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DOI: 10.1039/A700998D (Paper) Reference Section for: J. Chem. Soc., Perkin Trans. 2, 1997, 2439-2444

β-Lactamase-catalysed hydrolysis of cephalexin: evolution of the cephalosporoate intermediate

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Bartolomé Vilanova, Juan Frau, Josefa Donoso, Francisco Muñoz and Francisco García Blanco

Abstract

Hydrolysis of cephalexin catalysed by P99 β-lactamase from Enterobacter cloacae has been studied at pD 6.4 and 8.0. The enzymatic hydrolysis product is the cephalosporoate intermediate, which in aqueous media undergoes tautomerization of the double bond in the dihydrothiazine ring from position 3–4 to 4–5, with the consequent uptake of a proton at C(3). This latter process is governed by steric constraints. NMR results show that the cephalosporoate compound bears the aromatic group, present in the side chain at C(7), folded over the dihydrothiazine ring. Under these conditions the proton is chiefly taken up by the α face of the dihydrothiazine ring. Subsequently, the six-membered ring can be subjected to several reactions according to the pH.

References

  1. S. G. Waley, in The Chemistry of β-lactams, Blackie, Glasgow, 1992, pp. 198–228 Search PubMed.
  2. S. G. Waley, Sci. Prog. (Oxford), 1988, 72, 579 Search PubMed.
  3. P. Ledent, X. Raquet, B. Joris, L. Van Beeumen and J.-M. Frère, Biochem. J., 1993, 292, 555 CAS.
  4. R. F. Pratt and W. S. Faraci, J. Am. Chem. Soc., 1986, 108, 5328 CrossRef CAS.
  5. W. S. Faraci and R. F. Pratt, Biochemistry, 1985, 24, 903 CrossRef CAS.
  6. J. M. T. Hamilton-Miller, G. G. F. Newton and E. P. Abraham, Biochem. J., 1970, 116, 371 CAS.
  7. J. M. T. Hamilton-Miller, E. Richards and E. P. Abraham, Biochem. J., 1970, 116, 385 CAS.
  8. S. Mobashery and M. Johnston, J. Biol. Chem., 1986, 261, 7879 CAS.
  9. R. N. Jones and D. A. Preston, Postgrad. Med. J., 1983, 59(Suppl. 5), 9 CAS.
  10. R. B. Kammer, in Chemistry and Biology of β-lactam Antibiotics, Academic Press, New York, 1982, pp. 287–300 Search PubMed.
  11. H. Bundgaard, Arch. Pharm. Chem. Sci. Ed. 4, 1976, 25 Search PubMed.
  12. H. Bundgaard, Arch. Pharm. Chem. Sci. Ed. 5, 1977, 149 Search PubMed.
  13. B. Vilanova, F. Muñoz, J. Donoso and F. García Blanco, Helv. Chim. Acta, 1993, 76, 2789 CrossRef CAS.
  14. B. Vilanova, F. Muñoz, J. Donoso, J. Frau and F. García Blanco, J. Pharm. Sci., 1994, 83, 322 CAS.
  15. G. B. Young and T. L. James, J. Am. Chem. Soc., 1984, 106, 7986 CrossRef CAS.
  16. C. García-Echevarría, G. Siligardi, P. Mascagni, W. Gibbons, E. Giralt and M. Pons, Biopolymers, 1991, 31, 835 CrossRef.
  17. S. Macura, B. T. Farmer II and L. R. Brown, J. Magn. Reson., 1986, 70, 493 CAS.
  18. G. Esposito, J. A. Carver, J. Boyd and I. D. Campbell, Biochemistry, 1987, 26, 1043 CrossRef CAS.
  19. D. J. States, R. A. Haberkorn and B. J. Ruben, J. Magn. Reson., 1982, 48, 286 CAS.
  20. G. Bodenhausen and R. R. Ernst, J. Am. Chem. Soc., 1982, 104, 1304 CrossRef CAS.
  21. M. I. Page, B. Vilanova and N. J. Layland, J. Am. Chem. Soc., 1995, 117, 12 092 CrossRef CAS.
  22. A. I. Cohen, P. T. Funke and M. S. Puar, J. Pharm. Sci., 1973, 62, 1559 CAS.
  23. A. Dinner, J. Med. Chem., 1977, 20, 963 CrossRef CAS.
  24. S. W. Baertschi, D. E. Dorman, J. L. Occolowitz, L. A. Spangle, M. W. Collins, M. E. Wildfeuer and L. J. Lorenz, J. Pharm. Sci., 1993, 82, 622 CAS.
  25. B. Vilanova, J. Donoso, F. Muñoz and F. García-Blanco, Helv. Chim. Acta, 1996, 79, 1793 CrossRef CAS.
  26. P. Salomaa, L. L. Schaleger and F. A. Long, J. Am. Chem. Soc., 1964, 86, 1 CrossRef CAS.
  27. D. M. Doddrell, D. T. Pegg and M. R. Bendall, J. Magn. Reson., 1982, 48, 323.
  28. J. J. P. Stewart, J. Comput. Chem., 1989, 10, 209 CrossRef CAS.
  29. J. J. P. Stewart, J. Comput. Chem., 1989, 10, 221 CrossRef CAS.
  30. J. J. P. Stewart, J. Comput. Chem., 1991, 12, 320 CrossRef CAS.
  31. M. J. S. Dewar, E. G. Zoebisch, E. F. Healy and J. J. P. Stewart, J. Am. Chem. Soc., 1985, 107, 3902 CrossRef.
  32. ‘AMPAC 5.0, 1994 Semichem’, 7128 Summit, Shawnee, KS 66 216.
  33. J. Frau, J. Donoso, F. Muñoz and García Blanco, J. Mol. Struct. (THEOCHEM), 1991, 251, 205 CrossRef.
  34. S. K. Branch, A. F. Casy, A. Lipczynski and E. M. A. Ominde, Magn. Reson. Chem., 1986, 24, 465.