新型活性单体芍药苷-6-氧-苯磺酸酯在类风湿关节炎免疫应答与炎症中的调控作用

Total glucosides of paeony (TGP) is the first anti-inflammatory immune regulatory drug in treatment for rheumatoid arthritis (RA) approved in China. New active monomers paeoniflorin - 6 - oxygen - benzene sulfonic acid ester (code is CP - 25) come from the structure modification of paeoniflorin (Pae), which was the effective active ingredients of TGP, and the patents of CP – 25 has been applied. Compared with Pae, CP – 25 has good anti-inflammatory and immune regulation activity, and the absolute bioavailability of CP – 25 is high. In RA immune response and synovial inflammation, it was found that dendritic cells (DCs) activated T cells by presenting the antigen to T cell, the interaction between T cell and B cell or FLS, and B cells were activated, inflammatory change of FLS was induced. how does CP - 25 regulate immune response and synovial inflammation in RA? Whether does CP – 25 play its regulation function through the adjustment of DCs, T and B cells and FLS function and cell interaction? No reports in the literature have seen at home and abroad. In this project, RA patients and animal model intend to be as research object, agonist, antagonist and siRNA will be used as intervention, and different molecular biology technology will be utilized. The regulation of CP - 25 on the functions and cell interaction of DCs, T and B cells and FLS will be investigated, and the regulation mechanism of CP - 25 will be revealed. This project will offer a solid foundation for developing CP - 25 as the independent intellectual property rights of new original drugs.

白芍总苷（TGP）是我国第一个批准上市治疗类风湿关节炎（RA）抗炎免疫调节药。新型活性单体芍药苷-6-氧-苯磺酸酯（代号CP-25）来自于TGP有效活性成份芍药苷（Pae）结构修饰，已申请专利。与Pae相比，CP-25绝对生物利用度高，具有良好抗炎和免疫调节活性。树突细胞（DCs）递呈抗原活化T细胞，T细胞与B细胞和FLS相互作用，活化B细胞，诱导FLS炎性转变，参与RA免疫应答和滑膜炎症。CP-25对RA免疫应答和滑膜炎症具有怎样调控作用？是否通过调节DCs、T、B细胞和FLS功能及细胞间相互作用发挥其调控作用？国内外未见文献报道。本项目拟以RA患者和动物模型为研究对象，通过激动剂、拮抗剂和siRNA等干预，采用不同分子生物学技术，探讨CP-25对DCs、T、B细胞和FLS功能及细胞间相互作用的调节，并揭示CP-25调控机制。为把CP-25开发为独立自主知识产权新的原创药物打下坚实基础。

新型活性单体芍药苷-6-氧-苯磺酸酯（代号CP-25）来自于白芍有效活性成份芍药苷结构修饰，具有良好抗炎和免疫调节活性。树突细胞（DCs）递呈抗原活化T细胞，T细胞与B细胞和成纤维滑膜细胞（FLS）相互作用，参与RA免疫应答和滑膜炎症机制尚不清楚。本项目以RA患者组织样本和RA 动物模型为研究对象，考察CP-25 对免疫细胞影响，考察CP-25对免疫细胞相互作用、免疫细胞和炎性FLS相互作用的影响及分子机制，研究CP-25对免疫细胞G蛋白偶联受体（GPCRs）介导的信号通路在上述病理环节中的变化以及活性调控靶点。研究发现CP-25 对佐剂性关节炎大鼠、胶原性关节炎(CIA）大鼠和 CIA 小鼠等模型具有明显的治疗作用，可以降低动物模型外周血炎性细胞因子水平和自身抗体水平，调节外周血总 B 细胞、TACI+B 细胞、活化 B 细胞、BAFFR+B 细胞和浆细胞比例恢复至接近正常水平，调节外周血调节性 T 细胞和效应性 T 细胞及其亚群恢复至生理水平。CP-25对RA 患者外周血淋巴细胞和RA患者成纤维滑膜细胞株异常活化具有显著的调控作用，对关节炎动物模型异常活化的T细胞、B细胞、DC、FLS细胞及其相互作用具有显著的调控作用。CP-25可以调控上述免疫细胞和滑膜细胞上前列腺素受体、趋化因子受体、β-肾上腺素受体等介导的信号转导通路恢复至正常水平，可以抑制多个GPCRs介导信号转导通路与BAFF 及其受体介导的信号转导通路的交叉活化。进一步研究发现CP-25具有炎症免疫反应软调节作用，这与其作用G蛋白偶联受体激酶-2（ GRK2 ）激酶活性区的氨基酸残基、调节GRK2活性，调控GPCRs 异常脱敏及其信号通路异常有关。本研究明确了GPCRs介导的信号通路异常是类风湿关节炎发病新的病理机制，发现GRK2 是类风湿关节炎治疗药物的新靶点，这为临床治疗类风湿关节炎及其研发创新治疗药物提出了新思路。