Efficient screening of cytochrome P450 BM3 mutants for their metabolic activity and diversity toward a wide set of drug-like molecules in chemical space.

Details

Serval ID
serval:BIB_7910DCEABC05
Type
Article: article from journal or magazin.
Collection
Publications
Title
Efficient screening of cytochrome P450 BM3 mutants for their metabolic activity and diversity toward a wide set of drug-like molecules in chemical space.
Journal
Drug metabolism and disposition
Author(s)
Reinen J., van Leeuwen J.S., Li Y., Sun L., Grootenhuis P.D., Decker C.J., Saunders J., Vermeulen N.P., Commandeur J.N.
ISSN
1521-009X (Electronic)
ISSN-L
0090-9556
Publication state
Published
Issued date
09/2011
Peer-reviewed
Oui
Volume
39
Number
9
Pages
1568-1576
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
In the present study, the diversity of a library of drug-metabolizing bacterial cytochrome P450 (P450) BM3 mutants was evaluated by a liquid chromatography-mass spectrometry (LC-MS)-based screening method. A strategy was designed to identify a minimal set of BM3 mutants that displays differences in regio- and stereoselectivities and is suitable to metabolize a large fraction of drug chemistry space. We first screened the activities of six structurally diverse BM3 mutants toward a library of 43 marketed drugs (encompassing a wide range of human P450 phenotypes, cLogP values, charges, and molecular weights) using a rapid LC-MS method with an automated method development and data-processing system. Significant differences in metabolic activity were found for the mutants tested and based on this drug library screen; nine structurally diverse probe drugs were selected that were subsequently used to study the metabolism of a library of 14 BM3 mutants in more detail. Using this alternative screening strategy, we were able to select a minimal set of BM3 mutants with high metabolic activities and diversity with respect to substrate specificity and regiospecificity that could produce both human relevant and BM3 unique drug metabolites. This panel of four mutants (M02, MT35, MT38, and MT43) was capable of producing P450-mediated metabolites for 41 of the 43 drugs tested while metabolizing 77% of the drugs by more than 20%. We observed this as the first step in our approach to use of bacterial P450 enzymes as general reagents for lead diversification in the drug development process and the biosynthesis of drug(-like) metabolites.
Keywords
Bacterial Proteins/chemistry, Bacterial Proteins/genetics, Bacterial Proteins/metabolism, Chromatography, Liquid/methods, Cytochrome P-450 Enzyme System/chemistry, Cytochrome P-450 Enzyme System/genetics, Cytochrome P-450 Enzyme System/metabolism, Gene Library, Humans, Inactivation, Metabolic, Mass Spectrometry/methods, Microsomes, Liver/chemistry, Microsomes, Liver/enzymology, Microsomes, Liver/metabolism, Mutagenesis, Site-Directed, Pharmaceutical Preparations/metabolism, Substrate Specificity
Pubmed
Web of science
Create date
22/01/2019 17:13
Last modification date
21/08/2019 6:35
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