Direct binding assay for identifying inhibitors of HCV polymerase

摘要

A method for identifying compounds binding to HCV polymerase comprising the steps of: contacting said HCV polymerase or an analog thereof with a probe formula I: wherein A is O, S, N, NR¹, or CR¹, wherein R¹ is defined herein; ----- represents either a single or a double bond; R² is selected from: H, halogen, R²¹, OR²¹, SR²¹, COOR²¹, SO₂N(R²²)₂, N(R²²)₂, CON(R²²)₂, NR²²C(O)R²² or NR²²C(O)NR²² wherein R²¹ and each R²² is defined herein; B is NR³ or CR³, wherein R³ is defined herein; with the proviso that, when A is not N, then one of A or B is either CR¹ or CR³, K is N or CR⁴, wherein R⁴ is defined herein; L is N or CR⁵, wherein R⁵ has the same definition as R⁴ defined above; M is N or CR⁷, wherein R⁷ has the same definition as R⁴ defined above; R⁵ is C(Y¹)Z wherein Y¹ is O or S; and Z is N(R^6a)R⁶ or OR⁶, wherein R^6a is H or alkyl or NR⁶¹R⁶² wherein R⁶¹ and R⁶² are defined herein; and R⁶ is H, alkyl, cycloalkyl, alkenyl, Het, alkyl-aryl, alkyl-Het; or R⁶ is wherein R⁷ and R⁸ and Q are as defined herein; Y² is O or S; R⁹ is H, (C_1-6 alkyl), (C_3-7)cycloalkyl or (C_1-6)alkyl-(C_3-7)cycloalkyl, aryl, Het, (C_1-6)alkyl-aryl or (C_1-6)alkyl-Het, all of which optionally substituted with R⁹⁰; or R⁹ is covalently bonded to either of R⁷ or R⁸ to form a 5- or 6-membered heterocycle; or a salt thereof; where the probe comprises a detectable label attached to any suitable position, whereby said probe binds to an HCV polymerase or an analog thereof and is capable of being displaced by an inhibitor thereof.