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Inhibition of Flaviviridae Replication Complex: Assays to Investigate the Modulating Effect of Potential Compounds

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Approximately up to 200 million new cases of infections caused by viruses of the Flaviviridae family, like hepatitis C virus (HCV), West-Nile virus (WNV), dengue virus (DENV) and Japanese encephalitis virus (JEV) are registered annually.

Up to date, there is no effective antiviral therapy directed against Flaviviridae viruses. For the replication of all viruses the intact function of nonstructural proteins is necessary. The blockade of their activities leads to inhibition of the virus propagation. Thus, the nucleotide triphosphatase (NTPase) as well as helicase activities of NS3 and the RNA-dependent RNA polymerase (RdRp) activity of NS5 (or NS5B) appear to be exceptionally attractive targets for antiviral compounds. In this context, a systematic screening of chemical libraries and computational analysis are necessary which will lead to the development of effective inhibitors of the Flaviviridae replication complex.

In this review the most promissing methods based on different techniques to quantify the enzymatic activities, like radioactivity, fluorescence, colorimetry as well as a coupled second enzymatic reaction are summarized and discussed. These methods serve as a starting point to establish a high throughput system (HTS) that permits more efficient screenings.

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Keywords: Flaviviridae; NTPase/helicase; RNA-dependent RNA polymerase; in vitro assays

Document Type: Research Article

Publication date: 2009-12-01

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  • Current Enzyme Inhibition aims to publish all the latest and outstanding developments in enzyme inhibition studies with regards to the mechanisms of inhibitory processes of enzymes, recognition of active sites, and the discovery of agonists and antagonists, leading to the design and development of new drugs of significant therapeutic value. Each issue contains a series of timely, in-depth reviews written by leaders in the field, covering a range of enzymes that can be exploited for drug development. Current Enzyme Inhibition is an essential journal for every pharmaceutical and medicinal chemist who wishes to have up-to-date knowledge about each and every development in the study of enzyme inhibition.
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