• Discovery of SARS-CoV-2 Papain-like Protease Inhibitors through a Combination of High-Throughput Screening and a FlipGFP-Based Reporter Assay

      Ma, C.; Sacco, M.D.; Xia, Z.; Lambrinidis, G.; Townsend, J.A.; Hu, Y.; Meng, X.; Szeto, T.; Ba, M.; Zhang, X.; et al. (American Chemical Society, 2021)
      The papain-like protease (PLpro) of SARS-CoV-2 is a validated antiviral drug target. Through a fluorescence resonance energy transfer-based high-throughput screening and subsequent lead optimization, we identified several PLpro inhibitors including Jun9-72-2 and Jun9-75-4 with improved enzymatic inhibition and antiviral activity compared to GRL0617, which was reported as a SARS-CoV PLpro inhibitor. Significantly, we developed a cell-based FlipGFP assay that can be applied to predict the cellular antiviral activity of PLpro inhibitors in the BSL-2 setting. X-ray crystal structure of PLpro in complex with GRL0617 showed that binding of GRL0617 to SARS-CoV-2 induced a conformational change in the BL2 loop to a more closed conformation. Molecular dynamics simulations showed that Jun9-72-2 and Jun9-75-4 engaged in more extensive interactions than GRL0617. Overall, the PLpro inhibitors identified in this study represent promising candidates for further development as SARS-CoV-2 antivirals, and the FlipGFP-PLpro assay is a suitable surrogate for screening PLpro inhibitors in the BSL-2 setting. © 2021 The Authors. Published by American Chemical Society.
    • Flaggite, Pb4Cu2+4Te6+2(SO4)2O11(OH)2(H2O), a new mineral with stair-step-like HCP layers from Tombstone, Arizona, USA

      Kampf, A.R.; Mills, S.J.; Celestian, A.J.; Ma, C.; Yang, H.; Thorne, B.; Department of Geosciences, University of Arizona (Cambridge University Press, 2022)
      The new mineral flaggite (IMA2021–044), Pb4Cu2+4Te6+2(SO4)2O11(OH)2(H2O), occurs at the Grand Central mine in the Tombstone district, Cochise County, Arizona, USA, in cavities in quartz matrix in association with alunite, backite, cerussite, jarosite and rodalquilarite. Flaggite crystals are lime-green to yellow-green tablets, up to 0.5 mm across. The mineral has a very pale green streak and adamantine lustre. It is brittle with irregular fracture and a Mohs hardness of about 3. It has one excellent cleavage on {010}. The calculated density is 6.137 g cm–3. Optically, the mineral is biaxial (+) with α = 1.95(1), β = 1.96(1), γ = 2.00(1) (white light); 2V = 54(2)°; pleochroism: X green, Y light yellow green, Z nearly colorless; X > Y > Z. The Raman spectrum exhibits bands consistent with TeO6and SO4. Electron microprobe analysis provided the empirical formula Pb3.88Cu2+3.89Te6+2.08(SO4)2O11(OH)2(H2O) (–0.03 H). Flaggite is triclinic, P1, a = 9.5610(2), b = 9.9755(2), c = 10.4449(3) Å, α = 74.884(1), β = 89.994(1), γ = 78.219(1)°, V = 939.97(4) Å3and Z = 2. The structure of flaggite (R1= 0.0342 for 5936 I > 2σI) contains hexagonal-close-packed, stair-step-like layers comprising TeO6octahedra and Jahn-Teller distorted CuO6octahedra. The layers in the structure of flaggite are very similar to those in bairdite, timroseite and paratimroseite. © 2022 Cambridge University Press. All rights reserved.
    • Preclinical characterization of an intravenous coronavirus 3CL protease inhibitor for the potential treatment of COVID19

      Boras, B.; Jones, R.M.; Anson, B.J.; Arenson, D.; Aschenbrenner, L.; Bakowski, M.A.; Beutler, N.; Binder, J.; Chen, E.; Eng, H.; et al. (Nature Research, 2021)
      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. © 2021, The Author(s).
    • Rational design of a deuterium-containing M2-S31N channel blocker UAWJ280 with in vivo antiviral efficacy against both oseltamivir sensitive and -resistant influenza A viruses

      Cáceres, C.J.; Hu, Y.; Cárdenas-García, S.; Wu, X.; Tan, H.; Carnaccini, S.; Gay, L.C.; Geiger, G.; Ma, C.; Zhang, Q.-Y.; et al. (Taylor and Francis Ltd., 2021)
      Seasonal influenza A virus (IAV) infections are among the most important global health problems. FDA-approved antiviral therapies against IAV include neuraminidase inhibitors, M2 inhibitors, and polymerase inhibitor baloxavir. Resistance against adamantanes (amantadine and rimantadine) is widespread as virtually all IAV strains currently circulating in the human population are resistant to adamantanes through the acquisition of the S31N mutation. The neuraminidase inhibitor-resistant strains also contain the M2-S31N mutant, suggesting M2-S31N is a high-profile antiviral drug target. Here we report the development of a novel deuterium-containing M2-S31N inhibitor UAWJ280. UAWJ280 had broad-spectrum antiviral activity against both oseltamivir sensitive and -resistant influenza A strains and had a synergistic antiviral effect in combination with oseltamivir in cell culture. In vivo pharmacokinetic (PK) studies demonstrated that UAWJ280 had favourable PK properties. The in vivo mouse model study showed that UAWJ280 was effective alone or in combination with oseltamivir in improving clinical signs and survival after lethal challenge with an oseltamivir sensitive IAV H1N1 strain. Furthermore, UAWJ280 was also able to ameliorate clinical signs and increase survival when mice were challenged with an oseltamivir-resistant IAV H1N1 strain. In conclusion, we show for the first time that the M2-S31N channel blocker UAWJ280 has in vivo antiviral efficacy in mice that are infected with either oseltamivir sensitive or -resistant IAVs, and it has a synergistic antiviral effect with oseltamivir. © 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
    • Structure and inhibition of the SARS-CoV-2 main protease reveal strategy for developing dual inhibitors against Mpro and cathepsin L

      Sacco, M.D.; Ma, C.; Lagarias, P.; Gao, A.; Townsend, J.A.; Meng, X.; Dube, P.; Zhang, X.; Hu, Y.; Kitamura, N.; et al. (American Association for the Advancement of Science, 2020)
      The main protease (Mpro) of SARS-CoV-2 is a key antiviral drug target. While most Mpro inhibitors have a γ-lactam glutamine surrogate at the P1 position, we recently found that several Mpro inhibitors have hydrophobic moieties at the P1 site, including calpain inhibitors II and XII, which are also active against human cathepsin L, a host protease that is important for viral entry. In this study, we solved x-ray crystal structures of Mpro in complex with calpain inhibitors II and XII and three analogs of GC-376. The structure of Mpro with calpain inhibitor II confirmed that the S1 pocket can accommodate a hydrophobic methionine side chain, challenging the idea that a hydrophilic residue is necessary at this position. The structure of calpain inhibitor XII revealed an unexpected, inverted binding pose. Together, the biochemical, computational, structural, and cellular data presented herein provide new directions for the development of dual inhibitors as SARS-CoV-2 antivirals. Copyright © 2020 The Authors, some rights reserved.