基于Hsp90的乌头二萜生物碱构效关系与靶向抗肿瘤毒效机制研究

Diterpenoid alkaloids, the characteristic components of genus Aconitum, have strong toxicity and various biological activities such as analgesia and anti-tumor. Our previous studies found their toxicities are closely related to the molecular approximate surface area and dipole and other structural parameters. Moreover, molecular docking showed that heat shock protein 90 (Hsp90) may be the potential anti-tumor target. The project is proposed on the basis of previous research work. With diterpenoid alkaloids as lead compounds, dominant pharmacophores are extracted by using protein-ligand interaction fingerprint (PLIF) analysis method based on the docking conformations of diterpenoid alkaloids and Hsp90 in order to increase anti-tumor activity. Furthermore, the strategies such as destroying rigid structure, introducing water-soluble groups, improving the molecular flexibility and polarity will be adopted in order to achieve the purpose of reducing toxicity. The structure of diterpenoid alkaloids will be transformed and optimized by using a computer aided drug design method. Novel diterpenoid alkaloid derivatives will be synthesized. Their toxicities and anti-tumor activities will be evaluated in vitro and in vivo, and the mechanisms of targeting Hsp90 will be studied. Quantitative structure-activity/toxicity relationship (QSAR) will be analysed and 4D-QSAR model will be built. Ways and means of attenuated efficiency for diterpenoid alkaloids will be researched and explored. The research will provide theoretical and experimental evidence for the application of toxic natural products and the discovery of new diterpenoid alkaloids anti-tumor drugs.

乌头属植物的特征性成份二萜生物碱具有很强的毒性，同时具有镇痛、抗肿瘤等多种生物活性。申请人研究发现该类生物碱的毒性与其分子的近似表面积、偶极距等结构参数密切相关，并且分子对接研究结果表明热休克蛋白90（Hsp90）可能是其潜在的抗肿瘤靶点。本项目拟在此基础上，以乌头碱等二萜生物碱为先导化合物，基于其与Hsp90的对接构象，利用蛋白-配体指纹图谱分析的方法，提取优势药效团，以提高其靶向抗肿瘤活性；并采取破坏刚性结构、引入水溶性基团、增加分子的柔性和极性等策略，以降低其毒性。利用计算机辅助药物设计进行结构改造和优化，制备新型二萜生物碱衍生物，进行靶向抗肿瘤毒效评价和Hsp90介导的抗肿瘤作用机制的研究。进行定量构效关系（QSAR）分析，构建4D-QSAR模型。研究并探讨二萜生物碱类化合物减毒增效的途径和方法，为毒性天然产物的应用以及新型二萜生物碱类靶向抗肿瘤药物的发现提供理论和试验依据。

本项目在前期研究工作基础上，以二萜生物碱为先导化合物，基于其与潜在的抗肿瘤作用靶点的对接构象，采用蛋白-配体指纹图谱分析的方法，提取优势药效团，并将之与毒性定量构效关系研究结果相结合，采用破坏刚性结构，引入水溶性基团，提高分子的柔性和极性，减少分子的近似表面积和疏水系数等策略，通过计算机辅助药物设计，进行结构改造和优化，降低毒性，同时增加抗肿瘤活性；设计并合成了多个系列新型结构衍生物，测定了目标化合物的体内外抗肿瘤活性和毒性，深入研究了调控毒效作用的机制；通过定量构效关系（QSAR）分析，构建4D-QSAR模型，探讨了该类化合物的减毒、增效的途径和方法，为进一步设计和筛选潜在的高活性、低毒性化合物，创制具有我国自主知识产权的靶向抗肿瘤候选药物奠定基础。在本项目研究期间，部分研究结果已发表学术论文40余篇（其中SCI论文30多篇），会议论文8篇，著作8部；申请中国专利14份，获得专利授权6份，获得国家版权局计算机软件著作权1份；获得辽宁省科技进步奖一等奖、广西药学会中恒科技进步奖一等奖、辽宁省科技进步奖二等奖和沈阳市科技进步奖二等奖各1项；培养毕业博士/硕士研究生10余名，在读博士/硕士研究生12名。