花椒属植物中PD-1/PD-L1信号通路小分子抑制剂的挖掘及功能评价

Checkpoint blockers or drugs targeting PD-1 and its ligand interactions are the focus of current research on tumor immunotherapy. At present, there is no clinical application of small molecule inhibitors for this pathway except monoclonal antibodies. The existence of amide functional groups in the artificially synthesized small-molecule inhibitors are the basis for its binding to specific amino acid sites of PD-1 or PD-L1. As edible Chinese herbals, the genus Zanthoxylum are rich in amide compounds and can be a natural source of small-molecule inhibitors of PD-1/ PD-L1 pathway with low toxicity. The extract of Sichuan pepper and its EtOAc fractions showed a strong inhibitory effect of PD-1/ PD-L1 pathway in the ELISA test. In this study, extracts of the genus Zanthoxylum plants will be screened, and effective parts will be subjected to phytochemical investigation. The inhibitory effect of the amides in PD-1/ PD-L1 pathway and their structure-activity relationships were evaluated on the basis of SYBYL molecule predicts and ELISA assay. Furthermore, melanoma animal model was used to evaluate the antitumor effect of these compounds in vivo. This study looks forward to obtain efficient small-molecule inhibitors of PD-1/ PD-L1 pathway and provide new candidate drugs for the immuno-blocking therapy in tumor immunotherapy.

针对PD-1/PD-L1间相互作用的免疫检查点阻断技术是当前肿瘤免疫疗法研究的热点。目前除单克隆抗体外，尚无此通路有效小分子抑制剂的临床应用。人工合成的PD-1/PD-L1小分子抑制剂中酰胺官能团的存在是其与PD-1或PD-L1特定氨基酸位点相结合的基础。药食两用的花椒属植物因富含酰胺类化合物可作为高效低毒的PD-1/PD-L1通路小分子抑制剂的天然来源。申请人通过ELISA实验评价模型确定了花椒提取物及其乙酸乙酯部位具有很强的PD-1/PD-L1通路抑制作用。本项目拟在此基础上对花椒属植物酰胺类化合物进行广泛筛选和分离，基于SYBYL分子预测和ELISA实验评价酰胺类成分的PD-1/PD-L1通路抑制作用及其构效关系，并进一步使用小鼠黑色素瘤模型评价这类化合物的体内抑瘤作用。以期获得针对PD-1/PDL1通路的小分子抑制剂，为肿瘤免疫疗法中的免疫阻断治疗技术提供新候选药物。

针对PD-1及其配体间相互作用的免疫检查点阻断方法或药物是当前肿瘤免疫疗法研究的热点。目前除单克隆抗体外，尚无此通路有效小分子抑制剂的临床应用。人工合成的PD-1/PD-L1小分子抑制剂中酰胺官能团的存在是其与PD-1或PD-L1特定氨基酸位点相结合的基础。药食两用的花椒属植物因富含酰胺类化合物可作为高效低毒的PD-1/PD-L1通路小分子抑制剂的天然来源。.本研究采集了花椒属其他种药材的不同药用部位近100种，对各药用部位进行95%乙醇冷浸提取。并利用已建立的ELISA评价体系，对不同药用部位95%乙醇提取物进行PD-1/PD-L1相互作用抑制活性评价。从不同药用部位95%乙醇提取物进行PD-1/PD-L1信号通路抑制活性来看，花椒属植物部分种的药用部位存在一定的抑制作用，从药用部位来看，根部及果皮的部位显示出相对较好的活性。基于前期实验结果，本项目对花椒属植物簕欓花椒根进行化学成分研究，并通过多种色谱分离方法从簕欓花椒95%乙醇冷浸提取物中分离得到单体化合物，并对目前花椒属植物来源的单体化合物进行了多种生物活性定。