Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19

Britton Boras; Rhys M. Jones; Brandon J. Anson; Dan Arenson; Lisa Aschenbrenner; Malina A. Bakowski; Nathan Beutler; Joseph Binder; Emily Chen; Heather Eng; Holly Hammond; Jennifer Hammond; Robert E. Haupt; Robert Hoffman; Eugene P. Kadar; Rob Kania; Emi Kimoto; Melanie G. Kirkpatrick; Lorraine Lanyon; Emma K. Lendy; Jonathan R. Lillis; James Logue; Suman A. Luthra; Chunlong Ma; Stephen W. Mason; Marisa E. McGrath; Stephen Noell; R. Scott Obach; Matthew N. O’ Brien; Rebecca O’Connor; Kevin Ogilvie; Dafydd Owen; Martin Pettersson; Matthew R. Reese; Thomas F. Rogers; Romel Rosales; Michelle I. Rossulek; Jean G. Sathish; Norimitsu Shirai; Claire Steppan; Martyn Ticehurst; Lawrence W. Updyke; Stuart Weston; Yuao Zhu; Kris M. White; Adolfo García-Sastre; Jun Wang; Arnab K. Chatterjee; Andrew D. Mesecar; Matthew B. Frieman; Annaliesa S. Anderson; Charlotte Allerton

doi:10.1038/s41467-021-26239-2

Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19

Britton Boras, Rhys M. Jones, Brandon J. Anson, Dan Arenson, Lisa Aschenbrenner, Malina A. Bakowski, Nathan Beutler, Joseph Binder, Emily Chen, Heather Eng, Holly Hammond, Jennifer Hammond, Robert E. Haupt, Robert Hoffman, Eugene P. Kadar, Rob Kania, Emi Kimoto, Melanie G. Kirkpatrick, Lorraine Lanyon, Emma K. LendyJonathan R. Lillis, James Logue, Suman A. Luthra, Chunlong Ma, Stephen W. Mason, Marisa E. McGrath, Stephen Noell, R. Scott Obach, Matthew N. O’ Brien, Rebecca O’Connor, Kevin Ogilvie, Dafydd Owen, Martin Pettersson, Matthew R. Reese, Thomas F. Rogers, Romel Rosales, Michelle I. Rossulek, Jean G. Sathish, Norimitsu Shirai, Claire Steppan, Martyn Ticehurst, Lawrence W. Updyke, Stuart Weston, Yuao Zhu, Kris M. White, Adolfo García-Sastre, Jun Wang, Arnab K. Chatterjee, Andrew D. Mesecar, Matthew B. Frieman, Annaliesa S. Anderson, Charlotte Allerton

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

COVID-19 caused by the SARS-CoV-2 virus has become a global pandemic. 3CL protease is a virally encoded protein that is essential across a broad spectrum of coronaviruses with no close human analogs. PF-00835231, a 3CL protease inhibitor, has exhibited potent in vitro antiviral activity against SARS-CoV-2 as a single agent. Here we report, the design and characterization of a phosphate prodrug PF-07304814 to enable the delivery and projected sustained systemic exposure in human of PF-00835231 to inhibit coronavirus family 3CL protease activity with selectivity over human host protease targets. Furthermore, we show that PF-00835231 has additive/synergistic activity in combination with remdesivir. We present the ADME, safety, in vitro, and in vivo antiviral activity data that supports the clinical evaluation of PF-07304814 as a potential COVID-19 treatment.

Original language	English (US)
Article number	6055
Journal	Nature communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-26239-2
State	Published - Dec 2021

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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10.1038/s41467-021-26239-2

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Boras, B., Jones, R. M., Anson, B. J., Arenson, D., Aschenbrenner, L., Bakowski, M. A., Beutler, N., Binder, J., Chen, E., Eng, H., Hammond, H., Hammond, J., Haupt, R. E., Hoffman, R., Kadar, E. P., Kania, R., Kimoto, E., Kirkpatrick, M. G., Lanyon, L., ... Allerton, C. (2021). Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19. Nature communications, 12(1), [6055]. https://doi.org/10.1038/s41467-021-26239-2

Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19. / Boras, Britton; Jones, Rhys M.; Anson, Brandon J.; Arenson, Dan; Aschenbrenner, Lisa; Bakowski, Malina A.; Beutler, Nathan; Binder, Joseph; Chen, Emily; Eng, Heather; Hammond, Holly; Hammond, Jennifer; Haupt, Robert E.; Hoffman, Robert; Kadar, Eugene P.; Kania, Rob; Kimoto, Emi; Kirkpatrick, Melanie G.; Lanyon, Lorraine; Lendy, Emma K.; Lillis, Jonathan R.; Logue, James; Luthra, Suman A.; Ma, Chunlong; Mason, Stephen W.; McGrath, Marisa E.; Noell, Stephen; Obach, R. Scott; O’ Brien, Matthew N.; O’Connor, Rebecca; Ogilvie, Kevin; Owen, Dafydd; Pettersson, Martin; Reese, Matthew R.; Rogers, Thomas F.; Rosales, Romel; Rossulek, Michelle I.; Sathish, Jean G.; Shirai, Norimitsu; Steppan, Claire; Ticehurst, Martyn; Updyke, Lawrence W.; Weston, Stuart; Zhu, Yuao; White, Kris M.; García-Sastre, Adolfo; Wang, Jun; Chatterjee, Arnab K.; Mesecar, Andrew D.; Frieman, Matthew B.; Anderson, Annaliesa S.; Allerton, Charlotte.

In: Nature communications, Vol. 12, No. 1, 6055, 12.2021.

Research output: Contribution to journal › Article › peer-review

Boras, B, Jones, RM, Anson, BJ, Arenson, D, Aschenbrenner, L, Bakowski, MA, Beutler, N, Binder, J, Chen, E, Eng, H, Hammond, H, Hammond, J, Haupt, RE, Hoffman, R, Kadar, EP, Kania, R, Kimoto, E, Kirkpatrick, MG, Lanyon, L, Lendy, EK, Lillis, JR, Logue, J, Luthra, SA, Ma, C, Mason, SW, McGrath, ME, Noell, S, Obach, RS, O’ Brien, MN, O’Connor, R, Ogilvie, K, Owen, D, Pettersson, M, Reese, MR, Rogers, TF, Rosales, R, Rossulek, MI, Sathish, JG, Shirai, N, Steppan, C, Ticehurst, M, Updyke, LW, Weston, S, Zhu, Y, White, KM, García-Sastre, A, Wang, J, Chatterjee, AK, Mesecar, AD, Frieman, MB, Anderson, AS & Allerton, C 2021, 'Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19', Nature communications, vol. 12, no. 1, 6055. https://doi.org/10.1038/s41467-021-26239-2

Boras, Britton ; Jones, Rhys M. ; Anson, Brandon J. ; Arenson, Dan ; Aschenbrenner, Lisa ; Bakowski, Malina A. ; Beutler, Nathan ; Binder, Joseph ; Chen, Emily ; Eng, Heather ; Hammond, Holly ; Hammond, Jennifer ; Haupt, Robert E. ; Hoffman, Robert ; Kadar, Eugene P. ; Kania, Rob ; Kimoto, Emi ; Kirkpatrick, Melanie G. ; Lanyon, Lorraine ; Lendy, Emma K. ; Lillis, Jonathan R. ; Logue, James ; Luthra, Suman A. ; Ma, Chunlong ; Mason, Stephen W. ; McGrath, Marisa E. ; Noell, Stephen ; Obach, R. Scott ; O’ Brien, Matthew N. ; O’Connor, Rebecca ; Ogilvie, Kevin ; Owen, Dafydd ; Pettersson, Martin ; Reese, Matthew R. ; Rogers, Thomas F. ; Rosales, Romel ; Rossulek, Michelle I. ; Sathish, Jean G. ; Shirai, Norimitsu ; Steppan, Claire ; Ticehurst, Martyn ; Updyke, Lawrence W. ; Weston, Stuart ; Zhu, Yuao ; White, Kris M. ; García-Sastre, Adolfo ; Wang, Jun ; Chatterjee, Arnab K. ; Mesecar, Andrew D. ; Frieman, Matthew B. ; Anderson, Annaliesa S. ; Allerton, Charlotte. / Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19. In: Nature communications. 2021 ; Vol. 12, No. 1.

@article{1b45f56d1f0448d8b762c8c36799b6ed,

title = "Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19",

abstract = "COVID-19 caused by the SARS-CoV-2 virus has become a global pandemic. 3CL protease is a virally encoded protein that is essential across a broad spectrum of coronaviruses with no close human analogs. PF-00835231, a 3CL protease inhibitor, has exhibited potent in vitro antiviral activity against SARS-CoV-2 as a single agent. Here we report, the design and characterization of a phosphate prodrug PF-07304814 to enable the delivery and projected sustained systemic exposure in human of PF-00835231 to inhibit coronavirus family 3CL protease activity with selectivity over human host protease targets. Furthermore, we show that PF-00835231 has additive/synergistic activity in combination with remdesivir. We present the ADME, safety, in vitro, and in vivo antiviral activity data that supports the clinical evaluation of PF-07304814 as a potential COVID-19 treatment.",

