Development of ALG-097431 as a broad-spectrum 3CL pro inhibitor

开发 ALG-097431 作为广谱 3CL 前体抑制剂

• 批准号: 10513921
• 负责人: Julian A Symons
• 金额: $ 422.8万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10513921
• 关键词: 2019-nCoV; A549; ACE2; Adult; Air; Animals; Antiviral Agents; Antiviral Therapy; Appearance; Belgium; Binding; Biochemical; Biological Assay; Biological Availability; CD3 Antigens; Canis familiaris; Cardiovascular system; Caspase; Cells; Chemistry; Clinical Research; Clinical Trials; Coronavirus; Crystallization; Development; Dose; Drug Design; Enzymes; Epithelial Cells; Excretory function; Exhibits; Formulation; Goals; HIV Infections; Hamsters; Hepatitis C; Hepatitis C Therapy; Homologous Gene; Human; In Vitro; Infection; Institutes; Ion Channel; Ions; Liquid substance; Lung; Metabolism; Methods; Modeling; Monitor; Neuraxis; Oral; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Pharmacology Study; Phosphotransferases; Phototoxicity; Process; Property; RNA; Rattus; Regulation; Respiratory System; Ritonavir; SARS-CoV-2 B.1.1.7; SARS-CoV-2 infection; Safety; Series; Solubility; Sprague-Dawley Rats; Structure; System; Therapeutic; Toxic effect; Toxicology; Validation; Viral; Virus; absorption; airway epithelium; base; betacoronavirus; chemical property; chymotrypsin; cytotoxicity; drug discovery; drug synthesis; enantiomer; genotoxicity; good laboratory practice; human coronavirus; improved; in vivo; indexing; inhibitor; metropolitan; nanomolar; pharmacokinetics and pharmacodynamics; physical property; programs; receptor; safety assessment; safety study; scale up; screening panel; small molecule; three dimensional cell culture; viral RNA; 2019-nCoV; A549; ACE2; Adult; Air; Animals; Antiviral Agents; Antiviral Therapy; Appearance; Belgium; Binding; Biochemical; Biological Assay; Biological Availability; CD3 Antigens; Canis familiaris; Cardiovascular system; Caspase; Cells; Chemistry; Clinical Research; Clinical Trials; Coronavirus; Crystallization; Development; Dose; Drug Design; Enzymes; Epithelial Cells; Excretory function; Exhibits; Formulation; Goals; HIV Infections; Hamsters; Hepatitis C; Hepatitis C Therapy; Homologous Gene; Human; In Vitro; Infection; Institutes; Ion Channel; Ions; Liquid substance; Lung; Metabolism; Methods; Modeling; Monitor; Neuraxis; Oral; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Pharmacology Study; Phosphotransferases; Phototoxicity; Process; Property; RNA; Rattus; Regulation; Respiratory System; Ritonavir; SARS-CoV-2 B.1.1.7; SARS-CoV-2 infection; Safety; Series; Solubility; Sprague-Dawley Rats; Structure; System; Therapeutic; Toxic effect; Toxicology; Validation; Viral; Virus; absorption; airway epithelium; base; betacoronavirus; chemical property; chymotrypsin; cytotoxicity; drug discovery; drug synthesis; enantiomer; genotoxicity; good laboratory practice; human coronavirus; improved; in vivo; indexing; inhibitor; metropolitan; nanomolar; pharmacokinetics and pharmacodynamics; physical property; programs; receptor; safety assessment; safety study; scale up; screening panel; small molecule; three dimensional cell culture; viral RNA

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.

项目摘要/摘要：ALG-097431的开发为广谱SARS-COV-2 3Clpro 抑制剂 用小分子药物抑制病毒蛋白酶是一种成功的热方法，是 目前用于治疗HCV和HIV感染。 SARS-COV-2主，三键型类半胱氨酸 蛋白酶（MPRO，3CLPRO）代表了没有人类的另一个有前途的病毒蛋白酶靶标的抗病毒药疗法 同源物。 3clpro的小分子的发现和开发，共价结合抑制剂是 该程序的重​​点。与我们在药物设计与发现中心合作者（CD3）和 比利时Ku Leuven的Rega Institute，我们已经开发了多种生化和基于细胞的测定法 曲线化合物，并可以访问SARS-COV-2感染的仓鼠模型。通过基于结构的 优化，结合选择性和SARS-COV-2 3Clpro生化酶测定，我们有 确定了一系列高度选择性和潜在的3CLPRO抑制剂。 SARS-COV-2的复制抑制 这些化合物在表达具有化合物的人ACE-2受体的A549细胞中得到了证实 证明低纳摩尔EC50值。在A549细胞中未观察到细胞毒性，浓度为 100μm。这些化合物的活性扩展到其他人类冠状病毒，例如α-核可纳病毒 229E和β-核纳病毒OC43，表明宽光谱抗核心管活性。代表 ALG-097111化合物被带入一个体内效率模型，在该模型中，仓鼠受到挑战 SARS-CoV-2。感染后两天，鲁棒且有效的3.5 log10（RNA副本/mg）还原 在肺部传染病滴度中的病毒RNA副本和3.7 log10（TCID50/mg）降低 ALG-097111处理的动物。进一步的复合优化导致了ALG-097431的发展。 该化合物的IC50为4 nm，而3Clpro酶和39 nm的EC50（选择性指数 > 1200）在A549-ACE2/B.1.1.7基于细胞的测定中。此外，ALG-097431在主要的 人气道上皮细胞，空气界面3D培养系统（EC90 = 12 nm）。该化合物演示 干净的体外ADME-TOX曲线，并显示出大鼠和狗的口服生物利用度良好。人类有效剂量 - 项目表明，每天两次给药的可能性，不需要250 - 600毫克的剂量 利托纳维尔的管理。该项目的目标是将ALG-097431推进到人类的临床试验中 类似地优化了增强抗病毒活性和PK特性的其他化合物。最初这将是 涉及建立PK/PD相关性并进行进一步的选择性分析和更广泛的PK 分析。探索性安全药理学研究，GMP药物合成，配方奶粉开发和 将启动生物分析方法的验证。在临床研究计划之前，潜在的毒性 ALG-097431将在根据GLP法规进行的多个体外和体内研究中进行评估。