HIV-1双靶点抑制剂QCA-DAPYs杂合物的设计、合成及抗HIV活性研究

Human immunodeficiency virus type-1 reverse transcriptase (HIV-1 RT) and integrase (IN) are ones of the virally encoded enzymes required for replication and therefore represent rational targets for the treatment of HIV-1 infection. In our previous work, we have obtained a series of new quinolone-3-carboxylic acids (QCAs) as HIV-1 IN inhibitors and diarylpyrimidines (DAPYs) as RT inhibitors. Based on Morphy's multi-target drug strategy, in this project, we planed to design of a novel series of QCA-DAPY hybrids as HIV-1 dual inhibitors by merging the scaffolds of these two types of inhibitors and docking study of binding mode of these hybrids with RT and IN. The designed hybrids that could adopt right orientation in the binding site of both RT and IN will be synthesized and tested for their HIV-1 IN and RT inhibitory assay, as well as the anti-HIV activity. 3D-QSAR model will also be developed. 1 to 3 target compounds with strong antiviral potency against both HIV-1 wild-type and mutant virus will be further screened for their metabolic stability. It is possible to obtain 1 to 2 new anti-HIV candidates with high anti-HIV potency and metabolic stability in this project. The expected research results would provide a theoretical basis for the development of our own new anti-HIV drugs.

HIV-1整合酶（IN）和逆转录酶（RT），是HIV生命复制周期中的必需关键酶，成为设计抗HIV药物的重要靶点。本项目基于Morphy“一药多靶点”设计新策略，拟将前期研究获得的QCA类IN抑制剂和DAPY类RT抑制剂进行重叠整合，获得系列QCA-DAPYs杂合物。并将这些杂合物分别与IN、RT进行对接研究，筛选能够同时在两种靶酶结合口袋均可形成正确空间取向的化合物，以此设计HIV-1双靶点抑制剂QCA-DAPYs杂合物。通过化学手段合成目标物，并对目标物进行IN、RT酶水平及细胞水平的抗HIV活性筛选，建立3D-QSAR模型，筛选1-3个高活性强抗耐药性化合物进行离体生物代谢稳定性研究。本项目力争从中发现1-2个高活性、强抗耐药性、且代谢稳定的抗HIV新药候选物，为研发具有我国自主知识产权的抗HIV药物提供理论基础和技术支持。

HIV感染导致的艾滋病是人类历史上危害最深广、灾难最严重的传染病之一。现有19种RT抑制剂和2种IN抑制剂被FDA批准用于抗AIDS临床治疗，但这些单靶点抗HIV药物在使用后快速产生的耐药性严重限制了其临床应用。随着HIV耐药性日益严重，寻找新型高效抗HIV药物已迫在眉睫。.基于Morphy“一药多靶点”设计策略，本申请将前期研究获得的QCA类HIV-1 IN抑制剂和DAPY类RT抑制剂进行重叠整合，设计、合成了一类新型HIV-1抑制剂QCA-DAPY类杂合物。共完成36个HIV-1 RT/IN双靶点抑制剂QCA-DAPYs杂合物的分子设计、合成及其抗HIV活性筛选（包括：HIV-1野生株、双突变耐药株）,发现目标化合物12ag同时对HIV-1野生株及双变异耐药株均具有强抑制活性（EC50 （HIV-1 IIIB）: 0.0096 μM, EC50 （K103N/181C）: 0.0096 μM），但选择指数SI低于参考药物伊曲韦林，仍需进一步进行结构优化，以期获得值得进一步进行代谢研究的候选药物。通过一系列酶活性抑制实验（RT polymerase、RNase H、IN、Time-of-addition assay）、EC50/IC50相关性、以及分子对接，对目标化合物的作用机制进行了深入的研究, 发现目标化合物12o同时对HIV-1逆转录酶和整合酶具有μM级别的抑制活性，为寻找极具挑战性的RT/IN双靶点抑制剂提供了苗头化合物。此外，对所合成的目标分子进行了构效关系归纳总结，建立了预测能力良好的3D-QSAR模型。这些研究结果为研发具有我国自主知识产权的抗HIV药物提供理论基础和技术支持。