author = "Britton Boras and Jones, {Rhys M.} and Anson, {Brandon J.} and Dan Arenson and Lisa Aschenbrenner and Bakowski, {Malina A.} and Nathan Beutler and Joseph Binder and Emily Chen and Heather Eng and Holly Hammond and Jennifer Hammond and Haupt, {Robert E.} and Robert Hoffman and Kadar, {Eugene P.} and Rob Kania and Emi Kimoto and Kirkpatrick, {Melanie G.} and Lorraine Lanyon and Lendy, {Emma K.} and Lillis, {Jonathan R.} and James Logue and Luthra, {Suman A.} and Chunlong Ma and Mason, {Stephen W.} and McGrath, {Marisa E.} and Stephen Noell and Obach, {R. Scott} and {O{\textquoteright} Brien}, {Matthew N.} and Rebecca O{\textquoteright}Connor and Kevin Ogilvie and Dafydd Owen and Martin Pettersson and Reese, {Matthew R.} and Rogers, {Thomas F.} and Romel Rosales and Rossulek, {Michelle I.} and Sathish, {Jean G.} and Norimitsu Shirai and Claire Steppan and Martyn Ticehurst and Updyke, {Lawrence W.} and Stuart Weston and Yuao Zhu and White, {Kris M.} and Adolfo Garc{\'i}a-Sastre and Jun Wang and Chatterjee, {Arnab K.} and Mesecar, {Andrew D.} and Frieman, {Matthew B.} and Anderson, {Annaliesa S.} and Charlotte Allerton",

note = "Funding Information: The authors would like to thank Sarah Lazzaro, Sumathy Mathialagan, Sangwoo Ryu, Mark West and Emi Yamaguchi (Pfizer) for the transporter inhibition studies. Angela Doran, Chad Limanni, Amanda Plante and Jocelyn Rosado for their in vivo and PK study support (Pfizer). Marcus Ewing (Pfizer) and Gail Johnson (Pfizer) for preformulation studies. Li Hao (Pfizer) for sequence analysis support. Shinji Yamazaki (Pfizer) for PBPK modeling simulations. Daniel Lettiere, Michael Homiski, Michelle Kenyon, Asser Bas-syouni, Declan Flynn, William Reagan, Victoria Markiewicz and Stephen Jenkinson for overseeing safety studies, and William Reagan, for expert clinical pathology and pathology support for the toxicology studies. Deli Huang for supplying the HeLa-ACE2 stably transfected cell line. R. Albert for support with the BSL3 facility and procedures at the Icahn School of Medicine at Mount Sinai, New York. Devendra Rai (Pfizer) and Charles Tan (Pfizer) for their help with the editorial process and statistics. Andrew Mesecar acknowledges partial support for this project from federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Contract No. HHSN272201700060C. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The study performed in Dr. Jun Wang{\textquoteright}s laboratory is partially supported by NIH grant (AI147325) and the Young Investigator Award grant from the Arizona Biomedical Research Centre (ADHS18-198859). Scripps work was supported by a grant from the Bill & Melinda Gates Foundation #OPP1107194, and the Scripps Family Impact Fund of the Miramar Charitable Foundation (MCF) MBF is supported by NIH grants R21AI158134, R01 AI148166, R21AI153480, HHSN272201400007C and 75N93019C00051. A DARPA subcontract #HR0011-20-2-0040, BARDA contract #ASPR-20-01495, and Bill and Melinda Gates Foundation grants # INV-006099 and INV-016638. This research was also partly funded by CRIP (Center for Research for Influenza Pathogenesis), a NIAID supported Center of Excellence for Influenza Research and Surveillance (CEIRS, contract # HHSN272201400008C), by DARPA grant HR0011-19-2-0020, by supplements to NIAID grant U19AI142733 U19AI135972 and DoD grant W81XWH-20-1-0270, and by the generous support of the JPB Foundation, the Open Philanthropy Project (research grant 2020-215611 (5384)), and anonymous donors to AG-S. Funding Information: A.D.M. and affiliates (B.J.A., E.K.L.) have a sponsored program contract with Pfizer to test compounds for inhibition of coronavirus proteases. J.W. and affiliate (C.M.) have a sponsored research agreement with Pfizer to test compounds for inhibition of coronavirus proteases. The Frieman Laboratory (M.B.F., H.H., R.E.Ha., J.L., M.E.M., S.W.) was funded by Pfizer for the work in this manuscript. The Garc{\'i}a-Sastre Laboratory (AGS, KMW, RR) has received research support from Pfizer, Senhwa Biosciences, Accurius, Avimex, 7Hills Pharma, Pharmamar, Blade therapeutics, Dynavax, Kenall Manufacturing, ImmunityBio and Nanocomposix; and AG-S has consulting agreements for the following companies involving cash and/or stock: Vivaldi Biosciences, Contrafect, 7Hills Pharma, Avimex, Valneva, Accurius, and Esperovax. AG-S is inventor in patent applications on SARS-CoV-2 antivirals owned by Icahn School of Medicine at Mount Sinai. B.B., R.M.J., D.A., L.A., J.B., H.E., J.H., R.Ho., E.P.K., R.K., E.K., L.L., J.R.L., S.A.L., S.W.M., S.N., R.S.O., M.N.O., R.O., K.O., D.O., M.P., M.R.R., M.I.R., J.G.S., N.S., C.S., M.T., L.W.U., Y.Z., A.A., and C.A., at the time of their contributions were employees and may have stock in Pfizer. A.K.C., M.A.B., N.B., E.C., M.G.K., T.F.R. have no competing interest to report. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = dec,

