ANALOGS OF CHLORAMPHENICOL AS MECHANISM-BASED INACTIVATORS OF RAT LIVER CYTOCHROMES P-450
AuthorMILLER, NATALIE ELIZABETH.
Xenobiotics -- Metabolic detoxification.
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PublisherThe University of Arizona.
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AbstractThe cytochrome P-450 dependent monooxygenase system plays a key role in the bioactivation and detoxication of xenobiotics. Isozyme-specific inhibitors of cytochrome P-450 may be useful in elucidating the role of particular isozymes in xenobiotic metabolism or in suppressing the bioactivation of xenobiotics and enhancing detoxication. The antibiotic chloramphenicol is a selective mechanism-based inactivator of rat liver cytochromes P-450, inactivating 6 of the 12 isozymes monitored, including the major phenobarbital-inducible isozyme PB-B. Analogs of chloramphenicol have been tested to determine the importance of various functional groups in regulating the effectiveness and isozyme selectivity of chloramphenicol as a mechanism-based inactivator of cytochromes P-450. This information will aid in the design of more effective and isozyme specific mechanism-based inactivators. The dihalomethyl group and the propanediol moiety were found to be important in determining the efficacy of inactivation and the ability to inactivate the enzyme by virtue of the modification of the protein as opposed to the modification of the heme moiety. The propanediol side chain also plays a role in the isozyme selectivity. Unlike chloramphenicol, N (2-p-nitrophenethyl)dichloroacetamide (pNO₂DCA), which contains an ethyl group in place of the propanediol side chain of chloramphenicol, is an effective inactivator of BNF-B, the major beta-naphthoflavone-inducible isozyme, as well as PB-B, in vitro and in vivo. Alkaline hydrolysis and enzymatic digestion of the covalently modified isozymes has shown that chloramphenicol and pNO₂DCA are both metabolized by cytochromes P-450 to oxamyl chlorides which bind to lysine and other amino acid residues of the enzyme. However, the mechanism by which pNO₂DCA inactivates BNF-B differs significantly from that by which chloramphenicol inactivates PB-B, although both involve an impairment of the transfer of electrons from NADPH-cytochrome P-450 reductase, suggesting that there are differences in the active sites of these two isozymes.
Degree ProgramPharmacology & Toxicology