VPS34 inhibitors as SARS-CoV-2 antivirals

VPS34 抑制剂作为 SARS-CoV-2 抗病毒药物

• 批准号: 10534720
• 负责人: Christopher F Basler
• 金额: $ 41.59万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10534720
• 关键词: VPS34; inhibitors; SARS; CoV; antivirals

Summary SARS-CoV-2, a Betacoronavirus genus, is an enveloped positive-sense, RNA virus responsible for a current pandemic. Because of its profound impact on society and human health there is an urgent need to understand SARS-CoV-2 replication requirements and to identify therapeutic strategies. Repurposing drugs developed for other purposes may provide a shortcut to therapeutic development. The use of compounds known to target specific host factors may also elucidate key pathways needed for virus replication. Coronavirus (CoV) replication involves multiple critical interactions with host cell membranes. One of the most striking features of CoV infection is the establishment of membrane-associated replication organelles that serve as the main sites of viral RNA synthesis. The origin of these membrane organelles is incompletely understood. Because the specific host pathways required for SARS-CoV-2 replication organelle formation are not defined, we asked whether SARS-CoV-2 is susceptible to modulators of lipid metabolism by assessing the sensitivity of the virus to VPS34 inhibitors of VPS34, a lipid kinase required for autophagy and endosomal trafficking; Triacsin C, an inhibitor of long chain fatty acyl CoA synthetase (ACSL) and Orlistat, an inhibitor of fatty acid synthase (FASN). Our preliminary data indicate that inhibitors of VPS34 potently inhibited SARS-CoV-2 replication, whereas an FDA- approved inhibitor of a different class of PI3K had minimal effect on replication. Targeting FASN and ACSL also impairs SARS-CoV-2 replication. These data suggest that VPS34, ACSL and FASN play important roles in replication center formation and virus growth and suggest these enzymes as therapeutic targets. We will test the hypothesis that VPS34, ACSL and FASN are critical for SARS-Cov-2 infection by evaluating additional small molecule inhibitors of these enzymes and by measuring SARS-CoV-2 replication in genetic knockdowns or knockouts of these host enzymes. We will define mechanisms of inhibition and test the hypothesis that generation of membrane-associated viral replication centers will be disrupted. Finally, we will assess the in vivo efficacy of VSP34 inhibitor PIK-III and systemically administered Orlistat in SARS-coV-2- infected hamsters to evaluate the therapeutic potential of inhibitors of lipid metabolism.

概括 SARS-CoV-2 是乙型冠状病毒属，是一种有包膜的正义 RNA 病毒，负责 针对当前的大流行。由于其对社会和人类健康的深远影响， 迫切需要了解 SARS-CoV-2 复制要求并确定治疗方法 策略。将药物重新用于其他目的可能会提供一条治疗的捷径 发展。使用已知针对特定宿主因子的化合物也可能阐明关键的 病毒复制所需的途径。冠状病毒（CoV）复制涉及多个关键 与宿主细胞膜的相互作用。冠状病毒感染最显着的特征之一是 建立作为病毒主要位点的膜相关复制细胞器 RNA合成。这些膜细胞器的起源尚不完全清楚。因为 SARS-CoV-2 复制细胞器形成所需的特定宿主途径并不 定义，我们通过以下方式询问 SARS-CoV-2 是否对脂质代谢调节剂敏感 评估病毒对 VPS34 抑制剂的敏感性，VPS34 是一种脂质激酶， 自噬和内体运输； Triacsin C，长链脂肪酰辅酶 A 抑制剂 合成酶（ACSL）和奥利司他（一种脂肪酸合成酶（FASN）抑制剂）。我们的初步数据 表明 VPS34 抑制剂可有效抑制 SARS-CoV-2 复制，而 FDA- 已批准的不同类别 PI3K 抑制剂对复制的影响极小。针对 FASN ACSL 还会损害 SARS-CoV-2 复制。这些数据表明 VPS34、ACSL 和 FASN 在复制中心形成和病毒生长中发挥重要作用，并表明这些 酶作为治疗靶点。我们将测试 VPS34、ACSL 和 FASN 的假设 通过评估这些药物的其他小分子抑制剂，对 SARS-Cov-2 感染至关重要 酶并通过基因敲除或基因敲除来测量 SARS-CoV-2 的复制 宿主酶。我们将定义抑制机制并测试一代的假设 膜相关病毒复制中心的数量将被破坏。最后，我们将评估 VSP34 抑制剂 PIK-III 和全身给药奥利司他在 SARS-coV-2- 中的体内疗效 感染仓鼠以评估脂质代谢抑制剂的治疗潜力。