Enzymatic processes for the preparation of (±)-2-(difluoromethyl)-1-(alkoxycarbonyl)-cyclopropanecarboxylic acid and (±)-2-(vinyl)-1-(alkoxycarbonyl)-cyclopropanecarboxylic acid

Patent metadata

Field	Value
Publication number	US-2018258451-A1
Application number	US-201815886318-A
Country	US
Kind code	A1
Filing date	Feb 1, 2018
Priority date	Feb 1, 2017
Publication date	Sep 13, 2018
Grant date	—

Abstract

Disclosed are methods of synthesizing racemic 2-(difluoromethyl)-1-(alkoxycarbonyl)-cyclopropanecarboxylic acids and 2-(vinyl)-1-(alkoxycarbonyl)-cyclopropanecarboxylic acids and their salts, such as the dicyclohexylamine salt. Also disclosed are methods for preparing enantioenriched (1R,2R)-1-((tert-butoxycarbonyl)amino)-2-(difluoromethyl)cyclopropane-1-carboxylic acid and esters of the same. These compounds are useful intermediates in the synthesis of viral protease inhibitors.

First claim

1 . A method according to reaction Scheme A:

wherein R 1 is (C 1 -C 6 )alkyl;
R 2 is —CF 2 H or —CH═CH 2 ; and
the first enzyme is a mutant of BsteE esterase.

2 - 9 . (canceled)

10 . The method of claim 1 , wherein the mutant of BsteE esterase has a first mutation of T25H, a second mutation of L92H, and a third mutation selected from the group consisting of C115S, C115A, K121R, K121Y, K121H, K121D, K121L, K121T, K121A, K121N, E123T, M126F, M126V, M126I, M126Q, M126N, K164S, L166N, L166M, L166I, L166T, L166A, I170N, I170Q, I170V, I170T, I170A, D172N, M194A, M194L, I195G, I195A, I195V, and K215N.

11 - 14 . (canceled)

15 . The method of claim 1 , wherein the loading of the first enzyme is about 50 wt % to about 200 wt % as compared to the starting material in Scheme A.

16 . The method of claim 1 , wherein the first solvent comprises one or more organic solvents.

17 . (canceled)

18 . The method of claim 1 , wherein the first solvent further comprises an aqueous buffer.

19 . The method of claim 1 , wherein the first solvent further comprises a base.

20 . The method of claim 19 , wherein the base is a di(alkyl)amine, such as a di(cycloalkyl)amine or a di(aralkyl)amine.

21 - 24 . (canceled)

25 . The method of claim 1 , further comprising contacting the reaction product of reaction Scheme A with a base to obtain a salt of the compound.

26 . The method of claim 25 , wherein the base is a di(alkyl)amine, such as a di(cycloalkyl)amine or a di(aralkyl)amine.

27 . (canceled)

28 . The method of claim 1 , wherein the diastereomeric excess of the reaction product is greater than about 90%.

29 . (canceled)

30 . The method of claim 1 , further comprising the step according to reaction Scheme B:

wherein:
R 2 is —CF 2 H; and
the second enzyme is selected from the group consisting of Alcalase® 2.4 L, Esperase® 8.0 L, and Savinase® 16.0 L.

31 . (canceled)

32 . The method of claim 30 , wherein the loading of the second enzyme is about 50 wt % to about 200 wt % as compared to the starting material in Scheme B.

33 . The method of claim 30 , wherein the second solvent comprises an aqueous buffer.

34 . (canceled)

35 . The method of claim 33 , wherein the second solvent further comprises an organic solvent such as acetone.

36 - 38 . (canceled)

39 . A polypeptide, comprising an amino acid sequence having at least two amino acid substitutions relative to SEQ ID NO: 2, wherein the amino acid sequence comprises a first mutation of T25H, and a second mutation of L92H.

40 . The polypeptide of claim 39 , comprising a third mutation selected from the group consisting of C115S, C115A, K121R, K121Y, K121H, K121D, K121L, K121T, K121A, K121N, E123T, M126F, M126V, M126I, M126Q, M126N, K164S, L166N, L166M, L166I, L166T, L166A, I170N, I170Q, I170V, I170T, I170A, D172N, M194A, M194L, I195G, I195A, I195V, and K215N.

41 - 44 . (canceled)

45 . A method according to reaction Scheme B:

wherein
R 1 is (C 1 -C 6 )alkyl;
R 2 is —CF 2 H; and
the second enzyme is selected from the group consisting of Alcalase® 2.4 L, Esperase® 8.0 L, and Savinase® 16.0 L.

46 . (canceled)

47 . The method of claim 45 , wherein the loading of the second enzyme is about 50 wt % to about 200 wt % as compared to the starting material in Scheme B.

48 . The method of claim 45 , wherein the second solvent comprises an aqueous buffer.

49 . (canceled)

50 . The method of claim 48 , wherein the second solvent further comprises an organic solvent such as acetone.

51 - 53 . (canceled)

Assignees

• Abbvie Inc

Inventors

• Abrahamson Michael J
• Chemburkar Sanjay R
• Kielbus Angelica B
• Cink Russell D

Classifications

• C12N9/18 Primary

  Carboxylic ester hydrolases {(3.1.1)} · CPC title

• C12Y304/21014

  Microbial serine proteases (3.4.21.14) (C12Y304/21062 - C12Y304/21067 takes precedence) · CPC title

• C12Y301/01001

  Carboxylesterase (3.1.1.1) · CPC title

• C12P7/62 Primary

  Carboxylic acid esters · CPC title

• C12P41/005

  by esterification of carboxylic acid groups in the enantiomers or the inverse reaction · CPC title