基于无定型纳米修饰技术的藏药波棱瓜子抗乙肝病毒木脂素活性成分发现

Herpetospermum caudigerum Wall. seed is a kind of common Ethnomedicine (EM) for the treatment of hepatitis, and has a long history and definite effect. However, the poor solubility of some components led to many originally valuable active ingredients abandoned and made the separated ingredients from TCM(ED) more and more ineffective. In addition, it was also the common key problem in phytochemistry field..The project aims to solve the international problem of HbeAg seroconversion rate and discover the poor soluble and potential highly active lignans from Herpetospermum caudigerum Wall. using the nano-amorphous (NN) key technology. Based on the preliminary screened abundant lignans, we will further separate the new lignans. Then the project will build the NN technical system on the basis of the structural parameters and physicochemical properties of lignans, and explain the NN’s modification rule in improving the poor solubility and low bioavailability of the potential active ingredients and get the key technology. The project will evaluate in vitro cell activity and in vivo anti-hepatitis B virus on the active ingredients before or after NN modified. The PK/PD experiment will be carried out at the same time to explore the antiviral active ingredients and structure-activity. On the basis of chemical structure, modification, and activity evaluation, the “structure-modification-activity ” rule of lignans will be obtained. The project has faced with the fact at the first time that the poor soluble as well potential highly active ingredients is abandoned, in addition to settle the common key problems by NN technology and promote the development of EM industry.

藏药波棱瓜子是藏医治疗肝病(赤巴病)的常用药物，历史悠久，疗效确切。由于成分难溶问题，使本来有价值的活性成分被抛弃，这亦是导致中药（民族药）越分越细越无效的重要原因之一，也是植物化学领域的共性问题。.本项目运用无定型纳米(NN，nano-amorphous)技术，瞄准HBeAg血清学转换难题，用于波棱瓜子中难溶、潜在高活性的木脂素发现。在项目组已获得大量木脂素成分基础上，进一步分离鉴定新型木脂素；基于成分结构参数及理化性质参数构建NN技术体系；阐释NN对改善难溶、低生物利用度活性成分的修饰规律，凝炼形成关键技术；开展NN改良前后成分的体外细胞和体内抗乙肝病毒活性评价；开展NN改良前后活性成分的PK/PD研究；在化学结构、改良修饰和效应的基础上，获得木脂素“结构-改良-效应”之间的规律。本项目首次正视难溶性、潜在高活性的化合物被摈弃事实，利用NN技术，解决行业共性关键问题，推动民族医药发展。

藏药波棱瓜子是藏医治疗肝病(赤巴病)的常用药物，历史悠久，疗效确切。由于成分难溶问题，使本来有价值的活性成分被抛弃，这亦是导致中药（民族药）越分越细越无效的重要原因之一，也是植物化学领域的共性问题。.本项目运用无定型纳米技术，瞄准HBeAg血清学转换难题，将制剂学手段前移，用于波棱瓜子中难溶、潜在高活性的木脂素发现，主要完成以下内容：一、采用高压制备色谱法进行木脂素成分的分离纯化，并通过UV、IR、NMR、MS等鉴定获得的单体成分，共分离制备了8个化合物，经鉴定分别是dehydrodiconiferyl alcohol、 herpetotriol I、 herpetolide A、 herpetrione、 herpetin、 herpetetrone、 herpetone、 herpetenone，其中herpetotriol I为新化合物。二、采用微型化介质研磨法和超声沉淀法制备木脂素成分纳米混悬剂，各木脂素成分制成纳米混悬剂后，粒径显著减小，体外溶出显著增加。三、以HSC-T6细胞和HepG2.2.15细胞为模型，对改良前后的木脂素体外抗肝纤及抗乙肝病毒活性进行了评价。结果显示，herpetrione (Her)、herpetone (HPT)、herpetolide A (HPA)、herpetetrone (HPTR) 对HSC-T6细胞增值均有显著的抑制作用，制成纳米混悬剂后其抑制作用显著增强； Her、HPA和HPTR均可显著抑制HepG 2.2.15细胞HBsAg、 HBeAg的分泌以及HBV-DNA的复制，但HPA纳米混悬剂作用显著强于HPA。而HPTR纳米混悬剂对HPTR抗乙肝作用改善不明显。药代动力学研究结果显示， HPA、HPTR制成纳米混悬剂后，Cmax分别增加了58.21%和56.88%，AUC0-24 分别增加了45.88%和85.90%。上述结果表明纳米混悬剂可明显改善波棱瓜子木脂素成分的口服生物利用度，提高抗肝纤活性及抗乙肝疗效。四、应用ACQ探针研究Her-NS体内吸收分布行为。结果表明Her-NS口服后并没有在体内迅速完全地溶出释放，而是能够在胃肠道内保留12~16 h，能够以整体粒子的形式吸收，并主要分布在肝脏。本项目发表相关学术论文10篇，其中SCI收录论文5篇，申请国际发明专利（PCT）1项，国家发明专利1项。