactions were run in a 96-well plate at 40 °C with vigorous mixing (900 rpm). After 18-24 h, the reactions were stopped by the addition of three volumes of acetonitrile, and analyzed by HPLC. Relative percent conversions were calculated as described in Example 19. In general, the enzyme either lost activity partially or completely at 50% organic co-solvents (with the exception of MTBE). Another observation was that substrate degradation was less severe at higher organic contents, presumably due to better protection of the substrate by organic solvents.
  • Method A DMSO was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% DMSO, the relative percent conversion of compound 104 to 118 was 10 percent. At 30% DMSO, the relative percent conversion of compound 104 to 118 was 80 percent. At 50% DMSO, the relative percent conversion of compound 104 to 118 was 40 percent.
  • Method B Acetone was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% acetone, the relative percent conversion of compound 104 to 118 was 30 percent. At 30% acetone, the relative percent conversion of compound 104 to 118 was 50 percent. At 50% acetone, the relative percent conversion of compound 104 to 118 was 20 percent.
  • Method C t-Butanol was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% t-butanol, the relative percent conversion of compound 104 to 118 was 90 percent. At 30% t-butanol, the relative percent conversion of compound 104 to 118 was 67 percent. At 50% t-butanol, the enzyme lost activity partially or completely.
  • THF was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% THF, the relative percent conversion of compound 104 to 118 was 30 percent. At 30% THF, the relative percent conversion of compound 104 to 118 was 81 percent. At 50% THF, the enzyme lost activity partially or completely.
  • Method E Dioxane was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% dioxane, the relative percent conversion of compound 104 to 118 was 70 percent. At 30% dioxane, the relative percent conversion of compound 104 to 118 was 50 percent. At 50% dioxane, the enzyme lost activity partially or completely.
  • Method F t-Pentanol was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% t-pentanol, the relative percent conversion of compound 104 to 118 was 93 percent. At 30% t-pentanol, the relative percent conversion of compound 104 to 118 was 17 percent. At 50% t-pentanol, the enzyme lost activity partially or completely.
  • Method G MTBE was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 10% MTBE, the relative percent conversion of compound 104 to 118 was 14 percent. At 30% MTBE, the relative percent conversion of compound 104 to 118 was 28 percent. At 50% MTBE, the relative percent conversion of compound 104 to 118 was 64 percent.
  • Organic co-solvent content was further varied based on the results from the initial solvent screen. Solvents that gave higher conversion from the initial screen were selected and the organic co-solvent contents were evaluated over smaller ranges. The reactions were run at a substrate concentration of lmg/mL of 104 and at a reduced enzyme loading (1 mg/mL enzyme) with pH 6.0 or 6.5 for 16 hours at 40 °C.
  • Method H (pH 6.0): DMSO was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 20% DMSO, the relative percent conversion of compound 104 to 118 was 24 percent. At 25% DMSO, the relative percent conversion of compound 104 to 118 was 20 percent. At 30% DMSO, the relative percent conversion of compound 104 to 118 was 20 percent. At 35% DMSO, the relative percent conversion of compound 104 to 118 was 15 percent. At 40% DMSO, the relative percent conversion of compound 104 to 118 was 14 percent.
  • Method I (pH 6.5): DMSO was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 20% DMSO, the relative percent conversion of compound 104 to 118 was 30 percent. At 25% DMSO, the relative percent conversion of compound 104 to 118 was 32 percent. At 30% DMSO, the relative percent conversion of compound 104 to 118 was 30 percent. At 35% DMSO, the relative percent conversion of compound 104 to 118 was 30 percent. At 40% DMSO, the relative percent conversion of compound 104 to 118 was 20 percent.
  • Method J (pH 6.0): Acetone was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104.
