Biological Evaluation of Molecules of the azaBINOL Class as Antiviral Agents: Specific Inhibition of HIV-1 RNase H Activity by 7-Isopropoxy-8-(naphth-1-yl)quinoline

Ross D. Overacker; Somdev Banerjee; George F. Neuhaus; Selena Milicevic Sephton; Alexander Herrmann; James A. Strother; Ruth Brack-Werner; Paul R. Blakemore; Sandra Loesgen

To show that B#24 inhibits RNase H activity by binding the HIV-1 reverse transcriptase 251 enzyme, we explored direct binding to immobilized HIV-1 RT using bio-layer interferometry 252 (BLI). BLI allows for the real time measurement of binding affinities between ligands and 253 analytes of varying size using single-use, fiber optical sensors. BLI measures association (kon) 254 and dissociation (koff) rates directly from full spectrum wavelength shifts associated with 255 interference pattern changes derived from binding events at a sensors tip to determine binding 256 affinities (KD). 54 Recombinant-wild type HIV-1 p66/p51 RT (NIH Aids Reagents) was 257 immobilized via amine coupling onto BLI biosensors. B#24, selected other azaBINOLs, and 258 control compounds (rilpivirine positive control, raltegravir negative control) were prepared in 1x 259 kinetics buffer at multiple concentrations. Each compound was tested at multiple concentrations, 260 All rights reserved. No reuse allowed without permission.