Design, Sythesis And Screening Of TNF-α Inhibitor

Human immune and certain non-immune cells can synthesize and secrete some small molecular cytokines, which modulate the biological functions of many kinds of cells. Tumor necrosis factor-a (TNF-(?)) is one of them that produced by macrophage. TNF-(?) is a multifunctional cytokine that mediates key roles in acute and chronic inflammation, anti-tumor responses and infection.The mitogen-activated protein kinases (MAPK) are important components in signal-transduction cascades that regulate diverse cellular events such as cell transformation, proliferation, differentiation, and apoptosis. MAPK family comprises four major families: the extracellular-signal regulated kinases (ERKs), Jun N-terminal kinases (JNKs), p38 MAPK and ERK5．The observation that the small molecule inhibitor of p38 MAP kinase such as SK&F86002 and SB 203580 can reduce LPS-induced TNF-(?) level in vitro and in vivo has led to numerous efforts to identify p38 inhibitors with potential for use as inflammatory diseases.Based on the structure of p38 MAPK and the interactive characteristics of ligands with p38 MAPK, a novel virtual library of TNF-(?) antagonists was constructed in a combinatorial strategy and virtual screened by DOCK 4．0 against crystal structure of p38 MAPK and BIRB-796．Two series of compounds were chosen to synthesize on the basis of the score of virtual screening.62 novel compounds have been synthesized whose structures were confirmed by MS, H-NMR. The synthesis of the two scaffolds was investigated and a route of cyclization of aryl propargyl ether in high temperature was chose. The reaction conditions such as temperature, solvent, catalysts were optimized. The toxic phosgene was replaced by triphosgene in the synthesis of the ureas. The new process was safe to handle, the yield was improved. These compounds were screened the TNF-(?) inhibiting activity in LPS-induced THP-1 cells, BIRB-796 as reference agonist. The results of in vitro assay indicated that 44 compounds had good inhibiting activity. Among them, the inhibition ratio of 24 compounds was more than 50% when the screening concentration was 3(?)M. 25 compounds were chosen to detect the IC. The result showed that the IC of NEW-21, NEW-22, NEW-25, NEW-27, NEW-35, NEW-36, NEW-42, NEW-48, NEW-56, NEW-57, NEW-59, NEW-62 was lass than 1 (?) M. Among them, NEW-36, NEW-48, NEW-59 exhibited inhibiting activity comparable to that of BIRB-796．The structure-activity relationship of target compounds was investigated according to the biological screening.