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

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：开发 ALG-097431 作为广谱 SARS-CoV-2 3CLpro 抑制剂 用小分子药物抑制病毒蛋白酶是一种被证明是成功的治疗方法， 目前用于治疗HCV和HIV感染。 SARS-CoV-2 主要的 3-胰凝乳蛋白酶样半胱氨酸 蛋白酶（Mpro、3CLpro）代表了另一种有前景的病毒蛋白酶靶标，用于无需人类的抗病毒治疗 同系物。 3CLpro 小分子共价结合抑制剂的发现和开发是 本计划的重点。与我们在药物设计和发现中心 (CD3) 的合作者一起， 在比利时鲁汶大学雷加研究所，我们开发了多种生化和细胞检测方法 分析化合物并获得 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 鉴定了一系列高选择性和有效的 3CLpro 抑制剂。抑制 SARS-CoV-2 的复制 这些化合物在表达人 ACE-2 受体的 A549 细胞中得到证实 表现出低纳摩尔 EC50 值。在 A549 细胞中，浓度高达 100μM。这些化合物的活性扩展到其他人类冠状病毒，例如α-冠状病毒 229E 和 β-冠状病毒 OC43，表现出广谱抗冠状病毒活性。代表 化合物 ALG-097111 被纳入体内功效模型，其中仓鼠受到 SARS-CoV-2。感染后两天，RNA 数量显着减少 3.5 log10（RNA 拷贝数/毫克） 观察到病毒 RNA 拷贝数和肺部感染性病毒滴度降低 3.7 log10 (TCID50/mg) 经过 ALG-097111 处理的动物。进一步的化合物优化导致了 ALG-097431 的开发。 与 3CLpro 酶相比，该化合物的 IC50 为 4 nM，EC50 为 39 nM（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 在初级中表现出有效的活性 人气道上皮细胞，气液界面 3D 培养系统 (EC90 = 12 nM)。该化合物表明 干净的体外 ADME-tox 特征，并在大鼠和狗中显示出良好的口服生物利用度。人体有效剂量- 预测表明每日两次给药 250 – 600 mg 剂量的潜力，无需联合用药 与利托那韦一起给药。该项目的目标是将 ALG-097431 推进人体临床试验，同时 同时优化更多化合物以增强抗病毒活性和 PK 特性。最初这将 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。遵循探索性安全药理学研究、GMP 药物合成、制剂开发和 将启动生物分析方法的验证。在开始临床研究之前，潜在的毒性 ALG-097431 的成分将在根据 GLP 法规进行的多项体外和体内研究中进行评估。