Clonostachys rogersoniana发酵改性香青藤的化学机制及抗肿瘤活性物质基础研究

Our previous study revealed that rhizosphere soil fungi Clonostachys rogersoniana could facilitate the biotransformation of (R)-dicentrine to (4R,6aR)-4-hydroxyldicentrine, which exhibits the same level of cytotoxicity as (R)-dicentrine, but possesses much higher water solubility than (R)-dicentrine. Fermentation of Illigera aromatic (folk medicine of the Zhuang nationality) with C. rogersoniana also led to the biotransformation of actinodaphnine to 4R-hydroxylactinodaphnine, whose water solubility was significantly increased. Interestingly, a novel diterpene named rogernine, which exhibited significant cytotoxicity, was isolated from fermented I. aromatic. Previous studies suggested that rogernine was derived from the co-effect of fungus and plant materials. In this project, fermentation of I. aromatic with C. rogersoniana will be studied carefully. It is hopeful to find more 4R-aporphine alkaloids and rogernine analogues and develop (high water-soluble) antitumor lead compounds. According to systematic fermentation research, the chemical mechanism of the production of rogernine will be investigated. This study will facilitate the discovery and mass production of 4R-hydroxylaporphine alkaloids and rogernine, clarify the relationship between I. aromatic and C. rogersoniana during fermentation process. This project will offer a new thought for the study of fermented traditional Chinese medicine.

申请人前期研究发现根际土壤真菌Clonostachys rogersoniana能定量的将荷包牡丹碱转化成抗肿瘤活性相当而水溶性更高的4R-羟基荷包牡丹碱。采用该菌发酵壮族民间药香青藤时，发现该菌同样能定量的将香青藤中的放线瑞香宁转化成高水溶性的4R-羟基放线瑞香宁。最有趣的是在发酵香青藤中还发现了一个具有较强抗肿瘤细胞活性的新颖二萜rogernine，初步研究表明该化合物由微生物和植物共同产生。本项目拟深入研究C. rogersoniana对香青藤的发酵改性，分离更多的4R-羟基阿朴菲生物碱及rogernine类似物，发现（高水溶性）抗肿瘤活性化合物，并通过系统发酵研究探讨rogernine产生的化学机制。该研究有望实现抗肿瘤活性先导化合物4R-羟基阿朴菲生物碱和rogernine的发现和量产，阐明发酵过程中C. rogersoniana与香青藤的关系，为发酵药材研究提供新思路。

青藤属植物研究较少，本项目开始之际，国内青藤属植物只有吕宋青藤、黑吹风和香青藤有少量报道。青藤属植物中主要的化学成分是阿朴菲类生物碱，而青藤属药用植物的药理作用多与其所含阿朴菲类生物碱有关。本项目从四种青藤属植物：香青藤、蒙自青藤、大花青藤、心叶青藤，两株宽叶青藤植物内生真菌：Gliocladiopsis SP. A8、Pseudopestalotiopsis SP. K7，罗杰斯无性穗霉（Clonostachys rogersoniana）中总共分离了104个化合物，其中25个为新化合物，包括二聚䓝烷类化合物、阿朴菲类生物碱和萜类化合物，部分化合物具有良好的抗肿瘤细胞活性、乙酰胆碱酯酶抑制活性和抗菌活性。探索了罗杰斯无性穗霉产生抗肿瘤活性化合物TMC154的化学机制。研究了罗杰斯无性穗霉对香青藤和蒙自青藤的微生物发酵改性，发现了3个新的微生物转化反应。发现产生4R-羟基阿朴菲生物碱的生物转化反应，阐明了二聚䓝烷类化合物产生的化学机制，其来源实际为酯类化合物水解。本项目为青藤属药用植物的开发提供了重要的科学依据。