大蓟内生真菌Coniothyrium sp. DJ-1抗SARS-CoV-2 M^(pro)活性的化学成分研究  

Chemical constituents with potential anti-SARS-CoV-2 M^(pro) activity from endophytic fungus Coniothyrium sp.DJ-1 of Cirsium japonicum

作　　者：曾洁醇 杨润 黄金 李艳平 高路[1] 汪伟光 ZENG Jie-chun;YANG Run;HUANG Jin;LI Yan-ping;GAO Lu;WANG Wei-guang(Key Laboratory of Natural Products Synthetic Biology of Ethnic Medicinal Endophytes,State Ethnic Affairs Commission,School of Ethnic Medicine,Yunnan Minzu University,Kunming 650031,China;College of Traditional of Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南民族大学国家民委民族药内生菌天然产物合成生物学重点实验室,民族医药学院,云南昆明650031 [2]云南中医药大学中药学院,云南昆明650500

出　　处：《中草药》2023年第10期3071-3079,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金项目(31960095);国家自然科学基金项目(82160670);云南省应用基础项目(202101AS070022,202201AT070228);云南省万人计划-青年拔尖人才(W.-G.Wang)。

摘　　要：目的研究大蓟Cirsium japonicum内生真菌Coniothyriumsp.DJ-1中抑制新型冠状病毒主蛋白酶(severea cute respiratory syndrome coronavirus 2 main protease,SARS-CoV-2 M^(pro))活性的化学成分。方法运用新型冠状病毒M^(pro)/3CLpro抑制剂筛选模型从30株菌株中筛选具有抑制活性的菌株和化学成分。采用正相硅胶、反相RP-18柱色谱和HPLC等技术对抑制率高的菌株的次生代谢产物进行分离纯化,通过NMR、质谱等谱学技术解析化合物结构,利用荧光共振能量转移法测定化合物对SARS-CoV-2 M^(pro)的抑制活性。结果在30株菌株中发现抑制活性良好的菌株Coniothyrium sp.DJ-1,并从该菌的大米发酵提取物中分离到17个化合物,分别鉴定为德加隆内酯(1)、5’-表细链格孢烯(2)、细链格孢烯(3)、4’-表细链格孢烯(4)、3-表-二氢细链格孢烯A(5)、瓶霉酚(6)、1-脱氧红藻内酯(7)、6-羟基-8-甲氧基-3a-甲基-3a,9b-二氢-3H-呋喃并[3,2-c]异色烯-2,5-二酮(8)、黄色篮状菌酮(9)、链格孢毒素II(10)、二氢链格孢苝醇(11)、2-(2’-羟基丙基)-5-甲基-7-羟基色酮(12)、7-羟基-2-羟甲基-5-甲基-4H-色烯-4-酮(13)、三环链格孢霉烯6b(14)、三环链格孢霉烯D(15)、苯乙醇(16)和对羟基苯甲醛(17)。化合物5、9、11对SARS-CoV-2 M^(pro)的半数抑制浓度(median inhibition concentration,IC50)分别为42.77、25.85和24.26μmol/L。结论化合物1~15和17均为首次从Coniothyriumsp.属中分离得到,化合物5、9、11对SARS-CoV-2 M^(pro)有较好的抑制作用。通过筛选活性菌株的策略,快速从真菌天然产物中挖掘抗新冠病毒活性的化合物,为抗新冠病毒药物的研发提供了参考。Objective To study the chemical constituents with severe acute respiratory syndrome coronavirus 2 main protease(SARS-CoV-2 M^(pro))inhibitory activity from Coniothyrium sp.DJ-1,an endophytic fungus isolated from Cirsium japonicum.Methods The strains and chemical constituents both with inhibitory activity were screened from thirty endophytic fungi by using 2019-nCoV M^(pro)/3CLpro inhibitor screening model.The secondary metabolites of the strain with high inhibition rate were separated and purified by normal phase silica gel column chromatography,reversed-phase RP-18 column chromatography,and HPLC.Their structures were identified by NMR,MS,and optical rotation data.And the compounds’anti-SARS-CoV-2 M^(pro) activities were determined by fluorescence resonance energy transfer method.Results The Coniothyrium sp.DJ-1 strain with good inhibitory activity was screened from thirty endophytic fungi.Seventeen compounds were isolated from the rice fermentation extract of Coniothyrium sp.DJ-1,and identified as djalonensone(1),5'-epialtenuene(2),altenuene(3),4'-epialtenuene(4),3-epi-dihydroaltenuene A(5),phialophoriol(6),1-deoxyrubralactone(7),6-hydroxy-8-methoxy-3a-methyl-3a,9b-dihydro-3Hfuro[3,2-c]isochromene-2,5-dione(8),talaroflavone(9),altertoxin II(10),dihydroalterperylenol(11),2-(2'-hydroxypropyl)-5-methyl-7-hydroxychromone(12),7-hydroxy-2-hydroxymethyl-5-methyl-4H-chromen-4-one(13),tricycloalternarene 6b(14),tricycloalternarene D(15),2-phenylethan-1-ol(16)and 4-hydroxybenzaldehyde(17).Compounds 5,9 and 11 exhibited potent inhibitory effects on SARS-CoV-2 M^(pro) with IC50 values of 42.77,25.85 and 24.26μmol/L,respectively.Conclusion Compounds 1—15 and 17 have been isolated from the extracts of Coniothyrium sp.genus for the first time.Compounds 5,9 and 11 exhibit potent inhibitory effects on SARS-CoV-2 M^(pro).The research concludes that the method of screening active strains is helpful for us to quickly excavate active compounds from natural fungal products,moreover provides a basis for the research and developme

关 键 词：大蓟 盾壳霉属 内生真菌 次生代谢产物 抗病毒活性 新型冠状病毒主蛋白酶 3-表-二氢细链格孢烯A 黄色篮状菌酮 二氢链格孢苝醇 

分 类 号：R284.1[医药卫生—中药学]