新型雷公藤内酯醇衍生物抑制TAK1-JNK通路调控巨噬细胞极化抗类风湿性关节炎的机制研究

类风湿性关节炎等自免病仍然是一类病因复杂、机制不明的难治性疾病，基于有效的传统中药发现新型抗自免病天然产物或衍生物将起到事半功倍的作用。传统中药雷公藤对类风关等自免病有良好的治疗效果，但同样显著的副作用限制了其更广泛的临床应用。前期研究在雷公藤内酯醇的基础上得到了一个抗炎作用明确的新衍生物ZT01，其毒性只有雷公藤内酯醇的1/160，有非常好的成药前景；初步的研究显示ZT01对TAK1磷酸化有明显的抑制作用，同时调控巨噬细胞表型极化，但其确切的靶点和作用机制还不明确。本项目拟进一步探讨ZT01作用于TAK1的确切靶点；通过考察ZT01作用于TAK1后对下游MAPK和NF-κB通路的调控机制，阐明其可能的抗炎作用机制；比较野生型和条件敲除TAK1或JNK情况下ZT01对巨噬细胞表型极化的调控机制。本研究不仅有助于深入了解自免病发病机制和发现新的靶点，也为抗自免病的新药研发提供理论基础。

Autoimmune diseases such as rheumatoid arthritis (RA) are kinds of diseases with diverse mechanisms of action, due to the complex of pathogeny, there is a clear unmet clinical need for new effective drugs. Owing to the development of novel natural products or derivatives from traditional Chinese medicine, there is good opportunity for the study of derivatives from traditional Chinese medicine on the treatment of autoimmune diseases. The Chinese herb Tripterygium wilfordii Hook F (TWHF) has been used in traditional Chinese medicine for the treatment of autoimmune diseases including RA, one major active and toxic component isolated from TWHF is triptolide (TP). In clinical settings, widespread reports shown that the application of TP is associated with diverse toxic effects. Over the past few decades, great effects have been made to explore new derivative of TWHF with higher efficacy and lower toxicity. Here, we describe a novel derivative of triptolide, ZT01, a potent and promising anti-inflammatory candidate. The toxicity of ZT01 is 1/160 of triptolide. Our previous study finds that ZT01 can selective inhibit the phosphorylation of TAK1, meanwhile, it can regulate the polarization of macrophage. However, the exact target and molecular mechanisms need further study. The aim of this project is to study the target of ZT01 on TAK1-TAB1 complex using pull-down, Co-IP, SPR, FRET and so on. Besides, the molecular mechanism of ZT01 will be further demonstrated by gene silencing and locus mutation. As reported from literatures, TAK1 is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family and one of the most important regulatory components in NF-κB signaling pathway. In the project, we will explore the anti-inflammatory mechanisms of ZT01 on the phosphorylates of IKK complex as well as p38, c-Jun N-terminal kinase (JNK), and extracellular signal regulated kinase (ERK), thus regulate the downstream signaling pathway NF-κB and MAP kinase. Through gene knockout of TAK1 in macrophage to find the mechanism of ZT01 on macrophage polarization. This finding will provide detail evidence for the mechanism of autoimmune diseases and novel drug targets, more importantly, help to the fundamental basis for new drug development on autoimmune diseases.