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Generation of new protein kinase inhibitors utilizing cytochrome p450 mutant enzymes for indigoid synthesis.
- Source :
-
Journal of medicinal chemistry [J Med Chem] 2004 Jun 03; Vol. 47 (12), pp. 3236-41. - Publication Year :
- 2004
-
Abstract
- Indigoids, a class of bis-indoles, represent a promising protein kinase inhibitor scaffold. Oxidation of indole by cytochrome P450 (P450) has been shown to generate species (indoxyl, isatin) that couple to yield indigo and indirubin. Escherichia coli-expressed human P450 2A6 mutants isolated from a randomized library were incubated with 27 substituted indole derivatives. Extracts of the cultures were screened for inhibition of human cyclin-dependent kinases (CDK)-1 and -5 and glycogen synthase kinase-3 (GSK3). The extracts from cultures incubated with 5-methoxyindole were the most inhibitory. High-performance liquid chromatography (HPLC) separation yielded a mixture of seven colored indigoids. These indigoids included indigo, indirubin, the di(5-methoxy) derivatives of indigo and indirubin, and both of the possible mono 5-methoxy derivatives of indirubin, which were all identified by visible, mass, and NMR spectra. Cultures with 5-methylindole added to the media also yielded inhibitory material, and 5- and 5'-methylindirubin were characterized. The most inhibitory of these indigoids were the monosubstituted indirubins and 5,5'-dimethoxyindirubin, which was > or =10x more active than indirubin. Thus, the overall approach involves the use of a library of randomized enzyme mutants to activate component moieties of a desired set of larger molecules, thus yielding a library of drug candidates that can be screened and characterized. The general strategy may have additional applications.
- Subjects :
- Aryl Hydrocarbon Hydroxylases genetics
Aryl Hydrocarbon Hydroxylases isolation & purification
CDC2 Protein Kinase antagonists & inhibitors
Chromatography, High Pressure Liquid
Cyclin-Dependent Kinase 5
Cyclin-Dependent Kinases antagonists & inhibitors
Cytochrome P-450 CYP2A6
Escherichia coli metabolism
Glycogen Synthase Kinase 3 antagonists & inhibitors
Humans
Indoles chemistry
Indoles isolation & purification
Mixed Function Oxygenases genetics
Mixed Function Oxygenases isolation & purification
Mutation
Aryl Hydrocarbon Hydroxylases metabolism
Indoles metabolism
Mixed Function Oxygenases metabolism
Protein Kinase Inhibitors
Subjects
Details
- Language :
- English
- ISSN :
- 0022-2623
- Volume :
- 47
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Journal of medicinal chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 15163202
- Full Text :
- https://doi.org/10.1021/jm030561b