Penicillin acylase-catalyzed synthesis of N-bromoacetyl-7-aminocephalosporanic acid, the key intermediate for the production of cefathiamidine

Abstract

Background

Enzymatic approaches have become promising alternatives to chemical methods for the production of semi-synthetic β-lactam antibiotics. In this work, enzymatic synthesis of N-bromoacetyl-7-aminocephalosporanic acid (N-bromoacetyl-7-ACA), the key intermediate for the production of cefathiamidine, was reported for the first time.

Results

Of the immobilized penicillin acylases (PAs) tested, PGA-750 was the best biocatalyst. Optimization of the biocatalytic process was conducted. The optimal acyl donor, molar ratio of acyl donor to 7-ACA, pH, temperature, 7-ACA concentration, and enzyme dosage were methyl bromoacetate, 3, 7.5, 20 °C, 50 mmol/L and 4 U/mL, respectively. Under the optimal conditions, enzymatic N-acylation of 7-ACA with methyl bromoacetate afforded the desired product with the yield of 85% in 2 h, where the synthesis/hydrolysis (S/H) ratio was approximately 1.5. The immobilized enzyme PGA-750 exhibited good operational stability, and the relative yields of approximately 90% and 63% were achieved, respectively, when it was reused in 7th and 11th batch.

Conclusions

An enzymatic approach to N-bromoacetyl-7-ACA, the key intermediate for the industrial production of cefathiamidine, has been developed successfully in a fully aqueous medium. The present work may open up a novel opportunity for the production of cefathiamidine through a simple and green process.

Keywords

7-Aminocephalosporanic acid / Enzyme catalysis / Cephalosporins / Penicillin acylase / Semi-synthetic antibiotics

References

[1] Aguirre C, Opazo P, Venegas M, Riveros R, Illanes A. Low temperature effect on production of ampicillin and cephalexin in ethylene glycol medium with immobilized penicillin acylase. Process Biochem, 2006, 41(9): 1924-1931.

[2] Arroyo M, de la Mata I, Acebal C, Castillon MP. Biotechnological applications of penicillin acylases: state-of-the-art. Appl Microbiol Biotechnol, 2003, 60(5): 507-514.

[3] Bruggink A, Roos EC, de Vroom E. Penicillin acylase in the industrial production of β-lactam antibiotics. Org Proc Res Dev, 1998, 2(2): 128-133.

[4] Cao L, van Rantwijk F, Sheldon RA. Cross-linked enzyme aggregates: a simple and effective method for the immobilization of penicillin acylase. Org Lett, 2000, 2(10): 1361-1364.

[5] Chen H, Williams J. The killing effects of cefathiamidine or ampicillin alone and in combination with gentamicin against with gentamicin against enterococci. J Antimicrob Chemother, 1983, 12(1): 19-26.

[6] Dai ZY, Wang F, Zhang ZL, Zhang MF, Zhang JD, Zhang YY, Liu YK. Clinical and laboratory studies of a new cephalosporin derivative-cefathiamidine. Shanghai Med J, 1978, 1(9): 32-36.

[7] Ferreira ALO, Giordano RLC, Giordano RC. Nonconventional reactor for enzymatic synthesis of semi-synthetic β-lactam antibiotics. Ind Eng Chem Res, 2007, 46(23): 7695-7702.

[8] Fu HY, Tian MR. Improved preparation of cefathiamidine. Chin J Pharm, 2009, 40(12): 888-889.

[9] Illanes A, Wilson L, Corrotea O, Tavernini L, Zamorano F, Aguirre C. Synthesis of cephalexin with immobilized penicillin acylase at very high substrate concentrations in fully aqueous medium. J Mol Catal B Enzym, 2007, 47(1): 72-78.

[10] Li HQ. Effects of cefathiamidine on the treatment of children with acute bacterial infectious disease. Chin J Pediatr, 2003, 41(1): 54-55.

[11] Li L, Tang ZL, Yang SY, Zhang C, Hu XY, Zhang LP, Su W, Zhang LL. Interventional study on use time of prophylactic antibiotics for cesarean. Chin Hosp Pharm J, 2016, 36(10): 854-859.

[12] Liu DQ, Xu CS, Niu Y. Improved synthesis of cephathiamidine. Chin J Med Chem, 2001, 11(5): 293-294.

[13] Marešová H, Plačková M, Grulich M, Kyslík P. Current state and perspectives of penicillin G acylase-based biocatalyses. Appl Microbiol Biotechnol, 2014, 98(7): 2867-2879.

