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Biocatalytic desymmetrization of 3-substituted glutaronitriles by nitrilases. A convenient chemoenzymatic access to optically active (S)-Pregabalin and (R)-Baclofen

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Abstract

Desymmetrization of prochiral 3-substituted glutaronitriles offers a new approach to access (S)-Pregabalin and (R)-Baclofen. A number of nitrilases from diverse sources were screened with 3-isobutylglutaronitriles (1a) or 3-(4′-chlorophenyl)glutaronitriles (1b) as the substrate. Some nitrilases were found to catalyze the desymmetric hydrolysis of 1a and 1b to form optically active 3-(cyanomethyl)-5-methylhexanoic acid (2a) and 3-(4′-chlorophenyl)-4-cyanobutanoic acid (2b) with high enantiomeric excesse (ee), respectively. This cannot be achieved using traditional chemical hydrolysis. Among them, AtNIT3 generated (R)-2b, whereas BjNIT6402 and HsNIT produced the opposite (S)-enantiomer with high conversions and ee values. Not only the nitrilases showed different activities and stereoselectivities toward these 3-substituted glutaronitriles, the 3-substituent of the substrates also exerted great effect on the enzyme activity and stereoselectivity. (S)-2a and (S)-2b were prepared with high yields and ee values using BjNIT6402 and HsNIT as the biocatalysts, respectively. A straightforward Curtius rearrangement of (S)-2a and (S)-2b, followed by the acidic hydrolysis, afforded (S)-Pregabalin and (R)-Baclofen. This offers a new platform methodology for the synthesis of optically active β-substituted γ-amino acids of pharmaceutical importance.

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References

  1. Wall GM, Baker JK. Metabolism of 3-(p-chlorophenyl) pyrrolidine. Structural effects in conversion of a prototype γ-aminobutyric acid prodrug to lactam and γ-aminobutyric acid type metabolites. J Med Chem, 1989, 32: 1340–1348

    Article  CAS  Google Scholar 

  2. Ordóñez M, Cativiela C. Stereoselective synthesis of γ-amino acids. Tetrahedron: Asymmetry, 2007, 18: 3–99

    Article  Google Scholar 

  3. Garbutt JC. Emerging pharmacologic treatments for alcohol dependence. J Clin Psychiat, 2006, 67: 35–40

    CAS  Google Scholar 

  4. Sofuoglu M, Lappalainen J. GABAergic agents for the treatment of nicotine dependence. In: George TP. Medication Treatments for Nicotine Dependence. Boca Raton: Taylor & Francis Group, 2007

    Google Scholar 

  5. Liu JM, Wang X, Ge ZM, Sun Q, Cheng TM, Li RT. Solvent-free organocatalytic Michael addition of diethyl malonate to nitroalkenes: the practical synthesis of Pregabalin and γ-nitrobutyric acid derivatives. Tetrahedron, 2011, 67: 636–640

    Article  CAS  Google Scholar 

  6. Shao C, Yu HJ, Wu NY, Tian P, Wang R, Feng CG, Lin GQ. Asymmetric synthesis of β-substituted γ-lactams via rhodium/diene-catalyzed 1,4-additions: application to the synthesis of (R)-Baclofen and (R)-Rolipram. Org Lett, 2011, 13: 788–791

    Article  CAS  Google Scholar 

  7. Yu HJ, Shao C, Cui Z, Feng CG, Lin GQ. Highly enantioselective alkenylation of cyclic α,β-unsaturated carbonyl compounds as catalyzed by a rhodium-diene complex: application to the synthesis of (S)-Pregabalin and (−)-α-Kainic acid. Chem Eur J, 2012, 18: 13274–13278

    Article  CAS  Google Scholar 

  8. Yang XF, Ding CH, Li XH, Huang JQ, Hou XL, Dai LX, Wang PJ. Regio- and enantioselective palladium-catalyzed allylic alkylation of nitromethane with monosubstituted allyl substrates: synthesis of (R)-Rolipram and (R)-Baclofen. J Org Chem, 2012, 77: 8980–8985

    Article  CAS  Google Scholar 

  9. Baran R, Veverkova E, Skvorcova A, Sebesta R. Enantioselective Michael addition of 1,3-dicarbonyl compounds to a nitroalkene catalyzed by chiral squaramides: a key step in the synthesis of Pregabalin. Org Biomol Chem, 2013, 11: 7705–7711