doi = "10.1038/s41467-021-26239-2",

language = "English (US)",

volume = "12",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19

AU - Boras, Britton

AU - Jones, Rhys M.

AU - Anson, Brandon J.

AU - Arenson, Dan

AU - Aschenbrenner, Lisa

AU - Bakowski, Malina A.

AU - Beutler, Nathan

AU - Binder, Joseph

AU - Chen, Emily

AU - Eng, Heather

AU - Hammond, Holly

AU - Hammond, Jennifer

AU - Haupt, Robert E.

AU - Hoffman, Robert

AU - Kadar, Eugene P.

AU - Kania, Rob

AU - Kimoto, Emi

AU - Kirkpatrick, Melanie G.

AU - Lanyon, Lorraine

AU - Lendy, Emma K.

AU - Lillis, Jonathan R.

AU - Logue, James

AU - Luthra, Suman A.

AU - Ma, Chunlong

AU - Mason, Stephen W.

AU - McGrath, Marisa E.

AU - Noell, Stephen

AU - Obach, R. Scott

AU - O’ Brien, Matthew N.

AU - O’Connor, Rebecca

AU - Ogilvie, Kevin

AU - Owen, Dafydd

AU - Pettersson, Martin

AU - Reese, Matthew R.

AU - Rogers, Thomas F.

AU - Rosales, Romel

AU - Rossulek, Michelle I.

AU - Sathish, Jean G.

AU - Shirai, Norimitsu

AU - Steppan, Claire

AU - Ticehurst, Martyn

AU - Updyke, Lawrence W.

AU - Weston, Stuart

AU - Zhu, Yuao

AU - White, Kris M.

AU - García-Sastre, Adolfo

AU - Wang, Jun

AU - Chatterjee, Arnab K.

AU - Mesecar, Andrew D.

AU - Frieman, Matthew B.

AU - Anderson, Annaliesa S.

