Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture
  • C12P41/006—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures
  • C12P41/007—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture by reactions involving C-N bonds, e.g. nitriles, amides, hydantoins, carbamates, lactames, transamination reactions, or keto group formation from racemic mixtures by reactions involving acyl derivatives of racemic amines
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
  • C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
  • C12P17/165—Heterorings having nitrogen atoms as the only ring heteroatoms

Definitions

• This invention provides an enzymatic process for preparing optically enriched intermediates useful in the preparation of substituted tricyclic compounds known as antihistamines and as inhibitors of famesyl protein transf erase (FPT).
• the process of this invention is useful in preparing intermediates useful in the preparation of FPT inhibitors disclosed, for example, in International Publication Number W095/10516, published April 20, 1995.
• the resolutions suffer from low reactivity and/or selectivity.
• This invention provides a highly selective process for preparing a substituted (6,11 -dihydro-5H-benzo[5,6]cyclohepta[1 ,2-b]-pyridin-11 - yl)piperidine compound of the formula (+)-I
• R, R 1 , R 2 , R 3 and R 4 are independently selected from the group consisting of hydrogen, halo, C-i-C ⁇ alkyl, amino, -OCH3, -OCF3 and CF3, and the dotted line represents an optional double bond; comprising:
• the enzyme is a hydrolase and wherein the acylating agent is of the formula R 5 COOR 6 , wherein R 5 is C1-C-15 alkyl, halo methyl, aryl, benzyl or benzyloxy, R 6 is alkyl, C1-C6 alkenyl, -COR 7 , trifluoroethyl, -CH 2 CH(OCOR 7 )CH2 ⁇ COR 7 , halo methyl or benzyl, and R 7 is C ⁇ -C ⁇ 5 alkyl; and
• step (c) optionally converting an undesired isomer from step (a) wherein a double bond is present to the racemate by heating, and resubjecting the racemate to enzymatic acylation and hydrolysis; or
• Preferred compounds of formula (+)-I made by this process are those wherein R 3 is not hydrogen. Also preferred are compounds wherein R is halo. Still another group of preferred compounds is that wherein R 1 is hydrogen and R, R 2 , and R 3 are selected form the group consisting of halo. Halo is preferably Cl or Br.
• halo means fluoro, chloro, bromo and iodo, with chloro and bromo being preferred.
• aryl means phenyl, naphthyl, substituted phenyl or substituted naphthyl, wherein “substituted” means 1-3 substituents indpendently selected form the group consisting of C-I-C6 alkyl, C-i-C ⁇ alkoxy, halo, NO2 and halo methyl.
• acylating enzymes may have opposing selectivity, and therefore may involve either direct or subtractive resolution. That is, some enzymes may acylate the desired isomer, requiring separation of the isomers, followed by hydrolysis to obtain the desired product (i.e., direct resolution, as claimed in step (1)), while others may acylate the undesired isomer, requiring only separation of the isomers (no hydrolysis) to obtain the desired isomer (i.e., subtractive resolution, as claimed in step (2)).
• Preferred enzymes are Toyobo LIP-300/301 , Altus Chiro CLECTM PC, Boehringer Mannheim Lipase, Novo SP435 and Novozym 435.
• Acylating agents of formula R 5 COOR 6 are commercially available or can be prepared by known methods.
• Preferred acylating agents are trifluoroethyl acetate (TFEOAc), trifluoroethyl butyrate (TFEOBu), trifluoroethyl isobutyrate (TFEOiBu), trifluoroethyl benzoate (TFEBenz), triacetin and tributyrin.
• the enzymatic acylation may be carried out in a solvent such as an alkyl acetate such as methyl acetate (MeOAc) or isopropyl acetate, t-butyl methyl ether (TBME), tetrahydrofuran (THF), acetone, acetonitrile, t-amyl alcohol, t-butyl alcohol, pyridine or dioxane.
