背景:在寻找抗COVID-19疗法时,1,2,3,4,6-五-O-没食子酰-βD-吡喃葡萄糖苷,一种从许多传统草药中分离出来的天然多酚化合物,已被报道为RBD-ACE2结合抑制剂和靶向SARSCoV-2的主要蛋白酶和RNA依赖性RNA聚合酶的广谱抗冠状病毒抑制剂。为了促进1,2,3,4,6-五-O-没食子酰-β-D-吡喃葡萄糖苷的构效关系研究,我们描述了它的化学合成和表征,以及它对SARS-CoV-2与宿主ACE2受体相互作用的活性。
方法:由3,4,5-三苄氧苯甲酸和β-D-吡喃葡萄糖苷通过两个定量步骤合成了1,2,3,4,6-五烷基-O-没食子酰基-β-D-吡喃葡萄糖苷:DCC介导的酯化和钯催化的去苄基化。使用SARS-CoV-2尖峰三聚体(S1S2)ACE2抑制剂筛选比色测定试剂盒评估合成的分子,SARS-CoV2尖峰S1RBDACE2抑制剂筛选比色测定试剂盒,和使用Spike(SARS-CoV-2)假型慢病毒的细胞中和测定法,ACE2-HEK293重组细胞系。
结果:化学合成的产物阻断了SARSCoV-2的尖峰三聚体与人ACE2受体的结合,IC50=22±2µM。它还以IC50=27±3µM阻断ACE2:尖峰RBD结合。重要的是,它抑制了SARS2-CoV2-Spike假型化慢病毒对ACE2HEK293细胞系的感染性,IC50=20±2µM。
结论:总体而言,化学合成的1,2,3,4,6-五-O-没食子酰-β-D-吡喃葡萄糖苷是开发抗SARS-CoV-2疗法的先导分子,该疗法通过阻断病毒进入宿主细胞来阻断病毒感染的初始阶段。
BACKGROUND: In the search for anti-COVID-19 therapy, 1,2,3,4,6-pentakis-O-galloyl-βD-glucopyranoside, a natural polyphenolic compound isolated from many traditional medicinal herbs, has been reported as an RBD-ACE2 binding inhibitor and as a broad-spectrum anticoronaviral inhibitor targeting the main protease and RNA-dependent RNA polymerase of SARSCoV-2. To facilitate the structure-activity relationship studies of 1,2,3,4,6-pentakis-O-galloyl-β-Dglucopyranoside, we describe its chemical synthesis and characterization, as well as its activity towards the SARS-CoV-2 spike interaction with host ACE2 receptor.
METHODS: 1,2,3,4,6-Pentakis-O-galloyl-β-D-glucopyranoside was synthesized in two quantitative steps from 3,4,5-tribenzyloxybenzoic acid and β-D-glucopyranoside: DCC-mediated esterification and palladium-catalyzed per-debenzylation. The synthesized molecule was evaluated using a SARS-CoV-2 spike trimer (S1 + S2) ACE2 inhibitor screening colorimetric assay kit, SARS-CoV2 spike S1 RBD ACE2 inhibitor screening colorimetric assay kit, and a cellular neutralization assay using the Spike (SARS-CoV-2) Pseudotyped Lentivirus, ACE2-HEK293 recombinant cell line.
RESULTS: The chemically synthesized product blocked the binding of the spike trimer of SARSCoV-2 to the human ACE2 receptor with IC50=22±2 µM. It also blocked ACE2:spike RBD binding with IC50=27±3 µM. Importantly, it inhibited the infectivity of SARS2-CoV2-Spike pseudotyped lentivirus on the ACE2 HEK293 cell line with IC50=20±2 µM.
CONCLUSIONS: Overall, the chemically synthesized 1,2,3,4,6-pentakis-O-galloyl-β-D-glucopyranoside represents a lead molecule to develop anti-SARS-CoV-2 therapies that block the initial stage of the viral infection by blocking the virus entry to the host cell.