P450 BM3 Catalysis Mechanism and Application in Drug D：细胞色素P450 BM3催化机理及其在药物研发中的应用.ppt

QM/MM Study of Cytochrome P450 BM3 Catalysis Mechanism and Application in Drug Design P450: Super family of monooxygenases Function: metabolism of carbon-source organic molecules: hormones, vitamin D etc. detoxification of xenobiotic compounds synthesis of biologically active compounds Why interesting in drug design The eve of ADME in High-Throughput Screening Absorption by the intestine Diffusion to the organism Metabolism by the liver Excretion by the kidney Toxicology One-half of potential drugs fail because of ADME/Tox issues Of the estimated $600 million cost of bringing a new drug to market, more than $400 million of that is wasted pursuing leads that turn out to be dogs. P450 (CYP) enzymes--a class of enzymes responsible for the metabolism of more than 50% of all known drugs Enzyme P450 BM3 The function of P450 enzyme in drug metabolism is responsible for the failure of a good inhibitor to become a useful drug P450 BM3: the closet bacterial analog to the mammalian enzyme, soluble, three domains (BMP heme domain, a FAD- and FMN-containing NADPH-cytochrome P450 reductase) Putative conformational change Electron transfer: NADPH-FAD-FMN -BMP (heme) Conformational change facilitated by the “hinge” moving between a “closed” conformation and an “open” conformation FMN-heme distance:18?-8? Dutton’s line: electron transfer Productive electron transfer: within 14–15 ? X-ray structure and product Computational modeling Solid State NMR Experiment 2D solid-state NMR 15N13CO SPECIFIC CP spectrum of 13COLeu; 15N-Gly, 15N-Phe labeled cytochrome P450 BM-3 bound with NPG at 0 (red) and -30 °C (blue). The L86-F87 pair exhibits a pronounced shift as a function of temperature. The L86-F87 pair is in the binding pocket, and in fact F87 plays a “gatekeeper” role in that its bulky side chain must be rotated in order to allow for substrate binding. QM/MM Combined Quantum mechanics/ Molecular mechanics (QM/MM) is a hybrid