New Non-nucleoside Inhibitors of Hepatitis C Virus RNA-Dependent RNA Polymerase

A. V. Ivanov^1*, M. V. Kozlov², A. O. Kuzyakin¹, D. A. Kostyuk¹, V. L. Tunitskaya¹, and S. N. Kochetkov¹

¹Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow 119991, Russia; fax: (7-095) 135-1405; E-mail: aivanov@yandex.ru

²Institute of Limnology, Siberian Division of the Russian Academy of Sciences, ul. Ulan-Batorskaya 3, Irkutsk 664033, Russia; fax: (3-952) 42-5405

^* To whom correspondence should be addressed.

Received December 19, 2003; Revision received February 10, 2004

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Recombinant RNA-dependent RNA polymerase of hepatitis C virus was purified using a bacterial expression system (Escherichia coli). The system for enzyme activity detection was optimized. The maximum activity was achieved when the reaction was carried out at 30°C in the presence of 3 mM Mg²⁺ or 0.75 mM Mn²⁺. Among alpha- and beta-pyrogallaldehydes, effective inhibitors were found. It was shown that they acted at the primer elongation stage, and their binding to the protein is reversible.

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KEY WORDS: hepatitis C virus, RNA-dependent RNA polymerase, non-nucleoside inhibitors

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