  • the relative percent conversion of compound 104 to 118 was 8 percent. At 25% acetone, the relative percent conversion of compound 104 to 118 was 9 percent. At 30% acetone, the relative percent conversion of compound 104 to 118 was 5 percent. At 35% acetone, the relative percent conversion of compound 104 to 118 was 4 percent. At 40% acetone, the relative percent conversion of compound 104 to 118 was 2 percent. Method K (pH 6.5); Acetone was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 20% acetone, the relative percent conversion of compound 104 to 118 was 4 percent. At 25% acetone, the relative percent conversion of compound 104 to 118 was 18 percent.
  • Method L THF was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 20% THF, the relative percent conversion of compound 104 to 118 was 8 percent. At 25% THF, the relative percent conversion of compound 104 to 118 was 7 percent. At 30% THF, the relative percent conversion of compound 104 to 118 was 2 percent. At 35% THF, the relative percent conversion of compound 104 to 118 was 1 percent. At 40% THF, the relative percent conversion of compound 104 to 118 was 1 percent.
  • Method M (pH 6.5): THF was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104.
  • the relative percent conversion of compound 104 to 118 was 15 percent.
  • the relative percent conversion of compound 104 to 118 was 16 percent.
  • the relative percent conversion of compound 104 to 118 was 8 percent.
  • the relative percent conversion of compound 104 to 118 was 3 percent.
  • the relative percent conversion of compound 104 to 118 was 2 percent.
  • Method N (pH 6.0): t-Butanol was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 5% t-butanol, the relative percent conversion of compound 104 to 118 was 2 percent. At 10% t-butanol, the relative percent conversion of compound 104 to 118 was 19 percent. At 15% t- butanol, the relative percent conversion of compound 104 to 118 was 9 percent. At 20% t-butanol, the relative percent conversion of compound 104 to 118 was 12 percent. At 25% t-butanol, the relative percent conversion of compound 104 to 118 was 7 percent. Method O (pH 6.5): t-Butanol was used as a co-solvent for subtilisin
  • Method P (pH 6.0): t-Pentanol was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 5% t-pentanol, the relative percent conversion of compound 104 to 118 was 17 percent. At 10% t-pentanol, the relative percent conversion of compound 104 to 118 was 21 percent. At 15% t- pentanol, the relative percent conversion of compound 104 to 118 was 12 percent. At 20% t-pentanol, the relative percent conversion of compound 104 to 118 was 4 percent. At 25% t-pentanol, the relative percent conversion of compound 104 to 118 was 3 percent.
  • Method Q (pH 6.5): t-Pentanol was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 5% t-pentanol, the relative percent conversion of compound 104 to 118 was 36 percent. At 10% t-pentanol, the relative percent conversion of compound 104 to 118 was 40 percent. At 15% t- pentanol, the relative percent conversion of compound 104 to 118 was 26 percent. At 20% t-pentanol, the relative percent conversion of compound 104 to 118 was 6 percent. At 25% t-pentanol, the relative percent conversion of compound 104 to 118 was 4 percent.
  • Method R (pH 6.0): Dioxane was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 5% dioxane, the relative percent conversion of compound 104 to 118 was 2 percent. At 10% dioxane, the relative percent conversion of compound 104 to 118 was 11 percent. At 15% dioxane, the relative percent conversion of compound 104 to 118 was 11 percent. At 20% dioxane, the relative percent conversion of compound 104 to 118 was 10 percent. At 25% dioxane, the relative percent conversion of compound 104 to 118 was 7 percent.
  • Method S (pH 6.5): Dioxane was used as a co-solvent for subtilisin Carlsberg catalyzed hydrolysis of compound 104. At 5% dioxane, the relative percent conversion of compound 104 to 118 was 16 percent. At 10% dioxane, the relative percent conversion of compound 104 to 118 was 20 percent. At 15% dioxane, the relative percent conversion of compound 104 to 118 was 19 percent. At 20% dioxane, the relative percent conversion of compound 104 to 118 was 15 percent. At 25% dioxane, the relative percent conversion of compound 104 to 118 was 12 percent.
  • Method B At 30% DMSO, substrate concentration of 5 mg/mL 104, and enzyme concentration of 25 mg/mL, the product yield of 118 was 65 percent. There was 3 percent substrate remaining.
  • Method C At 10% t-pentanol, substrate concentration of 5 mg/mL 104, and enzyme concentration of 25 mg/mL, the product yield of 118 was 26 percent. There was 75 percent substrate remaining.
