| dc.description.abstract | SARS-CoV-2 has caused a global pandemic since emerged from Wuhan, China. The Main protease of this virus is an important target for drug discovery. To test the real inhibition effect of the developed small molecule inhibitors to Mpro, live-virus based assay is normally used. However, this can be problematic because a lot of other proteases that play important roles in the life cycle of this virus share a similar catalytic mechanism with Mpro, thus would be inhibited by these inhibitors as well. The live-virus based assay is also tedious, dangerous, and requires BSL-3 level laboratory which is not equipped by many institutions. We developed a cellular assay based on the cellular toxicity of Mpro, utilizing a Mpro-eGFP construct that can be directly quantified by flow cytometry to test the inhibition effect of inhibitors developed.
Combined with the cellular assay we developed, we systematically studied the inhibition effect for Mpro of a series of dipeptidyl inhibitors we developed using enzymatic inhibition assays, X-ray crystallization, live-virus based assays and cytotoxicity and in cell stability. The S2 binding pocket of Mpro was found to have a potential to accommodate larger and complicate binding groups and two compounds, MPI60 and MPI61 was selected as two compounds with the most significant potential for clinical study.
Chimeric antigen receptor (CAR) T cell therapy has shown its enormous ability in cancer treatment, while the uncontrollable T cell activation arise in potential of serious side effects. Many efforts have been done to control the activation of CAR-T cells but with obvious drawbacks. We propose a chemogenetic recurring switch on the basis of the third generation CAR design, using HCV-NS3 as the switch and ASV as the regulator. Compared to the standard CAR19 product, this switchable CAR-T design displayed excellent tumor killing effect both in vitro and in vivo. | |
