VPS34 inhibitors as SARS-CoV-2 antivirals

总结 SARS-CoV-2是一种β冠状病毒属，是一种有包膜的正义RNA病毒， 对于当前的流行病。由于其对社会和人类健康的深刻影响， 迫切需要了解SARS-CoV-2复制的要求，并确定治疗方法， 战略布局将开发用于其他目的的药物重新用于其他目的可能会提供一种治疗方法的捷径。 发展使用已知靶向特异性宿主因子的化合物也可以阐明关键的 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。CoV感染最显著的特征之一是 建立膜相关的复制细胞器，作为病毒的主要位点 RNA合成。这些膜细胞器的起源还不完全清楚。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，我们询问SARS-CoV-2是否易受脂质代谢调节剂的影响， 评估病毒对VPS 34的VPS 34抑制剂的敏感性，VPS 34是一种脂质激酶， 自噬和内体运输; Triacsin C，一种长链脂肪酰辅酶A抑制剂 脂肪酸合成酶（ACSL）和奥利司他，脂肪酸合成酶（Festival）的抑制剂。我们的初步数据 表明VPS 34抑制剂有效抑制SARS-CoV-2复制，而FDA- 批准的不同类型的PI 3 K抑制剂对复制的影响最小。目标函数 ACSL也会损害SARS-CoV-2的复制。这些数据表明，VPS 34、ACSL和 FGFs在复制中心的形成和病毒生长中起重要作用，并提示这些 酶作为治疗靶点。我们将检验VPS 34、ACSL和FRESIST是 通过评估这些小分子抑制剂对SARS-Cov-2感染至关重要 酶，并通过测量SARS-CoV-2在基因敲除或敲除这些酶中的复制， 宿主酶我们将定义抑制机制并测试生成的假设 膜相关的病毒复制中心将被破坏。最后，我们将评估 VSP 34抑制剂PIK-III和奥利司他全身给药在SARS-coV-2中的体内疗效 感染的仓鼠，以评估脂质代谢抑制剂的治疗潜力。

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.