Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6.

Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Research Abstract Details 

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Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Abstract Text:

Zhongliu-Liu Wu,Pramod Aryal,Olivier Lozach,Laurent Meijer,F Peter Guengerich,

Glycogen synthase kinase-3 (GSK-3) is a potential drug target for a number of human diseases. Some indigoids have been found to be potent inhibitors of GSK-3, and individual compounds with better activity, specificity, and solubility are desired. In this work, a new disubstituted indigoid generation system was developed with a tryptophanase-deficient Escherichia coli strain as a host to express the human cytochrome P450 2A6 mutant L240C/N297Q, which catalyzes the oxidation of indole to isatin and indoxyl, which in turn react to generate indigoids. Forty-five substituted 1H-indoles from commercial sources were used as substrates in the system, and indigoid mixtures were tested as potential inhibitors of GSK-3. After preliminary screening, cell extracts with high inhibitory activity towards GSK-3alpha/beta were fractionated, and the IC50 values of twelve individual indigoids were measured for GSK-3alpha/beta as well as the protein kinases CDK1/cyclinB and CDK5/p25. Several indigoids, including an indigo, showed stronger inhibition than found in previous work. The most potent towards GSK-3alpha/beta, dimethyl indirubin 5,5'-dicarboxylate (IC50 of 51 nM), was modified by chemical reactions. One product, indirubin 5,5'-dicarboxylic acid 5-methyl ester, inhibited GSK-3alpha/beta with an IC50 of 14 nM and selectivity nearly 40-fold over CDK1 and CDK5. Indirubin-5-5'-dicarbonitrile was also modified to the corresponding 3'-oxime, which had low specificity but showed very high inhibition of all three kinases with IC50 values of 5, 13, and 10 nM towards GSK-3alpha/beta, CDK1, and CDK5, respectively. Thus, this system has the potential to generate new indigoids with therapeutic potential.

Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Publishing Authors By Initials

ZL Wu,P Aryal,O Lozach,L Meijer,FP Guengerich,

For similar proteins: recombinant proteins research abstracts see: proteins: recombinant proteins research

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Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Chemistry & biodiversity

VOLUME: 2

Page Numbers: 51-65

Journal Abbreviation: Chem. Biodivers.

ISSN: 1612-1880

DAY: 3

MONTH: Jan

YEAR: 2005

Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101197449

Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Keywords Mesh Terms:

KEYWORDS: Recombinant Proteins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6. Information

Substance Name: Glycogen Synthase Kinase 3

Registry Number: EC 2.7.1.37

Grant and Affiliation Information for Biosynthesis of new indigoid inhibitors of protein kinases using recombinant cytochrome P450 2A6.

AFFILIATION: Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, 638 Robinson Research Building, 23rd and Pierce Avenues, Nashville, Tennessee 37232-0146, USA.

Country: Switzerland

AGENCY: United States NCI

GRANT: R01 CA90426

ACRONYM: CA

MEDLINETA: Chem Biodivers

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