[14] Montes T, Grazu V, Manso I, Galan B, López-Gallego F, Gonzalez R, Hermoso JA, García JL, Guisán JM, Fernández-Lafuente R. Improved stabilization of genetically modified penicillin G acylase in the presence of organic cosolvents by co-immobilization of the enzyme with polyethyleneimine. Adv Synth Catal, 2007, 349(3): 459-464.

[15] Montoro-García S, Gil-Ortiz F, Navarro-Fernández J, Rubio V, García-Carmona F, Sánchez-Ferrer Á. Improved cross-linked enzyme aggregates for the production of desacetyl β-lactam antibiotics intermediates. Bioresour Technol, 2010, 101(1): 331-336.

[16] Ospina S, Barzana E, Ramírez OT, López-Munguía A. Effect of pH in the synthesis of ampicillin by penicillin acylase. Enzyme Microb Technol, 1996, 19(6): 462-469.

[17] Ribeiro MPA, Ferreira ALO, Giordano RLC, Giordano RC. Selectivity of the enzymatic synthesis of ampicillin by E. coli PGA in the presence of high concentrations of substrates. J Mol Catal B Enzym, 2005, 33(3–6): 81-86.

[18] Shewale JG, Kumar KK, Ambekar GR. Evaluation of determination of 6-aminopenicillanic acid by p-dimethyl aminobenzaldedyde. Biotechnol Tech, 1987, 1(1): 69-72.

[19] Srirangan K, Orr V, Akawi L, Westbrook A, Moo-Young M, Chou CP. Biotechnological advances on penicillin G acylase: pharmaceutical implications, unique expression mechanism and production strategies. Biotechnol Adv, 2013, 31(8): 1319-1332.

[20] Takimoto A, Takakura T, Tani H, Yagi S, Mitsushima K. Batch production of deacetyl 7-aminocephalosporanic acid by immobilized cephalosporin-C deacetylase. Appl Microbiol Biotechnol, 2004, 65(3): 263-267.

[21] Volpato G, Rodrigues RC, Fernandez-Lafuente R. Use of enzymes in the production of semi-synthetic penicillins and cephalosporins: drawbacks and perspectives. Curr Med Chem, 2010, 17(32): 3855-3873.

[22] Wang HL, Li LW. Improved synthesis process of cefathiamidine. Chin J Mod Appl Pharm, 2010, 27(2): 126-127.

[23] Wang WM, Su SH, Zhang SW, Yang B. A new semi-synthetic antibiotic-cefathiamidine (C-18). Chin J Pharm, 1978, 9(9): 1-4.

[24] Wegman MA, Janssen Michiel HA, van Rantwijk F, Sheldon Roger A. Towards biocatalytic synthesis of β-lactam antibiotics. Adv Synth Catal, 2001, 343(6–7): 559-576.

[25] Wegman MA, van Langen LM, van Rantwijk F, Sheldon RA. A two-step, one-pot enzymatic synthesis of cephalexin from d-phenylglycine nitrile. Biotechnol Bioeng, 2002, 79(3): 356-361.

[26] Wei D, Yang L, Song Q. Effect of temperature on the enzymatic synthesis of cefaclor with in situ product removal. J Mol Catal B Enzym, 2003, 26(1): 99-104.

[27] Yang L, Wei DZ. Enhanced enzymatic synthesis of a semi-synthetic cephalosporin, cefaclor, with in situ product removal. Biotechnol Lett, 2003, 25(14): 1195-1198.

[28] Yang GL, Zou T, Qu J, Yu L. Efficacy of cefathiamidine for the treatment of hemopathy with infection. Chin J New Drugs, 2006, 15(6): 470-472.

[29] Youshko MI, van Langen LM, de Vroom E, Moody HM, van Rantwijk F, Sheldon RA, Švedas VK. Penicillin acylase-catalyzed synthesis of ampicillin in “aqueous solution–precipitate” systems. High substrate concentration and supersaturation effect. J Mol Catal B Enzym, 2000, 10(5): 509-515.

[30] Youshko MI, Moody HM, Bukhanov AL, Boosten WH, Švedas VK. Penicillin acylase-catalyzed synthesis of β-lactam antibiotics in highly condensed aqueous systems: Beneficial impact of kinetic substrate supersaturation. Biotechnol Bioeng, 2004, 85(3): 323-329.

Funding

the National Natural Science Foundation of China(21676103)