摘要
本文研究了N4-(杂)芳基磺酰基喹喔啉酮类化合物6-氯-3-甲基-4-(2-甲氧羧基噻吩-3-磺酰基)-3,4-二氢喹啉-2-(1H)-酮(XU07011)抗HIV-1作用机制及特点。采用细胞水平模型评价该化合物抗HIV-1活性;通过在不同时间点加入化合物检测化合物失效时间,确定化合物抑制病毒复制的具体环节;酶联免疫吸附法和荧光法测定化合物对逆转录酶的DNA聚合酶活性和核糖核酸酶H(RNase H)催化活性的影响;应用非核苷类逆转录酶抑制剂(non-nucleoside reverse-transcriptase inhibitors,NNRTIs)耐药逆转录酶和重组病毒模型分别在酶和细胞水平研究化合物的作用特点。结果显示,XU07011可剂量依赖性地抑制HIV-1复制,IC50为(0.057±0.01)μmol·L?1,活性与奈韦拉平相当;该化合物作用于HIV-1复制的逆转录环节,抑制了逆转录酶的RNA依赖DNA聚合酶活性,IC50为(1.1±0.3)μmol·L?1,而对逆转录酶RNase H活性无显著影响;该化合物对9种NNRTI耐药病毒的耐药倍数为33～2000倍,其中对K103N耐药病毒活性优于奈韦拉平。本研究为新型HIV-1抑制剂的研发提供了物质基础。
This study is to investigate the mechanism and action characteristics of 6-chloro-3-methyl-4-（2- methyoxycarbonylthiophene-3-sulfonyl）-3, 4-dihydroquinoxa-lin-2-（1 H）-one （XU07011） against HIV- 1 replication. XU07011 anti-HIV activity was tested by using VSVG/HIV pseudotype viral system and confirmed by HIV-1 live viruses＇ infectious assay. Time of addition was used to test HIV-1 reverse transcription process. RNA- dependent DNA polymerase activity and RNase H activity were tested by using enzyme linked immunoabsorbent assay and fluorescence method. Wild type and nine NNRTIs-resistant reverse transcriptase enzymatic models and cell-based pharmacological models were used to evaluate XU07011 bio-characteristics. The results showed that XU07011 inhibited HIV-1 replication with IC5o of （0.057 ±0.01） μmol.L-1 which was comparable to nevirapine [IC5o： （0.046± 0.01） μmol-L-1. Mechanism study data indicated that XU07011 blocked HIV-1 reverse transcription process through acting on reverse transcriptase RNA-dependent DNA polymerase with IC 50 of （1.1 ± 0.3） pmol-L-1. The compound showed no effect on RNase H activity. XU07011 exhibited better activities comparing with nevirapine on K103N mutated NNRTIs-resistant HIV-1 strains. This study could provide a theoretical basis for novel anti-HIV reagents development.
出处
《药学学报》
CAS
CSCD
北大核心
2013年第6期860-865,共6页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81273561)
“十二五”重大专项资助项目(2013ZX10004601)