AU - Allerton, Charlotte

N1 - Funding Information: The authors would like to thank Sarah Lazzaro, Sumathy Mathialagan, Sangwoo Ryu, Mark West and Emi Yamaguchi (Pfizer) for the transporter inhibition studies. Angela Doran, Chad Limanni, Amanda Plante and Jocelyn Rosado for their in vivo and PK study support (Pfizer). Marcus Ewing (Pfizer) and Gail Johnson (Pfizer) for preformulation studies. Li Hao (Pfizer) for sequence analysis support. Shinji Yamazaki (Pfizer) for PBPK modeling simulations. Daniel Lettiere, Michael Homiski, Michelle Kenyon, Asser Bas-syouni, Declan Flynn, William Reagan, Victoria Markiewicz and Stephen Jenkinson for overseeing safety studies, and William Reagan, for expert clinical pathology and pathology support for the toxicology studies. Deli Huang for supplying the HeLa-ACE2 stably transfected cell line. R. Albert for support with the BSL3 facility and procedures at the Icahn School of Medicine at Mount Sinai, New York. Devendra Rai (Pfizer) and Charles Tan (Pfizer) for their help with the editorial process and statistics. Andrew Mesecar acknowledges partial support for this project from federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Contract No. HHSN272201700060C. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The study performed in Dr. Jun Wang’s laboratory is partially supported by NIH grant (AI147325) and the Young Investigator Award grant from the Arizona Biomedical Research Centre (ADHS18-198859). Scripps work was supported by a grant from the Bill & Melinda Gates Foundation #OPP1107194, and the Scripps Family Impact Fund of the Miramar Charitable Foundation (MCF) MBF is supported by NIH grants R21AI158134, R01 AI148166, R21AI153480, HHSN272201400007C and 75N93019C00051. A DARPA subcontract #HR0011-20-2-0040, BARDA contract #ASPR-20-01495, and Bill and Melinda Gates Foundation grants # INV-006099 and INV-016638. This research was also partly funded by CRIP (Center for Research for Influenza Pathogenesis), a NIAID supported Center of Excellence for Influenza Research and Surveillance (CEIRS, contract # HHSN272201400008C), by DARPA grant HR0011-19-2-0020, by supplements to NIAID grant U19AI142733 U19AI135972 and DoD grant W81XWH-20-1-0270, and by the generous support of the JPB Foundation, the Open Philanthropy Project (research grant 2020-215611 (5384)), and anonymous donors to AG-S. Funding Information: A.D.M. and affiliates (B.J.A., E.K.L.) have a sponsored program contract with Pfizer to test compounds for inhibition of coronavirus proteases. J.W. and affiliate (C.M.) have a sponsored research agreement with Pfizer to test compounds for inhibition of coronavirus proteases. The Frieman Laboratory (M.B.F., H.H., R.E.Ha., J.L., M.E.M., S.W.) was funded by Pfizer for the work in this manuscript. The García-Sastre Laboratory (AGS, KMW, RR) has received research support from Pfizer, Senhwa Biosciences, Accurius, Avimex, 7Hills Pharma, Pharmamar, Blade therapeutics, Dynavax, Kenall Manufacturing, ImmunityBio and Nanocomposix; and AG-S has consulting agreements for the following companies involving cash and/or stock: Vivaldi Biosciences, Contrafect, 7Hills Pharma, Avimex, Valneva, Accurius, and Esperovax. AG-S is inventor in patent applications on SARS-CoV-2 antivirals owned by Icahn School of Medicine at Mount Sinai. B.B., R.M.J., D.A., L.A., J.B., H.E., J.H., R.Ho., E.P.K., R.K., E.K., L.L., J.R.L., S.A.L., S.W.M., S.N., R.S.O., M.N.O., R.O., K.O., D.O., M.P., M.R.R., M.I.R., J.G.S., N.S., C.S., M.T., L.W.U., Y.Z., A.A., and C.A., at the time of their contributions were employees and may have stock in Pfizer. A.K.C., M.A.B., N.B., E.C., M.G.K., T.F.R. have no competing interest to report. Publisher Copyright: © 2021, The Author(s).

PY - 2021/12

Y1 - 2021/12

N2 - COVID-19 caused by the SARS-CoV-2 virus has become a global pandemic. 3CL protease is a virally encoded protein that is essential across a broad spectrum of coronaviruses with no close human analogs. PF-00835231, a 3CL protease inhibitor, has exhibited potent in vitro antiviral activity against SARS-CoV-2 as a single agent. Here we report, the design and characterization of a phosphate prodrug PF-07304814 to enable the delivery and projected sustained systemic exposure in human of PF-00835231 to inhibit coronavirus family 3CL protease activity with selectivity over human host protease targets. Furthermore, we show that PF-00835231 has additive/synergistic activity in combination with remdesivir. We present the ADME, safety, in vitro, and in vivo antiviral activity data that supports the clinical evaluation of PF-07304814 as a potential COVID-19 treatment.

AB - COVID-19 caused by the SARS-CoV-2 virus has become a global pandemic. 3CL protease is a virally encoded protein that is essential across a broad spectrum of coronaviruses with no close human analogs. PF-00835231, a 3CL protease inhibitor, has exhibited potent in vitro antiviral activity against SARS-CoV-2 as a single agent. Here we report, the design and characterization of a phosphate prodrug PF-07304814 to enable the delivery and projected sustained systemic exposure in human of PF-00835231 to inhibit coronavirus family 3CL protease activity with selectivity over human host protease targets. Furthermore, we show that PF-00835231 has additive/synergistic activity in combination with remdesivir. We present the ADME, safety, in vitro, and in vivo antiviral activity data that supports the clinical evaluation of PF-07304814 as a potential COVID-19 treatment.

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UR - http://www.scopus.com/inward/citedby.url?scp=85117608948&partnerID=8YFLogxK

U2 - 10.1038/s41467-021-26239-2

DO - 10.1038/s41467-021-26239-2

M3 - Article

C2 - 34663813

AN - SCOPUS:85117608948

VL - 12

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 6055

ER -

Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19

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