基于NO抑制活性的四种云南产姜花属植物化学成分及其细胞毒活性研究

A large number of species from the genus of Hedychium(Zingiberaceae)are distributed in China and about 85% of them were found in Yunnan province, including 14 species endemic to Yunnan, which stands out prominently in natural resources compared with other provinces of China. However, not enough research papers deal with chemical constituents or active compounds from species of the genus of Hedychium. In recent years, we investigated the chemical constituents from the species of Zingiberaceae in order to find active constiuents with inhibition on production of nitric oxide（NO）and 90 compounds were isolated, including 17 novel sesquiterpenes, 8 new sesquilignans and 5 new diterpenes. Inhibitory activities of 45 compounds were tested against NO prodction in interferon-γ(IFN-γ)- and lipopolysaccharide(LPS)- induced RAW264.7 macrophage cells. Among them, 31 compounds showed different activity and IC50 values of them ranged from 4.93 to 13.95 μg/ml. As a part of our ongoing project towords the discovery of novel biologically active metablites from the plants of Zingiberaceae, 8 species from the genus of Hedychium were evaluated for their NO production inhibitory activity and the methanol extracts of H. sinoaureum, H. coronarium, H. longpetalum and H. simoense exhibited inhibitory effects with IC50 values from 8.66 to 13.78 μg/ml. Further, these plants showed singnificant cytotoxicities against three cancer cell lines with IC50 values from 8.09 to 17.28 μg/ml. Next, the secondary metabolites from the above 4 species and their NO production inhibitory activities and cytotoxicities will be revealed in this project. Finally, the structure-activity relationships of their activities will be discussed. Results of this project will be help to release the mechanism of inhibition of NO production and cytotoxicities, and to find new pharmacological activities for the species due to the relationships of NF-κB and NO production with some diseases including cancer, and to exploit medicinal plants of the genus of Hedychium in Yunnan province.

滇产姜科姜花属植物资源丰富，约占全国的85％，且多种植物为云南特有种，但其化学活性成分研究不多。为了深入研究姜花属植物活性成分，申请人已从多种姜科植物分离到化学成分90余个，包括17个新颖倍半萜、8个新倍半木脂素、5个新二萜，其中31个成分显示了较强的NO抑制活性，其IC50值范围为4.93-13.95 μg/ml。并且申请人对姜花属8种植物进行了NO抑制活性测试，结果显示4种植物的活性较强，其IC50值范围是8.66-13.78μg/ml；且它们对三种肿瘤细胞株有较强的细胞毒活性，其IC50值范围在8.09-17.28μg/ml。由于NF-κB通路与NO的合成以及肿瘤的发生有密切关系，本项目拟从这4种植物中筛选出重要的NO抑制剂，同时以该结果导向性筛选细胞毒活性分子，并阐明其构效关系。本项目的实施为姜花属植物药理作用机制和新的药理活性研究奠定物质基础。

一氧化氮（NO）是近年来发现的一个具有重要生物学意义的生物调质，存在于多种细胞中。机体内NO的合成及其生理浓度与多种疾病的发生有密切的关系，尤其是与肿瘤细胞毒和炎症的发生机制相关。项目以NO抑制活性和细胞毒活性较好的6种滇产姜花属植物（小花姜花、姜花、长瓣裂姜花、思茅姜花、滇姜花、草果药）为对象，研究了姜花属植物的次生代谢产物，并进行了抑制巨噬细胞NO产生的活性和细胞毒活性筛选。其研究结果如下：.（1）项目以NO抑制活性和细胞毒活性较好的6种滇产姜花属植物（小花姜花、姜花、长瓣裂姜花、思茅姜花、滇姜花、草果药）为研究对象，分离化学成分122个，鉴定了其中的89个，发现新化合物10个。.（2）对分离的化合物进行了NO抑制活性或细胞毒活性筛选工作，其中12个化合物巨噬细胞中NO的产生又较好的抑制作用，其IC50的范围为：0.56-7.50μg/ml；10个化合物对人宫颈癌细胞（Hela）产生又较好的抑制作用，其IC50的范围为：3.75-14.83μg/ml；10个化合物对人胃癌细胞（SGC-7901）产生又较好的抑制作用，其IC50的范围为：2.86-14.88μg/ml；通过构效关系研究发现半日花烷型二萜C-12处的内酯环消失，活性也会大幅减弱或消失。.（3）项目发现7个化合物既有NO抑制活性，也有肿瘤细胞毒活性，其药理活性作用机制可能与NF-κB信号通路和一氧化氮合酶相关。.（4）长瓣裂姜花、思茅姜花是上世纪九十年代以后发现的姜花属新种，现代的植物化学研究方面是空白，本研究首次揭示其内在药理活性化学成分，其研究结果为其药理作用机制和今后的开发打下物质基础。.（5）发表学术论文11篇，其中4篇为SCI收录论文。.本项目揭示了6种滇产姜花属植物的次生代谢活性成分，发现了多个药理活性成分，其中长瓣裂姜花、思茅姜花是上世纪九十年代以后发现的姜花属新种，项目首次揭示了其药理活性成分。另外，NO与多种临床疾病尤其是与肿瘤发生的密切相关性，使本研究为该属植物NO抑制活性及肿瘤细胞毒药理作用机制奠定物质基础，对其它新的药理活性研究具有重要的启示作用，也可为合理、可持续开发利用丰富的滇产姜花属植物资源和云南特色创新药物研制提供科学依据。