膜蛋白和药物转运.ppt

P-GLYCOPROTEIN AND DRUG TRANSPORT Michael M. Gottesman Chief, Laboratory of Cell Biology, NCI Deputy Director for Intramural Research National Institutes of Health Mechanisms of resistance to anti-cancer drugs Drug Resistance in Cancer May affect multiple drugs used simultaneously: known as multidrug resistance (MDR) Affects all classes of drugs, including newly designed targetted drugs Just as oncogene targets have been catalogued, we need to enumerate all mechanisms of drug resistance in cancer to solve this problem and circumvent resistance Ultimate Goal: To use molecular analysis of human cancers to predict response to specific therapy To use this information to develop novel drugs to treat cancer To learn more about cellular pharmacology and pharmacokinetics of drugs Mechanisms of Resistance ATP-BINDING CASSETTE(N-terminal NBD of human Pgp) A C B Questions about the mechanism of action of P-glycoprotein How does P-glycoprotein recognize so many different substrates? What do the two ATP binding cassettes do? How is substrate binding linked to ATP hydrolysis? ATP sites in P-glycoprotein Both sites are essential; mutations in either site knock out transport function Sites work sequentially; only one site at a time binds and hydrolyzes ATP Stoichiometry of transport indicates that hydrolysis of two molecules of ATP are needed to transport one molecule of drug Lessons learned from mdr1a/mdr1b knockout mice (Berns, Schinkel, Borst) Mice are fully viable and fertile under controlled lab conditions Mice are very sensitive to toxic xenobiotics, especially those which are neurotoxic Pharmacokinetics of many different P-gp substrates altered: Vinca alkaloids, digoxin, fexofenadine, ivermectin--increased GI absorption, decreased kidney and liver excretion Polymorphisms in the MDR1 gene 5 common coding polymorphisms (Asn21Asp, Phe103Leu, Ser400Asn, Ala892Ser, Ala998Thr) have no demonstrable effect on drug transport functio