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磷酸二酯酶4B2(PDE4B2)选择性抑制剂是新型抗炎药物研究的热点。合成异香兰酸芳酰胺衍生物的猪磷酸二酯PDE4强效抑制剂,评价其对人PDE4B2的抑制活性,以明确可用于高通量筛选PDE4抑制剂的靶酶。以异香兰素为原料,经烷基化、氧化、酰胺化反应制得目标化合物;大肠杆菌原核重组表达人PDE4B2;偶联碱性磷酸酶的孔雀绿定磷法测定酶活性,96孔板高通量测定化合物的抑制常数。结果显示,结构确认的5个异香兰酸芳酰胺衍生物纯度超过88%,其对人PDE4B2的抑制常数明显高于对猪主动脉PDE4的抑制常数。不同来源PDE4对异香兰酸芳酰胺衍生物敏感性不同,拟用人PDE4B2为靶酶进行抑制剂高通量筛选。
Phosphodiesterase 4B2 (PDE4B2) selective inhibitor is a new hot research topic of anti-inflammatory drugs. A potent PDX4 inhibitor of porcine phosphodiester derivatives was synthesized and its inhibitory activity against human PDE4B2 was evaluated in order to make it clear that it could be used as a target enzyme for high-throughput screening of PDE4 inhibitors. The target compound was obtained by alkylation, oxidation and amidation reaction of isovanillin. The prokaryotic recombinant human PDE4B2 was expressed in E. coli, and the activity of malachite green phosphorus coupled with alkaline phosphatase Flux The inhibition constant of the compound. The results showed that the structurally confirmed 5-isoprenoid arylamide derivatives were over 88% pure, and their inhibitory constants against human PDE4B2 were significantly higher than those against porcine aortic PDE4. Different sources of PDE4 different sensitivity of different derivatives of aramide aromatic amide, intended to use human PDE4B2 target enzyme inhibitor high-throughput screening.