Identification of small molecule inhibitors of TAP protein

National Center For Advancing Translational Sciences

Investigators

Abstract

In our previous pilot screening and validation of NPC library, we found that all hits are false positive, which is either the inhibitors of NanoBiT luciferase or toxic compounds to the cells. To triage these false positives, we developed and optimized two additional assays for TAP1 project: 1. cell-based counter assay, which LgBiT was targeted in the cytosol instead of endoplasmic reticulum. 2. Biochemical-based counter assay, which we can differentiate whether the hits are inhibitors of NanoBiT luciferase. We screened two more libraries (NPACT and Syntravon Mimime) in one concentration (10uM final concentration) in the primary assay we developed. 508 hits were cherrypicked using the criteria (>50% activity). After confirmation assay and two counter assays in 11 dose, 21 hits were confirmed and sent to our collaborator for validation. 11 hits were further confirmed in our collaborator’s lab. Our collaborator is particularly interested in one hit and is validating it in multiple assays they developed in their lab.