喜树定向培育及喜树碱衍生物水溶性制备工艺创新

本项目针对喜树碱和羟基喜树碱两种抗癌生物碱在我国特有植物喜树中含量低，传统提取工艺仅以喜树种子为原料，而丰量的含有喜树碱衍生物的叶片未得以充分利用以及生产出的喜树碱衍生物水溶性差、生物利用度低、毒副作用大，达不到国际高端原料的质量标准的产业现状，以提高我国喜树资源的定向培育、提取加工及终端原料药质量标准的产业化生产工艺为研究目的，在喜树定向培育方面，通过组培选育出枝叶采收利用型喜树无性系，并以生活史型理论为指导，采用人工复合群落定向培育技术诱导喜树中喜树碱的生物合成 在喜树碱衍生物的提取分离方面，以喜树叶片替代传统的喜树种子为原料，采用匀浆萃取、连续中压柱层析等技术提高喜树碱的纯度和得率，达到国际原料药的质量标准 在水溶性喜树碱衍生物制备方法，基于胶体化学理论和纳米技术，研发出水溶性喜树碱衍生物粉体制备技术与装备，使喜树碱衍生物达到国际高端原料药的质量标准。通过运用上述创新工艺和技术，本项目通过组培选育出喜树碱高产无性系，其喜树碱含量比普通喜树高出15%-30% 采用营造喜树、南方红豆杉、迷迭香人工复合群落，并通过控制人工复合群落内环境因子定向诱导喜树中喜树碱的生物合成，使喜树碱的含量较目前野生型喜树提高80.3% 并通过陈化处理使叶片中喜树碱和羟基喜树碱的含量分别由0.8‰、0.1‰增量为0.85‰和0.76‰，解决了喜树碱衍生物生产中的新原料问题。以喜树叶片替代传统的喜树种子进行喜树碱衍生物分离，在分离过程中采用匀浆萃取、连续中压柱层析等技术，使喜树碱纯度由92%提高至98%，羟基喜树碱纯度由90%提高至98%，均达到国际原料药的质量标准 并基于胶体化学理论和纳米技术，研发出水溶性喜树碱衍生物粉体制备技术与装备，水溶性喜树碱和羟基喜树碱的平均粒径均不超过200nm，溶解度较原粉分别提高23.9 和11.2 倍，体内抑瘤效果显著优于原药，毒性作用明显降低，为我国喜树碱类新制剂的开发提供了新型原料药品种。

This project addresses the current industrial status quo where two anticancer alkaloids, camptothecin (CPT) and hydroxycamptothecin (HCPT), exist in low contents in Camptotheca acuminata, a plant endemic to China; traditional extraction processes only use camptotheca seeds as raw materials, leaving abundant leaves containing camptothecin derivatives underutilized; additionally, the produced camptothecin derivatives suffer from poor water solubility, low bioavailability, and severe adverse reactions, failing to meet the quality standards of international high-end raw materials. The research objective of this project is to advance the industrial production processes for targeted cultivation of China’s camptotheca resources, extraction and processing, and quality standardization of terminal bulk drug substances. For targeted camptotheca cultivation, we screened and bred foliage-harvesting and utilizable camptotheca clonal lines via tissue culture. Guided by the life history theory, we adopted artificial mixed community targeted cultivation technology to induce camptothecin biosynthesis in camptotheca. Through these efforts, we obtained a high-yield camptothecin clonal line with a camptothecin content 15%-30% higher than that of ordinary camptotheca. Furthermore, by constructing artificial mixed communities of camptotheca, Taxus chinensis var. mairei, and Rosmarinus officinalis, and controlling environmental factors within the communities to directionally induce camptothecin biosynthesis, the camptothecin content was increased by 80.3% compared to current wild-type camptotheca. Additionally, aging treatment increased the contents of camptothecin and hydroxycamptothecin in leaves from 0.8‰ and 0.1‰ to 0.85‰ and 0.76‰ respectively, resolving the raw material shortage issue in camptothecin derivative production. For extraction and separation of camptothecin derivatives, we replaced traditional camptotheca seeds with camptotheca leaves as raw materials, and adopted technologies including homogenization extraction and continuous medium-pressure column chromatography to improve the purity and yield of camptothecin, enabling the products to meet international bulk drug substance quality standards. Specifically, using camptotheca leaves instead of seeds for separation, the purity of camptothecin was increased from 92% to 98%, and the purity of hydroxycamptothecin was increased from 90% to 98%, both meeting the quality standards of international bulk drug substances. For the preparation of water-soluble camptothecin derivatives, based on colloid chemistry theory and nanotechnology, we developed a preparation technology and equipment for water-soluble camptothecin derivative powders. The average particle sizes of water-soluble camptothecin and hydroxycamptothecin were both no more than 200 nm, their solubility was increased by 23.9 and 11.2 times respectively compared to the original powder, the in vivo tumor-suppression effect was significantly better than that of the original drug, and toxic side effects were markedly reduced. This work provides a novel bulk drug substance variety for the development of new camptothecin-based preparations in China.

搜集汇总

背景与挑战

背景概述

该数据集详细介绍了喜树定向培育技术及喜树碱衍生物水溶性制备工艺的创新方法，包括通过组培选育高产无性系、人工复合群落定向培育技术、以及基于胶体化学和纳米技术的水溶性制备工艺。这些创新显著提高了喜树碱的产量、纯度和水溶性，为抗癌药物的研发提供了高质量的原料药。