Computer aided design of selective cytochrome P450 inhibitors

I have generated a virtual library of imidazole based compounds and used an altered form of the DOCK algorithm, CovDOCK, to screen these compounds against heme-containing proteins. I was able to generate high affinity ligands for P450cam. An L244A mutant of P450cam was generated and used to test our ability to generate selective ligands. Using our methods I was able to generate ligands which only inhibit the mutant enzyme.; The library was expanded with more substituted imidazoles and screened against a viable drug target, Mycobacterium tuberculosis CYP51. After one round of compound development I was able to find micromolar inhibitors. I also demonstrated that the methods and library are easily applicable to new protein systems.; To explore differences of the imidazole compounds in binding and inhibition, I carried out stopped flow kinetics experiments and further UV spectroscopic studies. I find that mono- and disubstituted imidazoles bind distinctly to wild-type P450cam. The natural substrate also affects both classes of ligands uniquely.; To explore the mitigated affinity for imidazole of the L244A mutant, I have done crystallography studies. I report the structures of the substrate-free mutant, and imidazole co-crystallized wild-type and mutant proteins. I present a possible explanation for the mitigated imidazole affinity based on structural factors observed when juxtaposing the wild-type and mutant structures.