“天然植物抗菌液”(PAMs)潜在抗SARS-CoV-2活性成分及机制

Preliminary explorations of potential active components and mechanisms of“natural plant antimicrobial solution”(PAMs)against SARS-CoV-2

为探寻云南普洱丹州制药股份有限公司出品的“天然植物抗菌液”(PAMs)潜在抗新型冠状病毒(SARS-CoV-2)的可能活性成分及作用机制,应用TCMSP、BATMAN和YaTCM数据库查找PAMs中植物药材来源的化学成分和作用靶点;通过GeneCards数据库获取冠状病毒靶点;Cytoscape作图构建“药材-成分-靶点-疾病”网络和潜在的靶点PPI网络;GO和KEGG富集分析预测PAMs的作用机制,并将PAMs主要活性成分与新型冠状病毒的关键蛋白进行分子模拟与对接。最后,以M13噬菌体为病毒模型,测定PAMs对噬菌体病毒的抑制作用。共得到166种PAMs中的药材活性成分、1007个药物靶点,调控新型冠状病毒的潜在作用靶点73个。构建的药材-成分-靶点-疾病网络中,靶点度数大于10的有PTGS2、PTGS1、PPARG、DPP4、CALM1、CASP3、RELA、BAX等。GO富集分析共得到2109个条目,主要包括细胞因子、蛋白磷酸化、免疫调节等;KEGG共富集到144条信号通路,排名前5的依次为人巨细胞病毒感染、TNF、卡波西氏肉瘤相关疱疹病毒感染、甲型流感以及乙型肝炎信号通路。分子对接结果显示,木犀草素、槲皮素、黄芩素与SARS-CoV-2的3CL水解酶(Mpro)、S刺突糖蛋白均呈现出强的结合能力;黄芩素、杨梅素、槲皮素与血管受体紧张素转化酶(ACE2)的结合能力强。PAMs对M13噬菌体的抑制测定显示,20%的PAMs对其抑制作用达到99%以上。PAMs可能通过多成分、多靶点、多通路作用于SARS-CoV-2,并且红花、紫草的主要活性成分槲皮素、黄芩素以及木犀草素与Mpro、ACE2以及S刺突糖蛋白均有较低的结合自由能,揭示其可能直接对COVID-19产生辅助治疗作用。

In order to explore the possible active components and mechanisms of“natural plant antimicrobial solutions”(PAMs)produced by Yunnan Pu’er Danzhou Pharmaceutical Co.,Ltd.for SARS-CoV-2,the chemical components coming from the medicinal plants of PAMs and their drug targets were searched using TCMSP,BATMAN and YaTCM databases.The disease targets for coronavirus were obtained through the GeneCards database.Cytoscape mapping was applied to construct the“medicinal plant-component-target-disease”network and potential targets in PPI network.GO and KEGG enrichment analysis were utilized to predict the possible mechanism of PAMs.The main active components of PAMs were molecularly docked with the key proteins of SARS-CoV-2.Finally,M13 bacteriophages were used as the virus model for determination of the inhibitory effect of PAMs on virus.A total of 166 kinds of active components of PAMs,1007 kinds of drug targets,and 73 kinds of drug-coronavirus interaction targets were obtained.PTGS2,PTGS1,PPARG,DPP4,CALM1,CASP3,RELA and BAX were found to have target degrees greater than 10 in the medicinal plant-component-target-disease network.A total of 2109 entries were obtained by GO enrichment analysis,which were mainly involved in the regulations of cytokines,protein phosphorylation,and immunity.KEGG was enriched to 144 signaling pathways,and the top five were human cytomegalovirus infection,TNF,Kaposi’s sarcoma-associated herpesvirus infection,influenza A,and hepatitis B.The results of molecular docking showed that luteolin,quercetin and baicalein had strong binding ability to 3CL hydrolase(Mpro)and spike glycoprotein of SARS-CoV-2 while baicalein,myricetin,and quercetin had strong binding ability to angiotensin converting enzyme(ACE2).At concentration of 20%,PAMs could inhibit the growth of M13 to over 99%.In conclusion,PAMs might act on SARS-CoV-2 through multiple components,targets and pathways.The active components of quercetin,baicalein,and luteolin of PAMs mainly came from safflower and comfrey had lower binding free energy for Mpro,ACE2 and spike glycoprotein,indicating its possible adjuvant and therapeutic effects on COVID-19.