前体物调控激素自养型蛇足石杉叶状体转化Hup-A的方法及分子基础研究

Huperzine A (Hup-A) preparation is the most effective drug to treat Alzheimer's disease (AD) at home and abroad currently, especially Hup-A derived from plant Huperzia serrata could effectively fight against AD. However, the biggest bottleneck to develop new drugs is the lack of H.serrata resources. Applicant established a hormone autotrophy Huperzia serrata clonal thallus in vitro platform to produce Hup-A and found that there is conversion phenomena between the reduction of secondary metabolic fingerprint T intermediate and the increase of Hup-A in the thallus added with a precursor compound. Depending on the principle of the plant, secondary metabolites can be adjusted by multiple ways. With hormone autotrophy Huperzia serrata tallus in vitro as object, this project aims: (1) To study the law of different methods, time and concentration of added precursor compounds and their biological response and regulation of generating Hup-A from the fingerprint T intermediate product; and to isolate and identify the fingerprint T intermediate monomer; (2) To research the law of the vitro tallus proliferation and the endogenous hormones level changing and the relationship between Hup-A produced by secondary metabolism and Related to specific protein; (3) To compare transcriptome of several typical leafy phenotype/strains, and to reveal the differences between transcriptional gene cluster, related functional genes or related biochemical system of generating Huperzine by transciptome RNA-seq high-throughput sequencing and bioinformatics analysis. The ultimate goal of this project is to construct artificial metabolic engineering to efficiently regulate the production of Hup-A by additive precursor and molecular mechanism of Hup-A produced by Huperzia serrata. There are important scientific significance and appliance value to protect unique Chinese plant Huperzia serrata and to maintain renewable sources of Hup-A drugs.

石杉碱甲（Huperzine-A即Hup-A）制剂是目前国内外最为成功治疗老年性痴呆症的药物，尤其是来源于植物源蛇足石杉的Hup-A具有活性高的药效，但此药源匮乏已成为新药研发的最大瓶颈。申请者通过前期工作，建立了Hup-A的激素自养型蛇足石杉离体叶状体培养的繁殖系平台，并发现添加一种前体化合物培养叶状体，次生代谢的指纹T中间产物消减的同时Hup-A增多的转化相关现象。依据植物次生代谢多途径可调控等原理，本项目拟基于激素自养型蛇足石杉离体叶状体为对象，研究前体化合物添加的方式、时间、浓度的生物技术调控响应及其指纹T中间产物转化生成Hup-A的规律，分离鉴定指纹T中间产物等单体，探索离体叶状体次生代谢的相关内在联系。揭示前体物调控蛇足石杉叶状体转化Hup-A的分子基础和提出转化提高Hup-A含量的关键技术，它将在工厂化持续再生植物源Hup-A药物资源应用上具有重要的价值和科学意义。

石杉碱甲（Huperzine-A即Hup-A）制剂是目前国内外最为成功治疗老年性痴呆症的药物，尤其是来源于植物源蛇足石杉的Hup-A具有活性高的药效，但此药源匮乏已成为新药研发的最大瓶颈。本项目基于激素自养型蛇足石杉离体叶状体离体培养为基础，研究了前体化合物添加的方式、时间、浓度的生物技术调控响应及其叶状体代谢相关的生化特征；鉴定了伴随HupA累积的相关三种次生代谢物质及其物质间累积的可能关系。揭示了前体物调控蛇足石杉叶状体转化Hup-A的分子基础和提高培养物Hup-A含量的关键技术，它将在工厂化持续再生植物源Hup-A药物资源应用上具有重要的价值和科学意义。