  • Method D At 20% DMSO, substrate concentration of 10 mg/mL 104, and enzyme concentration of 50 mg/mL, the product yield of 118 was 32 percent. There was 25 percent substrate remaining.
  • Method E At 30% DMSO, substrate concentration of 10 mg/mL 104, and enzyme concentration of 50 mg/mL, the product yield of 118 was 42 percent. There was 18 percent substrate remaining.
  • Method F At 10% t-pentanol, substrate concentration of 10 mg/mL 104, and enzyme concentration of 50 mg/mL, the product yield of 118 was 13 percent. There was 55 percent substrate remaining.
  • Subtilisin Carlsberg also presumably catalyzed transesterification of the substrate with methanol, ethanol and iso-propanol co-solvent (products have not been characterized).
  • phosphate buffer 50 mM, pH 6.5
  • 10% t-pentanol was chosen as the medium for the hydrolysis.
  • 107 showed a significant amount of amide ring opening that lowered the overall yield.
  • Both 108 and 109 were substantially more soluble in buffer and were therefore selected for further investigation of the reaction at higher substrate concentrations.
  • the hydrolysis reactions of 108 and 109 were carried out at 0.5 mL volume in 2 mL glass HPLC vials at 35 °C.
  • the substrate concentration was 10 mg/mL in potassium phosphate buffer (50 mM, pH 6.5) with 10% t-pentanol.
  • the reaction was initiated by the addition of P-5380 (Sigma) to a concentration of 10 mg/mL.
  • An aliquot (10 ⁇ L) of the reaction mixture was removed from the reaction every 3 hours and quenched with 290 ⁇ L of acetonitrile. Brief centrifugation was used to remove the precipitates and the supernatant was analyzed using HPLC.
  • a negative control with no enzyme added was used to evaluate non-catalyzed hydrolysis of the two substrates. In comparison with hydroxyethyl 108, hydrolysis of hydroxypropyl 109 proceeded more slowly. However, 109 was more stable to non-catalyzed hydrolysis than 108.
  • Method A Subtilisin Carlsberg catalyzed hydrolysis of the hydroxyethyl substrate 108 in potassium phosphate buffer at pH 6.5. The non-catalyzed hydrolysis of both the ester and imide bonds was estimated at approximately 10%. After 3 hours the approximate yield of product was 22 percent and the remaining substrate was approximately 65 percent. After 6 hours, the approximate yield of product was 37 percent and the remaining substrate was approximately 45 percent. After 9 hours, the approximate yield of product was 59 percent and the remaining substrate was approximately 27 percent. At 12 hours, the approximate yield of product was 63 percent and the remaining substrate was approximately 23 percent. The reactions were terminated after 12 hours and the product was extracted with MTBE. The enantiomeric excess of the product was analyzed using chiral HPLC method. The ee for the product was approximately 70 percent.
  • Method B Subtilisin Carlsberg catalyzed hydrolysis of the hydroxypropyl substrate 109 in potassium phosphate buffer at pH 6.5. Substrate 109 was more stable to non-catalyzed hydrolysis than 108. The background hydrolysis of 109 was negligible. After 3 hours the approximate yield of product was 19 percent and the remaining substrate was approximately 67 percent. After 6 hours, the approximate yield of product was 26 percent and the remaining substrate was approximately 59 percent. After 9 hours, the approximate yield of product was 44 percent and the remaining substrate was approximately 45 percent. At 12 hours, the approximate yield of product was 46 percent and the remaining substrate was approximately 40 percent. The reactions were terminated after 12 hours and the product was extracted with MTBE.
  • the enantiomeric excess of the product was analyzed using chiral HPLC method.
  • the ee for the product was approximately 89 percent.
  • the higher enantiomeric excess of the product is likely due to the lower background ester hydrolysis of 109 than that of hydroxyethyl 108.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des composés et des procédés qui produisent des intermédiaires et des précurseurs de la paroxétine et composés associés. L'invention concerne également des procédés de préparation de la paroxétine.
PCT/US2008/007870 2007-06-27 2008-06-24 Composés et procédé de préparation d'intermédiaires chiraux pour la synthèse de la proxétine WO2009005647A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US93746607P 2007-06-27 2007-06-27
US60/937,466 2007-06-27

