Structure and inhibition of the SARS-CoV-2 main protease reveals strategy for developing dual inhibitors against M<sup>pro</sup> and cathepsin L

Michael Sacco

Chunlong

Panagiotis Lagarias

Ang Gao

Yanmei Hu+8 authors

^{Department of Molecular Medicine, Morsani College of Medicine, University of South Florida, Tampa, FL, 33612, United States}

^{Department of Pharmacology and Toxicology, College of Pharmacy, The University of Arizona, Tucson, AZ, 85721, United States}

^{Department of Pharmaceutical Chemistry, Faculty of Pharmacy, National and Kapodistrian University of Athens, Athens, 15771, Greece}

^{Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, 85721, United States}

^{Department of Microbiology, Immunology and Molecular Genetics, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229, United States}

^{Institute for Antiviral Research, Utah State University, Logan, UT, 84322, United States}

^{Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, UT, 84322, United States}

Contributed by

AUTHOR CONTRIBUTIONS

J. W. and Y. C. conceived and designed the study; C. M. expressed the M^{and PL}^{; C.M. performed the IC}₅₀ determination, thermal shift-binding assay, and enzymatic kinetic studies; M. S. carried out M^{crystallization and structure determination with the assistance of X. Z, and analyzed the data with Y. C.; P. L. performed the molecular dynamics simulations under the guidance of A. K.; A. G. and N. K. synthesized the compounds for co-crystallization; Y. H. performed the cytotoxicity assay; X. M. and P. D. performed the SARS-CoV-2 immunofluorescence assay and plaque assay under the guidance of Y. X. B. H. and B. T. performed the initial antiviral assay with SARS-CoV-2; J .A. T. performed the native mass spectrometry experiments with the guidance from M. T. M.; J. W. and Y. C. secured funding and supervised the study; J. W., Y.C., and M.S. wrote the manuscript with the input from others.}

^{Corresponding authors: Jun Wang, Tel: 520-626-1366, Fax: 520-626-0749,}ude.anozira.ycamrahp@gnawnuj, Yu Chen, Tel: 813-974-7809, ude.fsu@1nehcy

It is made available under a CC-BY-NC 4.0 International license.

Footnotes

DATA AVAILABILITY. The drug-bound complex structures for SARS-CoV-2 M^{have been deposited in the Protein Data Bank with accession numbers of 6XA4 (SARS-CoV-2 HM}^{+ Calpain inhibitor II), 6XFN (SARS-CoV-2 HM}^{+ Calpain inhibitor XII), 6XBG (SARS-CoV-2 HM}⁺UAWJ246), 6XBH (SARS-CoV-2 HM⁺UAWJ247), and 6XBI (SARS-CoV-2 HM⁺UAWJ248).

ADDITIONAL INFORMATION

Supplementary information accompanies this paper at

Competing interests

The authors declare no competing interests.

Footnotes

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