基于中药配伍理论的南天竹组分配伍对抗肿瘤药三氧化二砷减毒机制研究

Results of the project (No. 81102788) supported by the National Science Foundation showed: 1) the extract of Nandina domestica had significant thetoxicity-reducing and activity-maintaining effects on arsenic triode (See our patent:ZL201110089819.6); 2) the alkaloids and phenolics were the active constituents of protection of Nandinadomestica against cardiotoxicity and nephrotoxicity induced by arsenic triode, respectively; 3) the attenuated toxicity effect of single compounds and the chemical fractions were worse than that of the extraction, and the effect of the composion of fourteen main alkanoids (about 85% total alkanoids of Nandina domestica) and ten phenolic acids (about 80% total phenolic acids) was equal to that of the exaction.It suggested that two ingredients of the alkaloid and phenolic acid from Nandina domestica had present the compatibility effects on detoxification of arsenic triode, and possessed TCM characteristics, such as multi ingredients, targets and ways. However, the definite active components are not known. The proportion of every compound in the active components is unkown. The mechanism of toxicity-reducing activity of the active components is unclear. So, in this study, we will indentify the active components group by using methods of the stepwise multiple regression analysis and the baseline geometric proportion increasing and decreasing design, combinated the results of activities. We will elucidate the components of the toxicity-reducing activity, clarify the relation of quantity and effectiveness among components. Using methods of the metabonomics, we will investigate the characters of “Jun”, “Chen”, “Zuo” and “Shi” of active components, and attempt to elucidate the mechanism of the toxicity-reducing activity. So this study can improve the safety of arsenic triode of clinical apply, and can provide the research base of the new medicine of anti-tumor.

上个基金（81102788）研究显示南天竹对As2O3抗肿瘤减毒持效作用具多成分、多靶点、多途径中医药特点：生物碱和酚酸分别是拮抗心和肝肾毒主要活性成分；单体及化学部位效果均不及总提物；由占总生物碱85%的14个生物碱和总酚酸80%的10个酚酸组成的组分与总提物减毒持效作用相当。然而，上述组分中确切的有效组分尚不清楚，有效组分的最优配比仍然未知，有效组分间各成分协同作用机制仍不清楚。据此，本课题依据中药配伍理论，采用逐步多元回归、均匀设计和网格法，结合减毒持效药效评价，剔除“相畏和相恶”成分，优化“相须和相使”成分配比，进而阐明拮抗As2O3毒性有效组分，明确有效组分间量效关系。应用代谢组学探讨组分间 “君臣”与“佐使”，揭示协同减毒机制。项目研究结果为提高As2O3和含As2O3药物安全性提供新方法，为研发毒副作用小、安全性能高、成分机理明确的As2O3抗肿瘤复方新药奠定基础。

南天竹对As2O3抗肿瘤具有减毒持效作用肯定，具多成分、多靶点和多途径中医药特点。由生物碱和酚酸主成分的组分与总提物对As2O3减毒作用相当，而上述组分中确切组成不清，最优配比未知，组间各成分协同作用机制不明。本项目旨在解决上述问题，开展了以下研究：有效组分的辨识；有效组分配伍及配比优化探讨；有效组分协同减毒机制研究。得到以下结果：①“相使或相须”成分：南天竹碱、南天竹宁碱、N-甲基南天宁碱、小檗碱、没食子酸、1-羟基松脂素、杜仲树脂酚、berchemol-4'-O -葡萄糖苷和丁香脂素；“相畏或相恶”成分：去氢南天宁碱、南天表碱、药根碱、4,5-二氧代去氢南天宁碱、海罂粟碱、去氢紫堇碱、邻苯二甲酸二丁酯和咖啡酸；“杂质”成分：木兰碱、O-甲基球紫堇碱、nandsterine、巴马亭、原儿茶酸、丁香酸和对羟基桂皮酸；②“相使或相须”成分配比为生物碱部分:酚酸部分3:1，其中生物碱中南天竹碱、南天竹宁碱、N-甲基南天宁碱与小檗碱比为2:1:3:3，酚酸中没食子酸、1-羟基松脂素、杜仲树脂酚、berchemol-4'-O-葡萄糖苷和丁香脂素2:3:3:1:2；③有效组分中“君-臣”成分：南天竹碱、南天竹宁碱、N-甲基南天宁碱、小檗碱、1-羟基松脂素、杜仲树脂酚和丁香脂素；“佐-使”成分：没食子酸和berchemol -4'-O-葡萄糖苷；④“佐-使”成分显著地影响“君-臣”成分的吸收和代谢；有效组分对As2O3诱导脏器损伤具有保护作用，对CYP450酶活力产生抑制。通过本项目的实施，完成了项目研究内容，实现了预定目标，培养了5名硕士生，3名本科生；发表论文6篇，申请2项专利，3篇正整理数据和撰写。另外，受本项目资助开展的研究内容已发表6篇论文（已标注）。研究结果为临床应用提高As2O3的安全性提供新方法，进而为研发毒副作用小、成分与机理明确的As2O3抗肿瘤复方新药提供理论基础。