通过高通量药物库筛选挖掘针对金黄色葡萄球菌的新型抗菌药物的研究  

Antimicrobials discovery against Staphylococcus aureus by high throughput screening of drug library

作　　者：佘鹏飞 杨艺钒 李琳惠 周林颖 伍勇 She Pengfei;Yang Yifan;Li Linhui;Zhou Linying;Wu Yong(Department of Laboratory Medicine,the Third Xiangya Hospital,Central South University,Changsha 410013,China;Department of Laboratory Medicine,the Affiliated Changsha Hospital of Xiangya School of Medicine(the First Hospital of Changsha),Central South University,Changsha 410005,China)

机构地区：[1]中南大学湘雅三医院检验科,长沙410013 [2]中南大学湘雅医学院附属长沙医院(长沙市第一医院),长沙410005

出　　处：《中华预防医学杂志》2023年第11期1855-1861,共7页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金 (82072350,82202591)。

摘　　要：通过高通量药物库筛选挖掘针对金黄色葡萄球菌(S.aureus,简称:金葡)的新型抗菌药物。本研究的类型为实验性研究。金葡临床菌株均于2017年1月至2019年1月分离自中南大学湘雅三医院呼吸科住院患者的痰液标本;通过摇菌培养浮游菌,检测美国食品药品监督管理局(FDA)认证药物库(包含1573种小分子)对金葡浮游菌增殖抑制作用;通过96孔板结合结晶紫染色,检测FDA认证药物库分子对金葡生物被膜形成的抑制作用;通过微量肉汤稀释实验,检测纳入小分子的最低抑菌浓度(MIC);最后,通过CCK-8实验检测小分子药物的细胞毒性。结果显示,通过将FDA认证药物库中的小分子浓度设置为100μmol/L,初步筛选出218种对金葡浮游菌具有生长抑制作用的小分子。这些小分子以抗感染药物为主,占118种。其次为抗肿瘤药物、抗炎/免疫类药物、神经系统药物、心血管系统药物、内分泌系统药物和代谢疾病药物,分别占40、19、12、9、8和3种,未分类药物占9种。排除抗感染药物后,针对具有金葡浮游菌生长抑制作用排名前十的小分子主要为抗肿瘤药物,其次为神经系统药物和未分类药物维生素K3,其抑制率均达99.65%~100%。针对具有金葡生物被膜抑制作用的排名前十小分子也以抗肿瘤药物占比最高,其次为神经系统药物和抗炎/免疫类药物,其抑制率为50.22%~92.95%。通过微量肉汤稀释实验检测了第二轮筛选得到的51个小分子的MIC。其中,主要包括抗肿瘤药物、心血管系统药物、内分泌系统药物、抗炎/免疫药物、代谢疾病药物、神经系统药物和其他未分类药物,分别占22、5、3、9、2、5和5种,其MIC分布分别为1.56~50μmol/L、6.25~25μmol/L、6.25~25μmol/L、0.2~50μmol/L、25~50μmol/L、1.56~50μmol/L和0.1~12.5μmol/L。综上,通过高通量药物库筛选出的小分子最低抑菌浓度值均为微摩尔级别,成药能力强且可改造性高。其高�To develop antimicrobials against Staphylococcus aureus by high throughput screening of drug library.The type of this study is experimental research.The clinical isolates of S.aureus were collected from the sputum samples of respiratory inpatient department of the Third Xiangya Hospital of Central South University.The anti-planktonic cells growth inhibition activity of FDA-approved drugs library(including 1573 molecules)was assessed by building a planktonic cells screening platform;The biofilm inhibitory effect of the FDA-approved drugs was detected by building a biofilm screening platform combined with crystal violet staining;Minimal inhibitory concentrations of the selected hits were determined by broth microdilution assay.Finally,the cytotoxicity of the selected hits was detected by CCK-8 assay.The results showed that 218 hits were exhibited effective growth inhibitory effects against S.aureus by setting the concentrations of the molecules in the FDA-approved library to 100μmol/L.These selected molecules are mainly anti-infective drugs,accounting for 118 hits;Followed by anti-cancer drugs,anti-inflammatory/-immune drugs,neurological drugs,cardiovascular drugs,endocrine drugs,and metabolic disease drugs,which accounts for 40,19,12,9,8,and 3 hits;Other unclassified drugs accounts for 9 hits.The top 10 hits exhibiting anti-planktonic cells activity against S.aureus were mainly including antitumor drugs,followed by neurological drugs and unclassified drugs like vitamin K3 with the inhibition rate of 99.65%-100%.Similarly,the top 10 hits showing biofilm inhibitory effects against S.aureus were also mainly including antitumor drugs,followed by neurological drugs and anti-inflammatory/-immune drugs with the inhibition rate of 50.22%-92.95%.The minimal inhibitory concentration(MIC)of the 51 hits by second round screening was determined by micro-dilution assay,which mainly include the antitumor drugs,cardiovascular drugs,endocrine drugs,anti-inflammatory/-immune drugs,metabolic disease drugs,neurological drugs and oth

关 键 词：金黄色葡萄球菌 抗菌药物 药敏实验 生物被膜 药物研发 

分 类 号：R446.5[医药卫生—诊断学]