    Article  CAS  Google Scholar 

  10. Palomo MJ, Cabrera Z. Enzymatic desymmetrization of prochiral molecules. Curr Org Synth, 2012, 9: 791–805

    Article  CAS  Google Scholar 

  11. Garcia-Urdiales E, Alfonso I, Gotor V. Update 1 of: enantioselective enzymatic desymmetrizations in organic synthesis. Chem Rev, 2011, 111: PR110–PR180

    Article  Google Scholar 

  12. Bayer S, Birkemeyer C, Ballschmiter M. A nitrilase from a metagenomic library acts regioselectively on aliphatic dinitriles. Appl Microbiol Biot, 2011, 89: 91–98

    Article  CAS  Google Scholar 

  13. Effenberger F, Osswald S. Selective hydrolysis of aliphatic dinitriles to monocarboxylic acids by a nitrilase from Arabidopsis thaliana. Synthesis-Stuttgart, 2001: 1866–1872

    Google Scholar 

  14. Yoshida T, Mitsukura K, Mizutani T, Nakashima R, Shimizu Y, Kawabata H, Nagasawa T. Enantioselective synthesis of (S)-2-cyano-2-methylpentanoic acid by nitrilase. Biotechnol Lett, 2013, 35: 685–688

    Article  CAS  Google Scholar 

  15. Zhu DM, Mukherjee C, Biehl ER, Hua L. Nitrilase-catalyzed selec tive hydrolysis of dinitriles and green access to the cyanocarboxylic acids of pharmaceutical importance. Adv Synth Catal, 2007, 349: 1667–1670

    Article  CAS  Google Scholar 

  16. Mukherjee C, Zhu DM, Biehl ER, Hua L. Exploring the synthetic applicability of a cyanobacterium nitrilase as catalyst for nitrile hydrolysis. Eur J Org Chem, 2006, 2006: 5238–5242

    Article  Google Scholar 

  17. Vorwerk S, Biernacki S, Hillebrand H, Janzik I, Muller A, Weiler EW, Piotrowski M. Enzymatic characterization of the recombinant Arabidopsis thaliana nitrilase subfamily encoded by the NIT2/NIT1/NIT3-gene cluster. Planta, 2001, 212: 508–516

    Article  CAS  Google Scholar 

  18. Liu ZQ, Dong LZ, Cheng F, Xue YP, Wang YS, Ding JN, Zheng YG, Shen YC. Gene cloning, expression, and characterization of a nitrilase from Alcaligenes faecalis ZJUTB10. J Agr Food Chem, 2011, 59: 11560–11570

    Article  CAS  Google Scholar 

  19. Desantis G, Wong K, Farwell B, Chatman K, Zhu ZL, Tomlinson G, Huang HJ, Tan XQ, Bibbs L, Chen P, Kretz K, Burk MJ. Creation of a productive, highly enantioselective nitrilase through gene site saturation mutagenesis (GSSM). J Am Chem Soc, 2003, 125: 11476–11477

    Article  CAS  Google Scholar 

  20. Zhu DM, Mukherjee C, Yang Y, Rios BE, Gallagher DT, Smith NN, Biehl ER, Hua L. A new nitrilase from Bradyrhizobium japonicum USDA 110. Gene cloning, biochemical characterization and substrate specificity. J Biotechnol, 2008, 133: 327–333

    Article  CAS  Google Scholar 

  21. Zhu DM, Mukherjee C, Biehl ER, Hua L. Discovery of a mandelonitrile hydrolase from Bradyrhizobium japonicum USDA110 by rational genome mining. J Biotechnol, 2007, 129: 645–650

    Article  CAS  Google Scholar 

  22. Mukherjee C, Zhu DM, Biehl ER, Parmar RR, Hua L. Enzymatic nitrile hydrolysis catalyzed by nitrilase ZmNIT2 from maize. An unprecedented beta-hydroxy functionality enhanced amide formation. Tetrahedron, 2006, 62: 6150–6154

    Article  CAS  Google Scholar 

  23. Fawcett JK, Scott JE. A rapid and precise method for the determination of urea. J Clin Pathol, 1960, 13: 156–159

    Article  CAS  Google Scholar 

  24. Hameršak Z, Stipetić I, Avdagić A. An efficient synthesis of (S)-3-aminomethyl-5-methylhexanoic acid (Pregabalin) via quininemediated desymmetrization of cyclic anhydride. Tetrahedron: Asymmetry, 2007, 18: 1481–1485

    Article  Google Scholar 

  25. Wang MX, Liu CS, Li JS. Enzymatic desymmetrization of 3-alkyl- and 3-arylglutaronitriles, a simple and convenient approach to optically active 4-amino-3-phenylbutanoic acids. Tetrahedron: Asymmetry, 2002, 12: 3367–3373