Publications (2)

Publication Number Publication Date
WO2009005647A2 true WO2009005647A2 (fr) 2009-01-08
WO2009005647A3 WO2009005647A3 (fr) 2009-04-09

Family

ID=40076855

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/007870 WO2009005647A2 (fr) 2007-06-27 2008-06-24 Composés et procédé de préparation d'intermédiaires chiraux pour la synthèse de la proxétine

Country Status (1)

Country Link
WO (1) WO2009005647A2 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013136057A3 (fr) * 2012-03-13 2013-11-14 Glaxosmithkline Australia Pty Limited Molécule d'acide nucléique
CN104892491A (zh) * 2015-05-06 2015-09-09 浙江海森药业有限公司 一种合成帕罗西汀手性中间体的方法
US9200261B2 (en) 2010-07-22 2015-12-01 Glaxosmithkline Australia Pty Limited Plant cytochrome P450
CN105400758A (zh) * 2015-11-26 2016-03-16 青岛康原药业有限公司 一种氯化钠-氨水、丙酮协同纯化胰激肽酶原的方法及提高胰激肽酶原稳定性的药物组合物
CN105418502A (zh) * 2014-09-22 2016-03-23 浙江九洲药业股份有限公司 一种用于合成帕罗西汀的中间体及其制备方法和用途
US9458481B2 (en) 2010-06-22 2016-10-04 Sun Pharmaceutical Industries (Australia) Pty Ltd Methyltransferase nucleic acids and polypeptides
WO2021067627A1 (fr) * 2019-10-01 2021-04-08 Mars, Incorporated Traitement enzymatique d'anthocyanines
CN114214375A (zh) * 2021-12-27 2022-03-22 江苏威凯尔医药科技有限公司 (r)-3-(4-卤代-1h-吡唑-1-基)-3-环戊基丙酸酯的制备方法
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes
US11986006B2 (en) 2018-08-31 2024-05-21 Mars, Incorporated Anthocyanin-based colorant compositions and methods of use thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993022284A1 (fr) * 1992-05-06 1993-11-11 Smithkline Beecham Plc Procede d'hydrolyse stereospecifique de derives de piperidinedione

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993022284A1 (fr) * 1992-05-06 1993-11-11 Smithkline Beecham Plc Procede d'hydrolyse stereospecifique de derives de piperidinedione

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
D. A. GREENHALGH ET AL.: "A Novel Chiral Base Mediated Glutarimide Desymmetrisation: Application to the Asymmetric Synthesis of (-)-Paroxetine" SYNLETT, no. 12, 2 December 2002 (2002-12-02) , pages 2074-2076, XP002513234 GEORG THIEME VERLAG, STUTTGART; DE ISSN: 0936-5214 cited in the application *

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9458481B2 (en) 2010-06-22 2016-10-04 Sun Pharmaceutical Industries (Australia) Pty Ltd Methyltransferase nucleic acids and polypeptides
US11312973B2 (en) 2010-07-22 2022-04-26 Sun Pharmaceutical Industries (Australia) Pty Ltd Plant cytochrome P450
US9200261B2 (en) 2010-07-22 2015-12-01 Glaxosmithkline Australia Pty Limited Plant cytochrome P450
US10844391B2 (en) 2010-07-22 2020-11-24 Sun Pharmaceutical Industries (Australia) Pty Ltd Plant cytochrome P450
US10385354B2 (en) 2010-07-22 2019-08-20 Sun Pharmaceutical Industries (Australia) Pty Ltd Papaver somniferum cytochrome P450
US9725732B2 (en) 2010-07-22 2017-08-08 Sun Pharmaceutical Industries (Australia) Pty Ltd Plant cytochrome P450
US9447444B2 (en) 2012-03-13 2016-09-20 Sun Pharmaceutical Industries (Australia) Pty Ltd Biosynthesis of opiate alkaloids
WO2013136057A3 (fr) * 2012-03-13 2013-11-14 Glaxosmithkline Australia Pty Limited Molécule d'acide nucléique
US9862979B2 (en) 2012-03-13 2018-01-09 Sun Pharmaceutical Industries (Australia) Pty Ltd Biosynthesis of opiate alkaloids
CN105418502B (zh) * 2014-09-22 2018-12-14 浙江九洲药业股份有限公司 一种用于合成帕罗西汀的中间体及其制备方法和用途
CN105418502A (zh) * 2014-09-22 2016-03-23 浙江九洲药业股份有限公司 一种用于合成帕罗西汀的中间体及其制备方法和用途
CN104892491A (zh) * 2015-05-06 2015-09-09 浙江海森药业有限公司 一种合成帕罗西汀手性中间体的方法
CN104892491B (zh) * 2015-05-06 2017-05-17 浙江海森药业有限公司 一种合成帕罗西汀手性中间体的方法
CN105400758A (zh) * 2015-11-26 2016-03-16 青岛康原药业有限公司 一种氯化钠-氨水、丙酮协同纯化胰激肽酶原的方法及提高胰激肽酶原稳定性的药物组合物
US11986006B2 (en) 2018-08-31 2024-05-21 Mars, Incorporated Anthocyanin-based colorant compositions and methods of use thereof
WO2021067627A1 (fr) * 2019-10-01 2021-04-08 Mars, Incorporated Traitement enzymatique d'anthocyanines
CN115135169A (zh) * 2019-10-01 2022-09-30 马斯公司 花青素苷的酶处理
US11753664B2 (en) 2019-10-01 2023-09-12 Mars, Incorporated Enzymatic treatment of anthocyanins
CN114214375B (zh) * 2021-12-27 2024-03-19 江苏威凯尔医药科技有限公司 (r)-3-(4-卤代-1h-吡唑-1-基)-3-环戊基丙酸酯的制备方法
CN114214375A (zh) * 2021-12-27 2022-03-22 江苏威凯尔医药科技有限公司 (r)-3-(4-卤代-1h-吡唑-1-基)-3-环戊基丙酸酯的制备方法
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Also Published As

