藏药材西北沼委陵菜抗HIV-1病毒物质基础及其作用机制研究

Looking for new anti-HIV-1 drugs for long-term use is an important and effective way to prevent and control of HIV spread in the further. Tibetan medicine has its advantages in multi-target effect, improving immunity, low price and controllable toxicity. Previously, Comarum salesovianum, a plant of Tibetan medicine, has been found clear anti-HIV effect by our team, specially, the n-butanol extract has the best anti-HIV-1 activity. The futher study found that the effective components of anti HIV-1 activity can not only inhibit integrase activity, but aslo can bind with auxiliary protein Vif in vitro to antagonize Vif degradation host protein A3G, suggesting that the drug may have new targets. In this proposal, we intend to figure out the effective components of Northwest marsh cinquefoil using natural organic chemistry strategy, which is expected to find a new target of drug. Meanwhile, chemical composition will be studied by activity-oriented and bioassay guide means and anti-HIV-1 activity material basis from Northwest marsh cinquefoil will be found. Its mechanism of anti-HIV-1 and structure-activity relationship will be illustrated. This study will clarify the new potential targets of drugs. All results of project will provide the material and theoretical basis for the development of innovative drugs with a clear target of anti HIV-1.

寻找可长期用于艾滋病感染者的抗HIV-1病毒新药是进一步增强防控艾滋病毒扩散的有效途径。天然藏药同时具有多靶点起效、增进免疫、价格低廉和毒性可控的优势。申请人团队前期发现藏医药用植物西北沼委陵菜具有明确的抗艾滋病毒功效，其正丁醇萃取部位具有抗HIV-1病毒活性。研究发现抗HIV-1病毒活性的有效成分不仅可以抑制整合酶的活性，还能与HIV-1的辅助蛋白Vif体外结合，拮抗Vif降解宿主A3G蛋白，提示药物可能具有新的作用靶点。本课题拟利用天然有机化学方法对其有效组分进行深入研究，有望发现药物可能作用的新靶点。同时开展以活性为导向的化学成分研究，通过活性追踪的方法，从西北沼委陵菜抗HIV-1病毒活性部位中发现抗HIV-1药效物质基础，明确其作用机制及构效关系，阐明药物的多靶点特性和可能的新靶点。为开发有明确靶点的抗HIV-1病毒的创新药物提供物质基础及理论基础。

项目从具有明确抗艾滋病毒功效的藏药材西北沼委陵菜正丁醇萃取部位中鉴定了62个化学成分，通过活性筛选，发现熊果酸、硬脂醇甘草亭酸酯、白桦脂醛、迷迭香酸、Khookerianic acid B是西北沼委陵菜中主要活性成分。计算机模拟实验表明，其中三萜类化合物与三种病毒蛋白活性结构域Intergase protein-4DMN、VIF-2ma9、VPU-1pi8均有很好的结合能力，并与VIF-2ma9蛋白的结合能力最强。假病毒单一生活周期感染实验，发现硬脂醇甘草亭酸酯可能与干扰病毒进入相关， Khookerianic acid B（phenolic lipid）可能与抑制逆转录和病毒进入相关。采用NanoBiT活细胞实时蛋白相互作用检测技术建立HIV-1 Vpu蛋白与宿主细胞BST-2蛋白的相互作用检测模型，发现迷迭香酸对Vpu-BST-2相互作用具有一定的抑制作用，提示抑制Vpu-BST-2互作可能是迷迭香酸抗病毒作用的机制之一。