摘要 |
<p>The present invention provides a novel method for the identification of inhibitors of HCV NS5B polymerase that uses an HCV NS5B RNA-dependent RNA polymerase having a higher Km than the full length native NS5B polymerase to ensure identification of inhibitors of HCV polymerase primer-template binding, including moderately inhibitory compounds. The method includes providing a decreased-affinity HCV NS5B polymerase, an appropriate primer-template, and a plurality of appropriate ribonucleotide triphosphates, incubating the HCV NS5B polymerase with the primer-template in the presence and absence of a potential inhibitor, measuring the presence of any polymerase products formed in the presence and absence of the potential inhibitor; and comparing the amount of the polymerase products formed in the presence and absence of the potential inhibitor; wherein a decrease in the amount of the polymerase products formed in the presence of the potential inhibitor compared to the amount of polymerase products formed in the absence of the potential inhibitor is indicative of a potential primer-template binding inhibitor of HCV NS5B RNA dependent RNA polymerase.</p> |