鱼腥草素杂合黄酮类抗单纯疱疹病毒活性成分的优选和机制研究

Herpes simplex virus (HSV) infection is a highly contagious disease of human beings, and drug resistance and low bioavailability are common problems in the treatment of this disease. It is very important to discover anti-HSV drugs from the resource-rich traditional Chinese medicine (TCM). In the previous research of our team on the anti-HSV active constituents of Houttuynia cordata, a kind of novel skeleton flavonoids, named “houttuynoids” was firstly obtained, which showed anti-HSV activity. The preliminary mechanism indicated that houttuynoid A specially targeted at HSV fusion process, which was different from that of first-line anti-HSV drugs. Notably, an abundance of unidentified houttuynoids were detected by HPLC-MS. Based on the above research, the target isolation of houttuynoids from Houttuynia cordata will be carried out. Meanwhile, metabolic regulation of Houttuynia cordata callus and microbial transformation will be applied to enrich chemical diversity of houttuynoids. Plaque reduction assay will be carried out to evaluate the anti-HSV activity of houttuynoids, followed by the establishment of the structure–anti-HSV activity relationship (SAR) of houttuynoids. Considering SAR and the druggability, the anti-HSV candidates will be optimal selected and enriched. Then, the mice lip HSV infection model will be performed to evaluate anti-HSV effects of the candidates and explore their anti-HSV mechanism. In conclusion, this project intends to optimally select anti-HSV candidate drugs with explicit mechanism, and to lay the foundation for the development of natural anti-HSV drugs.

单纯疱疹病毒（HSV）感染是人类高发的传染病，临床用药普遍存在耐药性和生物利用度低等问题，从资源丰富的中药中发掘抗HSV药源分子具有重要意义。项目前期从鱼腥草中发现了一类全新骨架黄酮类成分，命名为“鱼腥草素杂合黄酮（houttuynoids）”，具有抗HSV活性；对houttuynoid A初步的机制研究揭示其作用于病毒侵染早期，有别于目前一线用药。液质分析发现鱼腥草中还存在大量未被鉴定的同类成分，值得深入研究。申请人拟在此基础上，采用液质靶向分离鱼腥草中富含的同类成分，辅以鱼腥草愈伤组织代谢调控和微生物转化等方法获得结构多样的该类成分；采用空斑减数法评价其抗HSV活性，探讨构效关系，结合成药性考察优选活性最强、成药性最佳的候选分子；开展候选分子定向富集制备和小鼠HSV口唇疱疹感染药效学评价，揭示其作用机制，以期获得药效更强、机制明晰的抗HSV候选药物，为天然来源抗HSV新药研制奠定基础。

项目前期从鱼腥草中发现了一类全新骨架黄酮类成分，命名为“鱼腥草素杂合黄酮（houttuynoids）”，具有抗HSV活性。液质分析发现鱼腥草中还存在大量未被鉴定的同类成分。基于此，项目采用前期建立的鱼腥草素杂合黄酮类成分的质谱裂解规律和快速检识策略从鱼腥草提取物中靶向分离获得了17个该类成分（10个新化合物）；采用空斑形成法评价了鱼腥草素杂合黄酮类成分的抗HSV-1活性。活性评价结果显示，所测试的化合物均具有一定程度的抗HSV活性，其中houttuynoid M、houttuynoid L和houttuynoid A的抗HSV-1活性较强。根据活性结果、药源等因素，优选出houttuynoid A是鱼腥草素杂合黄酮抗HSV的候选分子。同时定向制备获得了克级的houttuynoid A样品。接着，项目对houttuynoid A等克级化合物进行了预防HSV-1感染的体内外活性进行药效学评价，结果显示houttuynoid A预保护给药主要通过影响细胞融合发挥作用，但其不能通过预保护给药途径来增强固有免疫以拮抗HSV-1诱导的脑炎模型。进一步地，项目还扩展了研究内容，采用HPLC-MRM-MS方法分析了不同生长周期的鱼腥草不同药用部位（根、茎、叶）的鱼腥草素杂合黄酮类等成分的含量动态变化情况。结果显示不同药用部位在不同生长时期的成分含量相差较大，鱼腥草素杂合黄酮主要分布在鱼腥草叶中，其总含量随着普通黄酮氧苷类成分含量的增加而降低。.此外，部分研究成果已发表SCI论文1篇，已获受理国内发明专利1项，正在培养硕士研究生3名。