作者: Gunesch, Antonia P ; Pietschmann, Thomas ; Milke, Vanessa ; Kühl, David ; Haid, Sibylle ; Schulz, Thomas F ; Lasswitz, Lisa ; Brown, Richard J P ; Köhler, Natalie M ; Sake, Svenja M ; Nguyen, Tu-Trinh ; Brönstrup, Mark ; Wiegmann, Bettina ; Vieyres, Gabrielle ; Gerold, Gisa ; Rückert, Jessica ; Bilitewski, Ursula ; Matthaei, Alina ; Winkler, Melina ; Brogden, Graham ; Göhl, Matthias ; Zapatero-Belinchón, Francisco J

ABSTRACT Libraries composed of licensed drugs represent a vast repertoire of molecules modulating physiological processes in humans, providing unique opportunities for the discovery of host-targeting antivirals. We screened the Repurposing, Focused Rescue, and Accelerated Medchem (ReFRAME) repurposing library with approximately 12,000 molecules for broad-spectrum coronavirus antivirals and discovered 134 compounds inhibiting an alphacoronavirus and mapping to 58 molecular target categories. Dominant targets included the 5-hydroxytryptamine receptor, the dopamine receptor, and cyclin-dependent kinases. Gene knock-out of the drugs’ host targets including cathepsin B and L (CTSB/L; VBY-825), the aryl hydrocarbon receptor (AHR; Phortress), the farnesyl-diphosphate farnesyltransferase 1 (FDFT1; P-3622), and the kelch-like ECH-associated protein 1 (KEAP1; Omaveloxolone), significantly modulated HCoV-229E infection, providing evidence that these compounds inhibited the virus through acting on their respective host targets. Counter-screening of all 134 primary compound candidates with SARS-CoV-2 and validation in primary cells identified Phortress, an AHR activating ligand, P-3622-targeting FDFT1, and Omaveloxolone, which activates the NFE2-like bZIP transcription factor 2 (NFE2L2) by liberating it from its endogenous inhibitor KEAP1, as antiviral candidates for both an Alpha - and a Betacoronavirus . This study provides an overview of HCoV-229E repurposing candidates and reveals novel potentially druggable viral host dependency factors hijacked by diverse coronaviruses.

2023-09-20·Microbiology spectrum

A novel in vitro system of supported planar endosomal membranes (SPEMs) reveals an enhancing role for cathepsin B in the final stage of Ebola virus fusion and entry.

Article

作者: Kiessling, Volker ; Tamm, Lukas K ; White, Judith M ; Odongo, Laura ; Habtegebrael, Betelihem H

Ebola virus (EBOV) causes a hemorrhagic fever with fatality rates up to 90%. The EBOV entry process is complex and incompletely understood. Following attachment to host cells, EBOV is trafficked to late endosomes/lysosomes where its glycoprotein (GP) is processed to a 19-kDa form, which binds to the EBOV intracellular receptor Niemann-Pick type C1. We previously showed that the cathepsin protease inhibitor, E-64d, blocks infection by pseudovirus particles bearing 19-kDa GP, suggesting that further cathepsin action is needed to trigger fusion. This, however, has not been demonstrated directly. Since 19-kDa Ebola GP fusion occurs in late endosomes, we devised a system in which enriched late endosomes are used to prepare supported planar endosomal membranes (SPEMs), and fusion of fluorescent (pseudo)virus particles is monitored by total internal reflection fluorescence microscopy. We validated the system by demonstrating the pH dependencies of influenza virus hemagglutinin (HA)-mediated and Lassa virus (LASV) GP-mediated fusion. Using SPEMs, we showed that fusion mediated by 19-kDa Ebola GP is dependent on low pH, enhanced by Ca²⁺, and augmented by the addition of cathepsins. Subsequently, we found that E-64d inhibits full fusion, but not lipid mixing, mediated by 19-kDa GP, which we corroborated with the reversible cathepsin inhibitor VBY-825. Hence, we provide both gain- and loss-of-function evidence that further cathepsin action enhances the fusion activity of 19-kDa Ebola GP. In addition to providing new insights into how Ebola GP mediates fusion, the approach we developed employing SPEMs can now be broadly used for studies of virus and toxin entry through endosomes. IMPORTANCE Ebola virus is the causative agent of Ebola virus disease, which is severe and frequently lethal. EBOV gains entry into cells via late endosomes/lysosomes. The events immediately preceding fusion of the viral and endosomal membranes are incompletely understood. In this study, we report a novel in vitro system for studying virus fusion with endosomal membranes. We validated the system by demonstrating the low pH dependencies of influenza and Lassa virus fusion. Moreover, we show that further cathepsin B action enhances the fusion activity of the primed Ebola virus glycoprotein. Finally, this model endosomal membrane system should be useful in studying the mechanisms of bilayer breaching by other enveloped viruses, by non-enveloped viruses, and by acid-activated bacterial toxins.