• a solvent such as an alkyl acetate such as methyl acetate (MeOAc) or isopropyl acetate, t-butyl methyl ether (TBME), tetrahydrofuran (THF), acetone, acetonitrile, t-amyl alcohol, t-butyl alcohol, pyridine or dioxane.
• a solvent such as an alkyl acetate such as methyl acetate (MeOAc) or isopropyl acetate, t-butyl
• the reaction is carried out in a temperature range of 0 to 50°C, preferably at 25 to 30°C (e.g., ambient temperature).
• the reaction time ranges from 18 to 48 hours, with 24 hours being preferred.
• the enzyme is added at a ratio of about 1 : 2 times the amount of the starting material , preferably about 2 times the amount.
• the acylating agent is present at about 2 to 10 times the starting material, preferably about 3 times the amount of starting material when the enzyme is present at 2 times the amount of the starting material.
• the hydrolysis is carried out using standard procedures well known in the art. For example, the acylated compound is refluxed with an acid such as H2SO4. The desired isomer is then recovered by precipitating out by adding a base such as NaOH.
• the reaction is preferably carried out under anhydrous conditions.
• the solvent, or acylating agent when used as the solvent can be anhydrous, or the solution of the starting material in the solvent, or acylating agent when used as the solvent, can be dried by azeotropic distillation before the enzyme is added.
• the enzyme should be dried under vacuum before adding to the solution, preferably to ⁇ 700 ppm water.
• the undesired (-) isomer can be recovered from enzymatic isobutyrylation of racemic II. Heating (-)-ll in diphenyl ether or diethylene glycol dibutyl ether (5-15:1 , v:v) at 200-260°C, preferably 210°C, for 0.5- 26 hours results in complete racemization to racemic II which can be recovered in 77-95% yield with 93-99% purity.
• the recovered racemic II undergoes the enzymatic isobutyrylation under the same conditions as above.
• Previous methods used to resolve isomers to obtain compounds of formula I involved the resolution of a compound of formula ( ⁇ )-IIa by chiral chromatography or chemical resolution using stiochiometric amounts of a resolving agent.
• the process claimed herein uses a biocatalyst to effect the resolution, the biocatalyst being reusable up to 15 times.
• compounds lib are chiral atropisomers at room temperature due to restricted rotation about the double bond.
• the isomers can be racemized at high temperatures.
• the undesired (-) isomer can be isolated, racemized at 200-260°C, preferably at 210°C, and then subjected to a further enzymatic acylation to increase throughput of ( ⁇ )-IIIa.
• the products of this process are intermediates useful in the preparation of tricyclic compounds useful as famesyl protein trasnsferase inhibitors such as those disclosed in International Publication Number WO95/10516, published April 20, 1995.
• ee s is the enantiomeric excess of the unreacted starting material
• eep is the enantiomeric excess of the product
• c is the conversion (ee s /(ee s + ee p ))
• E is the Enantiomeric Ratio: (ln[(1-ee s ) (1-c)/ln[(1+ee s ) (1-c)] or ln[1-c(1+ee p )]/ln[1-c(1+ eep)]);
• Ac is acetyl
• OAc is acetate
• Me is methyl
• Et is ethyl
• Pr is propyl
• TFE is trifluoroethyl.
• Runs 2,3, 5-11 ( ⁇ )-IIc, 10 mg; Trifluoroethyl Acetate, 20 equiv; TBME, 1.0 mL; RT, 250 ⁇ m.
• Runs 1 ,4 ( ⁇ )-IIc, 10 mg; CaC ⁇ 3, 30-40 mg; MeOAc, 1.0 mL (as solvent and acylating agent); RT; 250 rpm.
• Run12 (+)-IIc, 12 mg; Trifluoroethyl acetate, 5 equiv.; TBME, 1.0 mL; RT, 250 rpm B.
