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Principal Investigator: Julian A Symons
Organization: HACKENSACK UNIVERSITY MEDICAL CENTER
Fiscal Year: 2022
Award: $4,228,031
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary/Abstract: Development of ALG-097431 as a Broad-Spectrum SARS-CoV-2 3CLpro
Inhibitor
Inhibition of viral proteases with small molecule drugs is a proven successful therapeutic approach and is
currently used for the treatment of HCV and HIV infection. The SARS-CoV-2 main, 3-chymotrypsin-like cysteine
protease (Mpro, 3CLpro) represents another promising viral protease target for antiviral therapy with no human
homolog. The discovery and development of small molecule, covalent binding inhibitors of the 3CLpro is the
focus of this program. Together with our collaborators at the Centre for Drug Design and Discovery (CD3) and
the Rega Institute at KU Leuven, Belgium, we have developed multiple biochemical and cell-based assays to
profile compounds and have access to the hamster model of SARS-CoV-2 infection. Through structure-based
optimization, combined with selectivity and SARS-CoV-2 3CLpro biochemical enzymatic assays, we have
identified a series of highly selective and potent 3CLpro inhibitors. The inhibition of replication of SARS-CoV-2
by these compounds was confirmed in A549 cells expressing the human ACE-2 receptor with compounds
demonstrating low nanomolar EC50 values. No cytotoxicity was observed in A549 cells at concentrations up to
100 μM. The activity of these compounds extended to other human coronaviruses such as the alpha-coronavirus
229E and the beta-coronavirus OC43, demonstrating broad-spectrum anti-coronaviral activity. A representative
compound, ALG-097111, was taken into an in vivo efficacy model in which hamsters were challenged with
SARS-CoV-2. At two days post-infection, a robust and significant 3.5 log10 (RNA copies/mg) reduction of the
viral RNA copies and 3.7 log10 (TCID50/mg) reduction in the infectious virus titers in the lungs was observed in
ALG-097111-treated animals. Further compound optimization has resulted in the development of ALG-097431.
This compound demonstrates an IC50 of 4 nM vs. the 3CLpro enzyme and an EC50 of 39 nM (selectivity index of
>1200) in a A549-ACE2/B.1.1.7 cell-based assay. Further, ALG-097431 exhibits potent activity in a primary
human airway epithelial cell, air-liquid interface 3D-culture system (EC90 = 12 nM). The compound demonstrates
a clean in vitro ADME-tox profile and shows good oral bioavailability in rats and dogs. Human efficacious dose-
projections indicate the potential for twice-daily dosing with a 250 – 600 mg dose without the need for co-
administration with ritonavir. The goals of this project are to advance ALG-097431 into human clinical trials while
simultaneously optimizing further compounds for enhanced antiviral activity and PK properties. Initially this will
involve establishing PK/PD correlations and conducting further selectivity analysis and more extensive PK
analysis. Following exploratory safety pharmacology studies, GMP drug synthesis, formulation development and
validation of bioanalytical methods will be initiated. Prior to the initiation of clinical studies, the potential toxicity
of ALG-097431 will be evaluated in multiple in vitro and in vivo studies conducted under GLP regulations.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><21+ years old><3D cell culture><3D culture><A549><ACE2><Adult><Adult Human><Air><Animals><Antiviral Agents><Antiviral Drugs><Antiviral Therapy><Antivirals><Appearance><Assay><B.1.1.7><Belgium><Binding><Bioassay><Bioavailability><Biochemical><Biologic Assays><Biologic Availability><Biological Assay><Biological Availability><CD3><CD3 Antigens><CD3 Complex><CD3 molecule><CNS Nervous System><COVID-19 infection><COVID-19 virus><COVID19 infection><COVID19 virus><Canine Species><Canis familiaris><Cardiovascular><Cardiovascular Body