2020-10-01·Nature1区 · 综合性期刊

Discovery of SARS-CoV-2 antiviral drugs through large-scale compound repurposing

1区 · 综合性期刊

Article

作者: Burgstaller-Muehlbacher, Sebastian ; Mesecar, Andrew D ; Cheng, Kuoyuan ; Liu, Wen-Chun ; White, Kris M ; García-Sastre, Adolfo ; Herbert, Kristina M ; Ruppin, Eytan ; Zwaka, Thomas P ; Poon, Vincent Kwok-Man ; Schotsaert, Michael ; Nguyen, Tu-Trinh H ; Martinez-Sobrido, Luis ; Matsunaga, Naoko ; De Jesus, Paul D ; Su, Andrew I ; Choi, Angela ; Albrecht, Randy ; Benner, Christopher ; Schultz, Peter G ; Pu, Yuan ; Sun, Ren ; Rubanov, Andrey ; Sit, Ko-Yung ; Chanda, Sumit K ; Glynne, Richard J ; Rathnasinghe, Raveen ; Hull, Mitchell V ; Chang, Max W ; Pache, Lars ; Lendy, Emma K ; Chan, Jasper Fuk-Woo ; Chatterjee, Arnab K ; Yuan, Shuofeng ; Miorin, Lisa ; Yin, Xin ; Cao, Jianli ; Martin-Sancho, Laura ; Chapman, Mackenzie E ; Dejosez, Marion ; Johnson, Jeffrey R ; Nguyen, Courtney ; Teriete, Peter ; Riva, Laura ; Yuen, Kwok-Yung

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 2019 has triggered an ongoing global pandemic of the severe pneumonia-like disease coronavirus disease 2019 (COVID-19)¹. The development of a vaccine is likely to take at least 12-18 months, and the typical timeline for approval of a new antiviral therapeutic agent can exceed 10 years. Thus, repurposing of known drugs could substantially accelerate the deployment of new therapies for COVID-19. Here we profiled a library of drugs encompassing approximately 12,000 clinical-stage or Food and Drug Administration (FDA)-approved small molecules to identify candidate therapeutic drugs for COVID-19. We report the identification of 100 molecules that inhibit viral replication of SARS-CoV-2, including 21 drugs that exhibit dose-response relationships. Of these, thirteen were found to harbour effective concentrations commensurate with probable achievable therapeutic doses in patients, including the PIKfyve kinase inhibitor apilimod^2-4 and the cysteine protease inhibitors MDL-28170, Z LVG CHN2, VBY-825 and ONO 5334. Notably, MDL-28170, ONO 5334 and apilimod were found to antagonize viral replication in human pneumocyte-like cells derived from induced pluripotent stem cells, and apilimod also demonstrated antiviral efficacy in a primary human lung explant model. Since most of the molecules identified in this study have already advanced into the clinic, their known pharmacological and human safety profiles will enable accelerated preclinical and clinical evaluation of these drugs for the treatment of COVID-19.

100 项与 VBY-825 相关的药物交易

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