Publication number Publication date
WO2009005647A3 (fr) 2009-04-09

Similar Documents

Publication Publication Date Title
WO2009005647A2 (fr) Composés et procédé de préparation d'intermédiaires chiraux pour la synthèse de la proxétine
WO2008127646A2 (fr) Procédés à base de transaminase utilisés dans la préparation de la prégabaline
WO2009009117A2 (fr) Procédés chimio-enzymatique pour la préparation du lévétiracétam
Koul et al. Enzymatic resolution of naproxen
KR20050053618A (ko) 에스시탈로프람의 제조에 사용할 수 있는 중간체들의 분리방법
Wallace et al. Resolution of a chiral ester by lipase-catalyzed transesterification with polyethylene glycol in organic media
ES2333201T3 (es) Butinol 1 esterasa.
Miyazawa et al. Resolution of racemic carboxylic acids via the lipase‐catalyzed irreversible transesterification of vinyl esters
WO2008140127A1 (fr) Procédé destiné à produire un 2-alkyl-1,1,3-trialkoxycarbonylpropane optiquement actif
Sakai et al. Practical Resolution of 3-Phenyl-2 H-azirine-2-methanol at Very Low Temperature by Using Lipase Immobilized on Porous Ceramic and Optimized Acylating Agent
Sicart et al. Fluorogenic substrates for lipases, esterases, and acylases using a TIM‐mechanism for signal release
TW201835330A (zh) (±)-2-(二氟甲基)-1-(烷氧羰基)-環丙烷甲酸及(±)-2-(乙烯基)-1-(烷氧羰基)-環丙烷甲酸之酶催化製法
US6153414A (en) Method for racemic biochemical resolution of CIS-and trans-pyprolopiperidine
EP1487990A2 (fr) Procede de preparation d'acides beta-aminocarboxyliques optiquement actifs a partir d'acides beta-aminocarboxyliques n-acyles racemiques
US5841001A (en) Process for the preparation of optically active 2-halo-1-(substituted phenyl) ethanol
EP2796562A1 (fr) Procédé de fabrication d'un acide -substitué- -aminé optiquement actif
JPH1033191A (ja) 光学活性３−ｎ置換アミノイソ酪酸類およびその塩ならびにそれらの製造方法
Lattmann et al. Screening and application of microbial esterases for the enantioselective synthesis of chiral glycerol derivatives
WO1999004028A1 (fr) Procede de preparation d'un acide alpha-trifluoromethylacetique optiquement actif et des esters antipodes dudit acide, et methode de purification dudit acide
Chen et al. Altering lipase activity and enantioselectivity in organic media using organo‐soluble bases: Implication for rate‐limiting proton transfer in acylation step
EP1737818A2 (fr) Procedes pour la preparation d'amines stereoisomeriquement enrichies
JPH05227991A (ja) 光学活性な３−ピロリジノール誘導体の製造法
JP2006517393A5 (fr)
JP5953429B2 (ja) α−置換システインもしくはその塩又はα−置換システイン合成中間体の製造方法
JPH10273466A (ja) ２−ヒドロキシ−１−インダノンの製造方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08768749

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC, EPO FORM 1205A DATED 15.04.2010

122 Ep: pct application non-entry in european phase

Ref document number: 08768749

Country of ref document: EP

Kind code of ref document: A2