基于新型无标记细胞筛选技术的辽宁道地药材药效物质基础研究

Liaoning Province can field a score of genuine medicinal material, because of the influence from Changbai Mountains. How to carry out deep study about these herbs in order to contribute to the economic development and TCM companies’ transformation of Liaoning? In this project, based on the high efficient separation technology and new label-free cellular integration pharmacology which were established by ourselves, the applicants will give a systematic research about 20 kinds of Liaoning Province genuine medicinal material in order to find the active substance with clear targets . In an extensive screening, more than 3000 fine fractions from 20 herbs will be screened in 20 kinds of GPCR targets which are orphan GPCR or related to cardiovascular, cancer, allergies diseases. With the development of efficient separation and purification methods of the active fractions, the active compounds with clear targets will be found and become leading compounds for the relevant disease. Since 9 herbs have been reported to be analgesic, they were focused on the analgesic mechanism research. The selective pretreatment methods were established to obtain fraction rich with alkaloids, saponins, lactones or terpene compounds, meanwhile a 15 kinds of GPCR targets modal associated with pain will be established including different subtypes receptors of dopamine, opioids, 5-hydroxy tryptamine. Fractions rich with same kind of compounds will be screened and active compounds will be found. Both active fractions an compounds lay a foundation for the development of analgesic clinical TCM .

辽宁省因属长白山的余脉，是二十余种道地药材的产地。如何有针对性地对这些道地药材开展深入研究，从而更好地为辽宁的新兴经济建设和中药企业转型做出贡献？本项目针对上述问题，利用申请者在高效纯化分离技术和新型无标记细胞整合药理学方面的基础，从广度和深度两方面，对20种辽宁道地药材开展活性物质和作用靶点的系统研究。在广度上，完成3000个精细组分，在心血管、癌症、过敏、孤儿GPCR等20种靶点模型上的活性评价，并发展高效的活性化合物追踪技术，发现具有明确作用靶点的活性物质，为相关疾病提供先导化合物。在深度上，探究具有镇痛、镇静作用的9种道地药材的药效物质基础，建立选择性预处理方法，获得药材中生物碱、皂苷、内酯、萜类等类化合物组分，建立镇痛相关的不同亚型多巴胺、阿片、5-羟基色胺等15种GPCR筛选模型，发现具有活性的类化合物组分或化合物，结合大鼠药效学实验，为临床开发镇痛中药奠定基础。

辽宁省因属长白山的余脉，是二十余种道地药材的产地。如何有针对性地对这些道地药材开展深入研究，从而更好地为辽宁的新兴经济建设和中药企业转型做出贡献？本项目针对上述问题，利用申请者在高效纯化分离技术和新型无标记细胞整合药理学方面的基础，对辽宁道地药材开展活性物质和作用靶点的系统研究。基于新型无标记细胞整合药理学技术，构建了疼痛、神经退行性疾病、心血管疾病、代谢性疾病、慢阻肺、肿瘤等疾病相关的42种GPCR靶点模型的构建，包括阿片类、毒蕈碱类、多巴胺类、肾上腺素类及五羟色胺类等受体，打破了中药机理研究模型单一的局限性；完成了22种道地药材的400个精细组分在疼痛、心血管、癌症重点相关的32种靶点模型上的活性评价，发现了50个靶点活性明确的精细组分；并发展了高效的活性化合物追踪技术与方法，发现了19个结构明确的靶向活性化合物，其中发现2个化合物具有阿片受体的激动活性，为镇痛药物研发提供先导化合物；基于发现的具有GPR35靶向活性的双香豆素，开展了68个化合物的设计与合成，发现了纳摩尔水平的高活性的激动剂，为新药开发提供了候选分子。鉴于辽宁中药产业的升级，项目增加了辽宁道地药材的质量标准化研究与绿色制造工艺研究，形成了辽宁中药的整体质量控制，开发了系列高品质的精制配方颗粒。本项目系统的研究了辽宁道地药材的药效物质基础、质量标准与绿色制造工艺，为中药新药研究奠定了科学基础，为促进辽宁中药产业发展提供了有效技术。