多靶点药物设计是研发高效抗耐药HIV抑制剂的新策略；基于CuAAC点击化学的片段组装及原位筛选是具有广阔应用前景的药物先导物发现新技术。本课题为提高萘啶酮及N-羟胺类HIV RNase H-IN双靶点抑制苗头化合物的抗病毒活性及成药性、调节对靶点的特异性，根据RNase H与 IN三维结构、抑制剂药效团及抗耐药策略，运用多靶点药物设计策略及基于CuAAC点击化学高效合成筛选技术，对苗头化合物的金属螯合区及疏水作用区进行结构多样性的改造，构建以三唑为Linker的聚焦型RNase H-IN双靶点虚拟化合物库；通过多层级虚拟筛选确定目标化合物结构，优选取代炔或叠氮为优势片段实施化学合成并构建组合库，经原位抑酶活性筛选发现活性分子；继而进行扩大量合成、抗HIV活性及成药性评价发现抗HIV药物侯选物，并借助计算机技术探讨构效关系和结合模式。经几轮设计，为开发具有自主知识产权的抗HIV新药奠定基础。

Emergence of drug-resistant HIV-1 mutants and side effects impede the long-term usage of drugs, requiring a significant need for new strategies or protocols of drug discovery. Design of multitarget-directed ligands (MTDLs) has gained increasing acceptance; Meanwhile, fragment-based assembly using Huisgen Cu(I)-catalyzed alkyne-azide cycloaddition (CuAAC, a paradigm of click chemistry) combined with in situ screening has become a widely used strategy for high-throughput drug discovery and chemical space exploration of lead compounds. In order to improve drug resistance profile, druggability and lower cytotoxicity of reported hit molecules (hydroxynaphthyridinones and hydroxyisoquinoline-1,3-diones) with RNase H-IN dual inhibition, in this project, based on the two-metal binding pharmacophore model of RNase H-IN dual inhibitors, we start an analoging-based optimization program aimed at the exploration of the chemical diversity in metal binding group (MBG) and hydrophobic group (HG) by constructing 1,2,3-triazole linked molecular library using the CuAAC click reaction. By the hierarchical multiple-filter (binding affinity, drug-like profiles) virtual database searching strategy to predict the favorable molecules from a large collection of 1,4-disubstituted 1,2,3-triazoles, the top-ranking compounds will be selected and be dissected into a set of alkyne-containing MBGs and azide-containing HGs fragments. Initially, a library of 120-membered triazoles between 10 alkynes and 12 azides is rapidly constructed using CuAAC reaction and tested against RNase H and IN without purification (in situ screening). From the preliminary inhibition data, the most potent library members were individually resynthesized in large-scale for complete characterization and further assay of their anti-HIV activities in vitro and in vivo and their preliminary evaluation of drug-like properties. The binding modes of selected bioactive compounds and QSAR will be investigated by molecular simulations. Our approach thus lays the foundation for discovery of novel RNase H-IN dual inhibitors with improved drug resistance profile and favorable druggability.