Identification of Small-molecule Scaffolds for P450 Inhibitors

Overview

Journal Curr Protoc Microbiol

Specialty Microbiology

Date 2010 Feb 5

PMID 20131225

Citations 9

Authors

Jens P von Kries

Thulasi Warrier

Larissa M Podust

Affiliations

Soon will be listed here.

Abstract

Mycobacterium tuberculosis cytochrome P450 enzymes (CYP) attract ongoing interest for their pharmacological development potential, driving direct screening efforts against potential CYP targets with the ultimate goal of developing potent CYP-specific inhibitors and/or molecular probes to address M. tuberculosis biology. The property of CYP enzymes to shift the ferric heme Fe Soret band in response to ligand binding provides the basis for an experimental platform for high-throughput screening (HTS) of compound libraries to select chemotypes with high binding affinities to the target. Promising compounds can be evaluated in in vitro assays or in vivo disease models and further characterized by x-ray crystallography, leading to optimization strategies to assist drug design. Protocols are provided for compound library screening, analysis of inhibitory potential, and co-crystallization with the target CYP, as well as expression and purification of soluble CYP enzymes.

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References

Ahmad Z, Sharma S, Khuller G . In vitro and ex vivo antimycobacterial potential of azole drugs against Mycobacterium tuberculosis H37Rv. FEMS Microbiol Lett. 2005; 251(1):19-22. DOI: 10.1016/j.femsle.2005.07.022. View

Sheehan D, Hitchcock C, Sibley C . Current and emerging azole antifungal agents. Clin Microbiol Rev. 1999; 12(1):40-79. PMC: 88906. DOI: 10.1128/CMR.12.1.40. View

Schenkman J, Remmer H, Estabrook R . Spectral studies of drug interaction with hepatic microsomal cytochrome. Mol Pharmacol. 1967; 3(2):113-23. View

Chen C, Doyle P, Yermalitskaya L, Mackey Z, Ang K, McKerrow J . Trypanosoma cruzi CYP51 inhibitor derived from a Mycobacterium tuberculosis screen hit. PLoS Negl Trop Dis. 2009; 3(2):e372. PMC: 2629123. DOI: 10.1371/journal.pntd.0000372. View

Dunford A, McLean K, Sabri M, Seward H, Heyes D, Scrutton N . Rapid P450 heme iron reduction by laser photoexcitation of Mycobacterium tuberculosis CYP121 and CYP51B1. Analysis of CO complexation reactions and reversibility of the P450/P420 equilibrium. J Biol Chem. 2007; 282(34):24816-24. DOI: 10.1074/jbc.M702958200. View

Hendrickson W, Horton J, Murthy H, Pahler A, Smith J . Multiwavelength anomalous diffraction as a direct phasing vehicle in macromolecular crystallography. Basic Life Sci. 1989; 51:317-24. DOI: 10.1007/978-1-4684-8041-2_28. View

Nwaka S, Hudson A . Innovative lead discovery strategies for tropical diseases. Nat Rev Drug Discov. 2006; 5(11):941-55. DOI: 10.1038/nrd2144. View

Cali J, Ma D, Sobol M, Simpson D, Frackman S, Good T . Luminogenic cytochrome P450 assays. Expert Opin Drug Metab Toxicol. 2006; 2(4):629-45. DOI: 10.1517/17425255.2.4.629. View

Pieters J . Mycobacterium tuberculosis and the macrophage: maintaining a balance. Cell Host Microbe. 2008; 3(6):399-407. DOI: 10.1016/j.chom.2008.05.006. View

10.

Fai P, Grant A . A rapid resazurin bioassay for assessing the toxicity of fungicides. Chemosphere. 2009; 74(9):1165-70. DOI: 10.1016/j.chemosphere.2008.11.078. View

11.

Banfi E, Scialino G, Zampieri D, Mamolo M, Vio L, Ferrone M . Antifungal and antimycobacterial activity of new imidazole and triazole derivatives. A combined experimental and computational approach. J Antimicrob Chemother. 2006; 58(1):76-84. DOI: 10.1093/jac/dkl182. View

12.

Cole S, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D . Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998; 393(6685):537-44. DOI: 10.1038/31159. View

13.

Ahmad Z, Sharma S, Khuller G . Azole antifungals as novel chemotherapeutic agents against murine tuberculosis. FEMS Microbiol Lett. 2006; 261(2):181-6. DOI: 10.1111/j.1574-6968.2006.00350.x. View

14.

Yajko D, Madej J, Lancaster M, Sanders C, Cawthon V, Gee B . Colorimetric method for determining MICs of antimicrobial agents for Mycobacterium tuberculosis. J Clin Microbiol. 1995; 33(9):2324-7. PMC: 228403. DOI: 10.1128/jcm.33.9.2324-2327.1995. View

15.

Holton J . A beginner's guide to radiation damage. J Synchrotron Radiat. 2009; 16(Pt 2):133-42. PMC: 2651760. DOI: 10.1107/S0909049509004361. View

16.

Flynn J . Immunology of tuberculosis and implications in vaccine development. Tuberculosis (Edinb). 2003; 84(1-2):93-101. DOI: 10.1016/j.tube.2003.08.010. View

17.

Podust L, Yermalitskaya L, Lepesheva G, Podust V, Dalmasso E, Waterman M . Estriol bound and ligand-free structures of sterol 14alpha-demethylase. Structure. 2004; 12(11):1937-45. DOI: 10.1016/j.str.2004.08.009. View

18.

Podust L, von Kries J, Eddine A, Kim Y, Yermalitskaya L, Kuehne R . Small-molecule scaffolds for CYP51 inhibitors identified by high-throughput screening and defined by X-ray crystallography. Antimicrob Agents Chemother. 2007; 51(11):3915-23. PMC: 2151439. DOI: 10.1128/AAC.00311-07. View

19.

Byrne S, Denkin S, Gu P, Nuermberger E, Zhang Y . Activity of ketoconazole against Mycobacterium tuberculosis in vitro and in the mouse model. J Med Microbiol. 2007; 56(Pt 8):1047-1051. DOI: 10.1099/jmm.0.47058-0. View

20.

Saunders B, Britton W . Life and death in the granuloma: immunopathology of tuberculosis. Immunol Cell Biol. 2007; 85(2):103-11. DOI: 10.1038/sj.icb.7100027. View