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Principal Investigator: THOMAS MELENDY
Organization: STATE UNIVERSITY OF NEW YORK AT BUFFALO
Fiscal Year: 2023
Award: $199,375
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract:
Effective antivirals are sorely needed against SARS CoV-2 (CoV2), and coronaviruses (CoV) in
general. Screening systems for drugs targeting CoV2 enzymes usually involve screening against
purified enzymes. This can be technically challenging, and the vast majority of inhibitors identified
ultimately lack utility as antivirals because: they may not be transported into and retained in cells,
altered within cells such that they no longer inhibit the target effectively, or exhibit off-target effects that
preclude their use as therapeutics. Using predicted drugs directly in viral infection assays can exhibit
off-target effects against the host cell or other viral proteins and can be difficult to distinguish within the
context of the viral infection (the inhibition of CoV2 infection by hydroxychloroquine is one such
example). Ideally screens for viral enzymes isolate the enzyme from the viral infection, but evaluate
function within human cells. We have constructed vectors for expression of four wild-type CoV2
proteins, and unique biosensors, to evaluate function of two CoV2 enzymatic functions, the CoV Main
Protease and the CoV RNA-dependent RNA polymerase complex. We are completing assay validation
and beginning to use the assays to evaluate the few known inhibitors of these enzymes. The protease
assay will be converted to a stable cell line for large scale screening programs. The transfection assays
will be evaluated using both established and experimental 3CL and RdRpC inhibitors, and 128 FDA-
approved drugs that we have predicted target these enzymes, to validate the usefulness of these
screening assays. Drugs that show inhibition of activity will be quantified using dose-response analysis
in the cell-based assays, and subsequently analyzed for inhibition of CoV2 infection of cultured human
lung cells across a similar dose-response range. Following validation, vectors and/or stable cells lines
for the cell-based drug screening systems will be made readily available to other researchers, such as
NIH’s NCATS, who are currently seeking CoV antiviral agents.
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