基于Mpro和PLP筛选潜在抗新型冠状病毒中药研究

新型冠状病毒感染疫情发展迅速,中药在这场防治疫情的战争中发挥着积极作用,已有大量确有疗效的方剂应用于临床。然而由于中药具有多成分、多靶点、多途径协同作用的特点,传统的抗病毒中药筛选研究受限于实验室条件和实验周期,极大地限制了筛选速度。该文基于分子对接进行潜在抗新型冠状病毒中药的筛选,通过获取2019-nCoV冠状病毒3CL水解酶(3CL protease,Mpro)和冠状病毒木瓜样蛋白酶(papain-like protease,PLP)的晶体结构,对中药化学成分数据库(traditional Chinese medicine database,TCMD 2009)进行基于分子对接的虚拟筛选,选择打分值排名前100的成分作为潜在抗新型冠状病毒活性的中药成分,并对成分来源中药进行频数统计,挑选命中成分数目较多的中药作为潜在抗新型冠状病毒活性的中药。研究结果显示,基于Mpro靶点筛选获得潜在抗新型冠状病毒活性成分12322个,代表性活性成分有紫菀五肽A、川芎嗪、丹酚酸B等,筛选得到的中药为生姜、紫菀、三七、川芎、丹参、干姜、瞿麦、大黄、肉苁蓉等;基于PLP靶点筛选获得潜在抗新型冠状病毒活性成分11294个,代表性活性成分有姜酮酚、白果醇、阿魏酸等,筛选得到的中药为党参、羌活、生姜、白果、川芎、瓜蒌、白芍、补骨脂、苦参、三七、当归等。结合文献报道的基于分子对接筛选的血管紧张素转换酶2(angiotensin converting enzyme 2,ACE2)靶点抑制活性候选中药,该文在湖南省新型冠状病毒肺炎中医药诊疗方案的基础上,将Mpro抑制、PLP抑制和ACE2抑制候选中药针对该疫情的合理应用进行了探讨,分析结果表明,PLP抑制活性筛选获得的瓜蒌及ACE2筛选获得的贝母包含于桑贝止嗽散、小陷胸汤之中,有清化热痰之功效,主治痰热壅肺所致的痰多咳喘或燥痰犯肺、干咳少痰、咯痰不爽,可应用于病毒作用初期外邪闭肺所致的咳嗽微喘等证型;Mpro抑制活性筛选获得的中药大黄佐以PLP筛选获得的瓜蒌,清泻腑气,二者包含于麻杏石甘汤、宣白承气汤中,可应用于疾病重症期邪热壅肺、疫毒闭肺以致腑实里结等证型;ACE2抑制活性筛选获得的中药桑叶、金银花及连翘包含于桑菊饮和银翘散中,轻清宣透,可应用于疾病初期温邪犯肺及咳嗽微喘等证型。研究结果旨在为进一步开展中医药应对新型冠状病毒疫情的治疗方案提供参考。

Study on screening potential traditional Chinese medicines against 2019-nCoV based on Mpro and PLP

With the rapid outbreak of COVID-19,traditional Chinese medicine(TCM)has been playing an active role against the epidemic.However,the screening of TCM is limited by the development cycle and laboratory conditions,which greatly limits the screening speed.This study established optimization docking models and virtual screening to discovery potential active herbs for the prevention and treatment of the novel coronavirus based on molecular docking technology.The crystal structures of 3 CL protease(Mpro)and papain-like protease(PLP)were obtained from PDB database and homologous modeling respectively,and were used to conduct virtual screening of TCMD 2009 database by CDOCKER program.The ingredients scored in the top 100 were selected respectively,and the candidate herbs were ranked by the numbers of hit molecules.Based on Mpro inhibitors screening,12322 potential active components were obtained,and the representative active components included aster pentapeptide A,ligustrazine,salvianolic acid B,etc.,and Zingiberis Rhizoma Recens,Asteris Radix et Rhizoma,Notoginseng Radix et Rhizoma,Chuanxiong Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma,Zingiberis Rhizoma,Dianthi Herba,Rhei Radix et Rhizoma,Cistanches Herba were obtained.While 11294 potential active ingredients were obtained by PLP inhibitor screening,representative active ingredients included gingerketophenol,ginkgol alcohol,ferulic acid,etc.,and Codonopsis Radix,Notopterygii Rhizoma et Radix,Zingiberis Rhizoma Recens,Ginkgo Semen,Chuanxiong Rhizoma,Trichosanthis Fructus,Paeoniae Radix Alba,Psoraleae Fructus,Sophorae Flavescentis Radix,Notoginseng Radix et Rhizoma,Angelicae Sinensis Radix were chosen.By combining the diagnosis and treatment scheme of Hunan province′s and angiotensin converting enzyme 2(ACE2)inhibitors screening from literature,present study also discussed the rational application of candidate herbs to this epidemic situation.Trichosanthis Fructus obtained by PLP inhibitors screening and Fritillaria verticillata obtained by ACE2 inhibitors screening were parts of the Sangbei Zhisou Powder and Xiaoxianxiong Decoction,which might be applicable to the syndromes of cough and dyspnea.Rhei Radix et Rhizoma screened by Mpro and Trichosanthis Fructus screened by PLP were contained in Maxing Shigan Decoction and Xuanbai Chengqi Decoction,and could be applied to the syndromes of epidemic virus blocking lung.Mori Folium,Lonicerae Japonicae Flos and Forsythiae Fructus obtained by ACE2 inhibitors screening were included in the Sangju Decoction and Yinqiaosan,which might be applicable to the syndromes of warm pathogen attacking lung and cough and dyspnea.The results of this study are intended to provide a reference for the further development of traditional Chinese medicine to deal with the new epidemic.