System><Cardiovascular Organ System><Cardiovascular system><Caspase><Caspase Gene><Cell Body><Cell-Death Protease><Cells><Central Nervous System><Chemistry><Clinical Research><Clinical Study><Clinical Trials><CoV-2><CoV2><Common Rat Strains><Coronaviridae><Coronavirus><Cricetinae><Crystallization><Cysteine Endopeptidases><Cysteine Protease><Cysteine Proteinases><Development><Dogs><Dogs Mammals><Dose><Drug Design><Drug Synthesis and Chemistry><Drugs><Enzyme Gene><Enzymes><Epithelial Cells><Esteroproteases><Excretory function><Exhibits><Formulation><GeneHomolog><Goals><HCV infection><HCV therapy><HCV treatment><HCoV><HIV Infections><HTLV-III Infections><HTLV-III-LAV Infections><Hamsters><Hamsters Mammals><Heart Vascular><Hepatitis C><Hepatitis C Therapeutics><Hepatitis C Therapy><Hepatitis C Virus Treatment><Hepatitis C treatment><Hepatitis C virus infection><Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted><Hepatitus C><Homolog><Homologous Gene><Homologue><Human><Human T-Lymphotropic Virus Type III Infections><ICE-like protease><In Vitro><Infection><Institutes><Intermediary Metabolism><Ion Channel><Ionic Channels><Ions><Kinases><Liquid substance><Lung><Lung Respiratory System><Lytotoxicity><Medication><Membrane Channels><Metabolic Processes><Metabolism><Methods><Modeling><Modern Man><Molecular Interaction><Monitor><Neuraxis><Non-Polyadenylated RNA><OKT3 antigen><Oral><PK/PD><Peptidases><Peptide Hydrolases><Pharmaceutic Preparations><Pharmaceutical Agent><Pharmaceutical Preparations><Pharmaceuticals><Pharmacologic Substance><Pharmacological Study><Pharmacological Substance><Pharmacology><Pharmacology Study><Phosphotransferase Gene><Phosphotransferases><Phototoxicity><Physiologic Availability><Process><Property><Protease Gene><Proteases><Proteinases><Proteolytic Enzymes><Pulmonary Body System><Pulmonary Organ System><RNA><RNA Gene Products><Rat><Rats Mammals><Rattus><Receptor Protein><Regulation><Respiratory Epithelium><Respiratory System><Respiratory Tracts><Respiratory tract structure><Ribonucleic Acid><Ritonavir><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 B.1.1.7><SARS-CoV-2 alpha><SARS-CoV-2 infection><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Safety><Series><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Solubility><Sprague-Dawley Rats><Structure><Structure of respiratory epithelium><System><T3 Antigens><T3 Complex><T3 molecule><Therapeutic><Toxic effect><Toxicities><Toxicology><Transphosphorylases><U.K. variant><UK strain><UK variant><United Kingdom variant><Validation><Viral><Virus><Wuhan coronavirus><absorption><adulthood><airway epithelium><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-viral agents><anti-viral compound><anti-viral drugs><anti-viral medication><anti-viral therapeutic><anti-viral therapy><anti-virals><antiviral compound><antiviral medication><antiviral therapeutic><base><beta CoV><beta coronavirus><betaCoV><betacoronavirus><canine><chemical property><chymotrypsin><circulatory system><corona virus><coronavirus disease 2019 infection><coronavirus disease 2019 virus><coronavirus disease-19 virus><cystein protease><cystein proteinase><cysteine endopeptidase><cytotoxicity><developmental><domestic dog><drug discovery><drug synthesis><drug/agent><enantiomer><excretion><fluid><genotoxicity><good laboratory practice><hCoV19><hep C><hepatitis non A non B><human CoV><human corona virus><human coronavirus><improved><in vivo><indexing><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><inhibitor><liquid><metropolitan><nCoV2><nano-molar><nanomolar><non A, non B hepatitis><non-A, non-B hepatitis><norvir><pharmacokinetics and pharmacodynamics><physical property><programs><pulmonary><receptor><safety assessment><safety study><scale up><screening panel><severe acute respiratory syndrome coronavirus 2 B.1.1.7><small molecule><three dimensional cell culture><viral RNA><viral infectious disease treatment><virus RNA><β CoV><β coronavirus><βCoV>