Antimitotic natural products combretastatin A-4 and combretastatin A-2: studies on the mechanism of their inhibition of the binding of colchicine to tubulin.
Combretastatin A-4 (CS-A4), 3,4,5-trimethoxy-3'-hydroxy-4'-methoxy-(Z)-stilbene, and combretastatin A-2 (CS-A2), 3,4-(methylenedioxy)-5-methoxy-3'-hydroxy-4'-methoxy-(Z)-stilbene, are structurally simple natural products isolated from the South African tree Combretum caffrum. They inhibit mitosis and microtubule assembly and are competitive inhibitors of the binding of colchicine to tubulin [Lin et al. (1988) Mol. Pharmacol. 34, 200-208]. In contrast to colchicine, drug effects on tubulin were not enhanced by preincubating CS-A4 or CS-A2 with the protein. The mechanism of their binding to tubulin was examined indirectly by evaluating their effects on the binding of radiolabeled colchicine to the protein. These studies demonstrated rapid binding of both compounds to tubulin even at 0 degrees C (binding was complete at the earliest times examined), in contrast to the relatively slow and temperature-dependent binding of colchicine. Although the binding of the C. caffrum compounds to tubulin was quite tight, permitting ready isolation of near-stoichiometric amounts of drug-tubulin complex even in the absence of free drug, both CS-A4 and CS-A2 dissociated rapidly from tubulin in the presence of high concentrations of radiolabeled colchicine. Apparent rate constants for drug dissociation from tubulin at 37 degrees C were 3.2 x 10(-3) s-1 for CS-A4, 4.8 x 10(-3) s-1 for CS-A2, and 2.9 x 10(-5) s-1 for colchicine (half-lives of 3.6, 2.4, and 405 min, respectively). Thus, the effectiveness of the C. caffrum compounds as antimitotic agents appears to derive primarily from the rapidity of their binding to tubulin.(ABSTRACT TRUNCATED AT 250 WORDS)
Lin
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Division of Cancer Treatment, National Cancer Institute, Bethesda, Maryland 20892.
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Pettit
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Hamel
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D000818:Animals; D001632:Bibenzyls / Q000494:pharmacology*; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D003078:Colchicine / Q000378:metabolism* / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D046911:Macromolecular Substances; D008938:Mitosis / Q000187:drug effects; D008961:Models, Structural; D010936:Plant Extracts; D011034:Podophyllotoxin / Q000494:pharmacology; D011485:Protein Binding; D013267:Stilbenes / Q000494:pharmacology*; D014334:Tropolone / Q000494:pharmacology / Q000031:analogs & derivatives; D014404:Tubulin / Q000378:metabolism*
D001632:Bibenzyls; D046911:Macromolecular Substances; D010936:Plant Extracts; D013267:Stilbenes; D014404:Tubulin; C061751:combretastatin A-2; C030370:2-methoxy-5-(2',3',4'-trimethoxyphenyl)tropone; D014334:Tropolone; D006160:Guanosine Triphosphate; C058728:fosbretabulin; D011034:Podophyllotoxin; D003078:Colchicine