    Article  CAS  Google Scholar 

  26. Martínková L, Vejvoda V, Kaplan O, Křen V, Bezouška K, Cantarella M. Nitrilases from filamentous fungi. In: Fessner WD, Anthonsen T. Modern Biocatalysis: Stereoselective and Environmentally Friendly Reactions. Weinheim: Wiley-VCH, 2009

    Google Scholar 

  27. Raczynska JE, Vorgias CE, Antranikian G, Rypniewski W. Crystallographic analysis of a thermoactive nitrilase. J Struct Biol, 2011, 173: 294–302

    Article  CAS  Google Scholar 

  28. Mueller P, Egorova K, Vorgias CE, Boutou E, Trauthwein H, Verseck S, Antranikian G. Cloning, overexpression, and characterization of a thermoactive nitrilase from the hyperthermophilic archaeon Pyrococcus abyssi. Protein Expres Purif, 2006, 47: 672–681

    Article  CAS  Google Scholar 

  29. Novo C, Tata R, Clemente A, Brown PR. Pseudomonas aeruginosa aliphatic amidase is related to the nitrilase/cyanide hydratase enzyme family and Cys166 is predicted to be the active site nucleophile of the catalytic mechanism. FEBS Lett, 1995, 367: 275–279

    Article  CAS  Google Scholar 

  30. Stevenson DE, Feng R, Dumas F, Groleau D, Mihoc A, Storer AC. Mechanistic and structural studies on Rhodococcus ATCC 39484 nitrilase. Biotechnol Appl Bioc, 1992, 15: 283–302

    CAS  Google Scholar 

  31. Stevenson DE, Feng R, Storer AC. Detection of covalent enzyme-substrate complexes of nitrilase by ion-spray mass spectroscopy. FEBS Lett, 1990, 277: 112–114

    Article  CAS  Google Scholar 

  32. **e ZY, Feng JL, Garcia E, Bernett M, Yazbeck D, Tao JH. Cloning and optimization of a nitrilase for the synthesis of (3S)-3-cyano-5-methyl hexanoic acid. J Mol Catal B: Enzym, 2006, 41: 75–80

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. National Engineering Laboratory for Industrial Enzymes and Tian** Engineering Center for Biocatalytic Technology, Tian** Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tian**, 300308, China

    YiTao Duan, PeiYuan Yao, Jie Ren, Chao Han, Qian Li, **g Yuan, **Hui Feng, QiaQing Wu & DunMing Zhu

  2. College of Biotechnology, Tian** University of Science and Technology, Tian**, 300457, China

    Chao Han

Authors
  1. YiTao Duan
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  2. PeiYuan Yao
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  3. Jie Ren
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  4. Chao Han
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  5. Qian Li
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  6. **g Yuan
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  7. **Hui Feng
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  8. QiaQing Wu
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  9. DunMing Zhu
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Corresponding authors

Correspondence to QiaQing Wu or DunMing Zhu.

Additional information

Dedicated to Professor Qian Changtao on the occasion of his 80th birthday.

These authors contributed equally to this work

WU QiaQing received his BSc, MSc in Biochemistry and PhD in Genetics from Sichuan University in 1986, 1992 and 2004, respectively. He is a Professor in Tian** Institute of Industrial Biotechnology, Chinese Academy of Sciences. Before joining TIB, he was Deputy Director in the department of genetic engineering drugs, Chengdu Di-Ao pharmaceutical group Co., Ltd. His current research interests are focused on discovery, improvement and applications of industrial enzymes.

ZHU DunMing obtained a BSc from University of Science and Technology of China in 1987, and his MSc and PhD from Shanghai Institute of Organic Chemistry in 1990 and 1993, respectively, under the supervision of Professor Changtao Qian. He is a Professor in Tian** Institute of Industrial Biotechnology, Chinese Academy of Sciences. He was honored by “Hundred Talents Program” of Chinese Academy of Sciences in 2008, and “Thousand Talents Program” of Tian** City in 2009. His research interests range from discovery of novel industrial enzymes to understanding of biocatalytic reaction mechanisms, and the integration of biocatalysis into complex organic synthesis.

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Duan, Y., Yao, P., Ren, J. et al. Biocatalytic desymmetrization of 3-substituted glutaronitriles by nitrilases. A convenient chemoenzymatic access to optically active (S)-Pregabalin and (R)-Baclofen. Sci. China Chem. 57, 1164–1171 (2014). https://doi.org/10.1007/s11426-014-5139-2

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