• Runs 1-9 ( ⁇ )-IIc (50 mg, 50mM), Toyobo LIP-300 (50-55 mg), TFEOiBu (5 eq., except Runs 1-3 which used solvent as acylating agent), Solvent (2.0 mL), 4A Sieves (47-59 mg), 250 rpm, RT, 22.25 h.
• Runs 1 ,3-7,9, 13-15 ( ⁇ )-IId, 7-11 mg; TFEOAc, 50 mL, 14-25 equiv.; SP435.8-13 mg; Solvent, 2.0 mL, RT, 250 ⁇ m.
• Runs 2, 8, 10-12 ( ⁇ )-IId, 12-15 mg; TFEOAc, 100 mL, 25-31 equiv.(except Run 2 which used solvent as the acylating agent); SP 435, 4-7 mg; Solvent, 2.0 mL; 250 rpm.
• Runs 1 , 4-9, 11-17, 19 ( ⁇ )-IId ,4.9 mg; TBME or neat acylating agent, 1.0 mL; SP 435, 6.2-10.8 mg; 250 rpm; RT.
• Run 1 1 , 5 mg; LIP-301 , 10 mg; Trifluoroethyl isobutyrate, 10 equivs.; TBME, 1.0 mL; 200 rpm; RT
• Run 2 2, 5.4 mg; LIP-301 , 16.6 mg; Trifluoroethyl isobutyrate, 20 equivs.; TBME, 1.0 mL; 200 rpm, RT.
• Run 3 3, 0.2 g; LIP-301 , 0.4 g; Trifluoroethyl isobutyrate, 10 equivs.; TBME, 4 mL;
• TBME (600 mL) was stirred at ambient temperature for 1 h, then filtered to remove a small amount of insoluble material. The solution was dried by azeotropic distillation; after 200 mL was distilled, a further 200 mL of TBME was added to the reaction mixture. After a total of 400 mL had been distilled, the moisture content (Karl-Fischer) of the solution was 214 ppm. Toyobo LIP-300 (40 g; 1282 ppm water) was added to the reaction mixture and stirred for 0.5 h; moisture content at this stage was 250 ppm.
• Trifluoroethyl isobutyrate (19.1 mL, 3 equivs.) was added and the mixture was stirred at ambient temperature. The reaction was terminated after 24 h. The enzyme was removed by filtration and washed with TBME (100 mL).
• the reaction was stirred at room temperature under N 2 for 24 h.
• the enzyme was then removed by filtration and washed with TBME (0.9 L).
• the combined filtrates were extracted sequentially with three portions of 0.5 M H2SO4 (450 mL, 225 mL and 225 mL). These combined acidic extracts contained the unreacted (-)-llc.
• the organic layer was then extracted with two portions of 6M H2SO4 (135 mL and 135 mL). These combined acidic extracts contained the product isobutyramide (+)-lllb.
• (+)-lc The combined 6M H2SO4 extracts were heated at reflux for 14 .5 h, then cooled to room temperature. The reaction mixture was then added slowly to a cold, vigorously stirred mixture of NH4OH (900 mL) and CH3CN (270 mL). The solid which precipitated was filtered, washed with water and dried (40.5 g, 43.5%; 0.960 ee).
• Isolation of (-)-llb The combined 0.5 M H2SO4 extracts were added slowly to a cold, vigorously stirred mixture of NH4OH (450 mL) and CH3CN (270 mL).
• the combined filtrate was extracted with three portions of 0.5 M H2SO4 (150 mL, 75 mL and 75 mL) to remove the unreacted (-)-llc.
• the organic layer was then extracted with two portions of 6M H2SO4 (45 mL and 45 mL) which were combined and refluxed for 16 h.
• the cooled reaction mixture was then added slowly to a vigorously stirred, cold mixture of NH4OH (300 mL) and CH3CN (90 mL).
• the precipitated (+)-lc was filtered, washed with water and dried: (13 g, 43%; 0.986 ee).

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Cited By (3)

• 1998
  • 1998-06-15 ZA ZA985217A patent/ZA985217B/xx unknown
  • 1998-06-15 IN IN1292MA1998 patent/IN187248B/en unknown
  • 1998-06-15 EP EP98928889A patent/EP1002123A1/en not_active Withdrawn
  • 1998-06-15 WO PCT/US1998/011501 patent/WO1998058073A1/en not_active Application Discontinuation
  • 1998-06-15 MY MYPI98002658A patent/MY133532A/en unknown
  • 1998-06-15 JP JP50449699A patent/JP3421355B2/ja not_active Expired - Fee Related
  • 1998-06-15 AU AU80583/98A patent/AU8058398A/en not_active Abandoned
  • 1998-06-15 CA CA002293708A patent/CA2293708C/en not_active Expired - Fee Related
  • 1998-06-16 AR ARP980102861A patent/AR015892A1/es unknown

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