牛奶外泌体负载利福霉素S衍生物的抗菌活性评价  

Evaluation of antimicrobial activity of milk exosomes loaded with rifamycin S derivative

作　　者：杨占群 李想[2] 刘成华 郑梦竹 樊士勇 董玉超 王子豪 林坚 杨光 陈龙 YANG Zhanqun;LI Xiang;LIU Chenghua;ZHENG Mengzhu;FAN Shiyong;DONG Yuchao;WANG Zihao;LIN Jian;YANG Guang;CHEN Long(Department of Pharmacy,Peking University Third Hospital,Beijing 100191,China;Second Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Acacemy of Military Medical Sciences,Beijing 100850,China;School of Pharmaceutical Sciences,Hainan University,Haikou 570228,China;College of Life Sciences,Anhui Medical University,Hefei 230032,China;Beijing Institute of Tropical Medicine,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]北京大学第三医院药学部,北京100191 [2]解放军总医院第二医学中心,北京100853 [3]军事医学研究院,北京100850 [4]海南大学药学院,海南海口570228 [5]安徽医科大学生命科学学院,安徽合肥230032 [6]首都医科大学附属北京友谊医院北京热带医学研究所,北京100050

出　　处：《中国药理学与毒理学杂志》2025年第3期208-215,共8页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的 设计合成3种利福霉素S衍生物,进行牛奶外泌体负载,并评价其体外抗菌活性。方法 合成利福霉素S衍生物,利用质谱和核磁进行化合物表征;使用稀释测定法测定3种利福霉素S衍生物对金黄色葡萄球菌(金葡菌)和铜绿假单胞菌的抑菌活性,并计算半抑制浓度(IC_(50));选择抗菌活性优异的衍生物,采用超声法将其负载到牛奶外泌体内,制备牛奶外泌体负载的利福霉素S衍生物,并采用稀释测定法测定其抗金葡菌活性;采用平板菌落计数法检测其对巨噬细胞内驻留金葡菌的抑菌作用。结果 成功合成3种利福霉素S衍生物,相较于母体化合物,利福霉素S衍生物对金葡菌(母体化合物的抗菌活性仅为3种衍生物抗菌活性的1/2~1/80)和铜绿假单胞菌(母体化合物的抗菌活性分别为利福霉素S衍生物1和3的1/14和1/9)的抑菌活性显著提高。成功实现牛奶外泌体对利福霉素S衍生物3的负载,负载率为10.9%。外泌体负载后的化合物对体外金葡菌和巨噬细胞内驻留的金葡菌的抗菌活性显著增强。结论 设计合成的利福霉素S衍生物具有更强抗菌活性,且外泌体负载后可增强其对胞外和胞内的抗菌活性。OBJECTIVE To design and synthesize rifamycin S derivatives and load them into milk exosomes to evaluate their in vitro antimicrobial activity.METHODS Rifamycin S derivatives were synthe-sized and characterized by mass spectrometry and NMR.Using the dilution assay method,the inhibitory activity of each rifamycin S derivatives molecule against Staphylococcus aureus and Pseudomonas aerugi-nosa was determined,and the IC50 was calculated.Derivatives molecules with excellent antimicrobial activity were selected and loaded into milk exosomes using the ultrasonication method,resulting in the preparation of milk exosome-loaded rifamycin S derivatives.The antimicrobial activity against Staphylo-coccus aureus was determined using the dilution assay method.The inhibitory effect of the exosome-loaded rifamycin S derivatives on Staphylococcus aureus residing within macrophages was detected using the plate colony counting method.RESULTS Three rifamycin S derivatives were successfully designed and synthesized,which demonstrated superior antimicrobial activity against Staphylococcus aureus(the parent compound′s antimicrobial activity is merely from 1/20 to 1/80 of that of the three rifamycin S derivatives)and Pseudomonas aeruginosa(the parent compound′s antimicrobial activity is only 1/14 and 1/9 of that of compound 1 and compound 3)compared to the parent compound.The loading of milk exosomes with the rifamycin S derivatives compound 3 was successfully achieved,with a loading efficiency of 10.9%.The antimicrobial activity of the compound after exosome loading was significantly enhanced against Staphylococcus aureus in vitro and against Staphylococcus aureus residing within macrophages(P<0.01).CONCLUSION The designed and synthesized derivatives of rifamycin S possess stronger anti-microbial activity,and their antibacterial efficacy against both extracellular and intracellular bacteria can be further enhanced after loading into exosomes.

关 键 词：利福霉素S衍生物 金黄色葡萄球菌 铜绿假单胞菌 外泌体 抗菌活性 

分 类 号：R962[医药卫生—药理学]