CAJ | 학술논문

目的利用假病毒技术,研究来源于马尾树树皮的化合物蜡果杨梅酸B抑制HIV-1进入的活性及作用机制.方法利用pHXB2和pVSV-G两个病毒包膜蛋白质粒,与pNL4-3.Luc.R-E-共转染,构建HIV-1 Env假病毒和VSV-G假病毒,检测化合物抑制假病毒感染的活性.采用针对包膜蛋白亚基gp41的ELISA方法,结合分子对接,研究活性化合物抗HIV-1的作用机制.结果从马尾树树皮中来源的2个三萜类单体化合物蜡果杨梅酸B(myriceric acid B)和蜡果杨梅酸C(myriceric acid C)中,仅蜡果杨梅酸B能特异性地抑制HIV-1 Env假病毒感染,其半数抑制浓度(IC_(50))值为(8.3±0.2) mg·L~(-1).此外,蜡果杨梅酸B在C-28位羧基的酯化产物蜡果杨梅酸B甲酯(myriceric acid B methyl ester)没有抑制HIV-1进入的活性.蜡果杨梅酸B的进入抑制活性与其抑制gp41六螺旋束结构形成的机制有关.分子对接表明,蜡果杨梅酸B能靶向gp41上N-螺旋三聚体的靶穴位置.结论蜡果杨梅酸B是作用于gp41的HIV-1进入抑制剂,其分子结构中C-28位的羧基及C-3位羟基与抑制活性密切相关.蜡果杨梅酸B可作为先导化合物来研发新的HIV进入抑制剂类抗艾滋病药物.
목적 이용가병독기술,연구래원우마미수수피적화합물사과양매산B억제HIV-1진입적활성급작용궤제.방법 이용pHXB2화pVSV-G량개병독포막단백질립,여pNL4-3.Luc.R-E-공전염,구건HIV-1 Env가병독화VSV-G가병독,검측화합물억제가병독감염적활성.채용침대포막단백아기gp41적ELISA방법,결합분자대접,연구활성화합물항HIV-1적작용궤제.결과 종마미수수피중래원적2개삼첩류단체화합물사과양매산B(myriceric acid B)화사과양매산C(myriceric acid C)중,부사과양매산B능특이성지억제HIV-1 Env가병독감염,기반수억제농도(IC_(50))치위(8.3±0.2) mg·L~(-1).차외,사과양매산B재C-28위최기적지화산물사과양매산B갑지(myriceric acid B methyl ester)몰유억제HIV-1진입적활성.사과양매산B적진입억제활성여기억제gp41륙라선속결구형성적궤제유관.분자대접표명,사과양매산B능파향gp41상N-라선삼취체적파혈위치.결론 사과양매산B시작용우gp41적HIV-1진입억제제,기분자결구중C-28위적최기급C-3위간기여억제활성밀절상관.사과양매산B가작위선도화합물래연발신적HIV진입억제제류항애자병약물.
Aim To investigate the HIV-1 entry inhibitory activities of myriceric acid B and C isolated from Rhoiptelea chiliantha Diels et Hand-Mazz and their mechanism of action.Method The plasmids encoding envelope proteins of HIV-1 (pHXB2) and VSV (pVSV-G) were cotransfected 293T cells with pNL4-3.Luc.R-E- to produce HIV-1 Env pseudovirus and VSV-G pseudovirus,respectively,which were used for testing the antiviral activities of these compounds.ELISA and molecular docking were used to study the mechanism of action of the active compounds.Results Myriceric acid B could significantly inhibit the infection of HIV-1 Env pseudovirus with an IC_(50) of(8.3±0.2)mg·L~(-1).The carbonoxyl group at C-28 position and the hydroxyl group at the C-3 position of myriceric acid B are important for its anti-HIV-1 activity.Like other HIV-1 entry inhibitors targeting gp41 (eg,ADS-J1 and NB-64), myriceric acid B could also block the gp41 six-helix bundle formation.Molecular docking analysis suggests that myriceric acid B may bind to the hydrophobic cavity of the gp41 N-trimeric coiled coil.Conclusion Myriceric acid B is a potent HIV-1 entry inhibitor targeting gp41 and can serve as a lead compound for developing novel anti-HIV-1 drug.