Affiliation:
1. Department of Medicine, University of California, San Francisco 94143, USA.
Abstract
An increase in the number of tuberculosis cases caused by multiple-drug-resistant strains of Mycobacterium tuberculosis has stimulated search for new antituberculous agents. Beta-lactam antibiotics, traditionally regarded as ineffective against tuberculosis, merit consideration. Four major penicillin-binding proteins (PBPs) with approximate molecular sizes of 94, 82, 52, and 37 kDa were detected by fluorography of [3H]penicillin-radiolabeled membrane proteins prepared from M. tuberculosis H37Ra. The presence of membrane-associated beta-lactamase precluded the use of membranes for assaying the binding affinities of beta-lactam antibiotics. Therefore, ampicillin affinity chromatography was used to purify these four PBPs from crude membranes in order to assay the binding affinities of beta-lactam antibiotics. Ampicillin, amoxicillin, and imipenem, beta-lactam antibiotics previously reported to be active in vitro against M. tuberculosis, bound to M. tuberculosis PBPs at therapeutically achievable concentrations. Binding of the 94-, 82-, and 52-kDa PBPs, but not the 37-kDa PBP, was associated with antibacterial activity, suggesting that these PBPs are the critical targets. Studies of mycobacterial cell wall permeability, which was assayed with a panel of reference cephalosporins and penicillins with different charge positivities, indicated that the rate of penetration of beta-lactam antibiotics to the target PBPs could not account for resistance. Resistance could be reversed with the beta-lactamase inhibitors clavulanate or sulbactam or could be circumvented by the use of a beta-lactamase-stable drug, imipenem, indicating that mycobacterial beta-lactamase, probably in conjunction with slow penetration, is a major determinant of M. tuberculosis resistance to beta-lactam antibiotics. These findings confirm in vitro data that M. tuberculosis is susceptible to some beta-lactam antibiotics. Further evaluation of these drugs for the treatment of tuberculosis in animal models and in clinical trials is warranted.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Pharmacology (medical),Pharmacology
Reference36 articles.
1. Pharmacokinetics of amoxicillin and clavulanic and administered alone and in combination;Adam D.;Antimicrob. Agents Chemother.,1982
2. Tuberculosis in the 1990s;Barnes P.;Ann. Intern. Med.,1993
3. Bryan L. and A. Godfrey. 1991. Beta-lactam antibiotics: mode of action and bacterial resistance p. 599-664. In V. Lorian (ed.) Antibiotics in laboratory medicine 3rd ed. The Williams & Wilkins Co. Baltimore.
4. Characterization of beta-lactamases;Bush K.;Antimicrob. Agents Chemother.,1989
5. Characterization of beta-lactamases: groups 1, 2a, 2b, and 2b~;Bush K.;Antimicrob. Agents Chemother.,1989
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