结构新颖的萘醌色素和单萜苯酚类化合物的发现及抗HCV活性评价

The hepatitis C virus (HCV) infects about 170 million people worldwide. Its chronic infection is the cause of liver cirrhosis and hepatocellular carcinoma, and approximately 80% of infected individuals become chronically infected. Combination therapy of interferon-α and ribavirin formulation is the recommended treatment for HCV-infection. This combination therapy is limited by its only partial efficacy, and provokes severe side-effects. .As part of a search for anti-HCV agents of natural origin, a phytochemical and bioactive investigation of Arnebia euchroma was carried out, and naphthoquinones and monoterpene phenols were found to exhibit potent anti-HCV activity. In order to search for more bioactive natural products, we will initiate chemical and bioactive studies on naphthoquinones and monoterpene phenols from two Boraginaceae species (A. euchroma and Onosma paniculatum). Our future works will include: (1) The isolation and identification of novel naphthoquinones and monoterpene phenols from A. euchroma and O. paniculatum. (2) The evaluation of anti-HCV effects of these compounds on the basis of 2a chimeric strains (J6/JFH1), Huh 7.5.1 HCV cell culture system in vitro, with interferon and ribavirin as positive control. The toxicities of the compounds were assayed by a modified MTT method, the anti-HCV activity was assessed by detecting viral load with fluorescence quantitative PCR method and indirect immunofluorescence method, and the synergy, additivity or antagonistic interactions of effective compounds were determined by Isobologram analysis. (3) The primary investigation on the possible mechanism of anti-HCV activities of the bioactive compounds. .Taken together, these results will evaluate the anti-HCV activities of naphthoquinones and monoterpene phenols by pharmaceutical and toxicological research work, primarily ascertain their potential value in use.

丙型肝炎为一严重危害人类健康的传染病，感染着全球约1.7亿人口，约80%感染者可发展成慢性肝炎，甚至肝硬化和肝癌。目前临床主要应用干扰素和利巴韦林联合治疗丙型肝炎，但有接近 50％的患者对治疗不敏感或无反应，并伴有明显的副作用。.我们在前期研究中发现具有显著抗HCV活性的萘醌色素和单萜苯酚类化合物，项目组将在此基础上继续对新疆紫草和滇紫草的抗HCV活性部位进行研究，以发现结构新颖的单萜苯酚和萘醌色素类化合物。通过2a型嵌合病毒（J6/JFH1）感染Huh 7.5.1细胞的HCV体外细胞培养体系，对分离得到的化合物进行抗HCV活性筛选，分析两种活性化合物配伍的抗病毒作用。同时，对有明显抗HCV作用的化合物的作用靶点进行初步研究，探索其可能的作用机理，评价细胞毒性，为发现靶点明确、高效低毒的抗HCV活性化合物，为进一步开发新的天然来源的抗HCV药物奠定基础。

对滇紫草和新疆紫草的化学成分进行了研究。从滇紫草中共分离鉴定了36个化合物，10个为新化合物，结构类型涉及单萜苯酚、苯醌、苯并氢醌、紫草二聚体、丹参酮类及二聚体等。其中化合物phenshikonofurans A和 B (1-2)为结构新颖的萘醌类衍生物，其萘环上除了11位和14位连接1个1,2,4-取代的苯环外，12位碳还被α,β-二取代的呋喃环所取代。Naphthofurans A-C (15-17)和3-methoxyl-anhydroalkannin (18)是萘醌类衍生物，其中化合物15-17的C-5位和C-12位通过醚键形成一个骈合于萘醌苯环上的呋喃环，属于二氢苯并呋喃醌类化合物。Macrorhizonone (19)是首次从滇紫草中分离得到的苯并氢醌类新化合物，其2、3位被一个十元大环的单萜取代。Shikometabolin F (20)，shikometabolin G (21)和shikometabolin A (22)是以二聚形式存在的新的萘醌类衍生物。此外，从新疆紫草中分离得到了15个已知的紫草萘醌色素和单萜苯酚类成分。. 通过抗HCV活性筛选，发现化合物deoxyshikonin、shikonin和tanshinone II A具有显著的抗HCV活性，SI值分别为125.88、18.43和74.80。发现6个萘醌类化合物和2个菲琨类丹参酮化合物具有较为显著的抗炎活性，对NO抑制的IC50 在0.4–4.7 µg/ml之间，高于阳性对照L-NMMA；发现丹参酮II B具有一定的抗HIV活性；发现新化合物naphthofuranin C和macrorhizonone具有显著的抗甲型H1N1流感病毒活性，IC50 = 6.3+/-2.5, 8.3+/-3.1 µg/ml，CC50 = 40+/-3.5和24+/-2.1 µg/m。对活性较好的萘醌类化合物shikonofuran E的抗炎作用机理进行了研究，发现该化合物通过抑制MAPKs和NF-κB两条信号通路下调炎症因子TNFα、IL-1β和IL-6以及相关蛋白COX2和iNOS的表达，发挥抗炎作用。. 已发表标注基金资助的学术论文12篇，其中SCI收录11篇，发表中文核心期刊1篇；获国家发明专利2项。已培养博士研究生1名，硕士研究生2名。