2,4,5-三芳基-1H-吡唑-3(2H)-酮类化合物的合成与生物活性评价

目的 设计合成2,4,5-三芳基-1H-吡唑-3(2H)-酮类化合物，并研究其对ALK5信号通路、COX-1和COX-2信号通路的抑制活性，以期发现新型的ALK5或COX抑制剂。方法 关键中间体3-氧代-2,3-芳基丙酸甲酯（6）可以由两种方法制备：一是由芳基醛（1）与芳基乙酸甲酯（2）Aldol缩合后经Swern氧化的方法得到；二是通过芳基酰氯（5）与芳基乙酸甲酯的钠盐（4）直接缩合得到。化合物6与4-腈基苯肼（7）缩合得到4-（3，4-二芳基-5-氧代吡唑啉-1-基]）苯腈（8），将化合物8的腈基水解为酰氨得到化合物（9）。应用基于细胞的TGF-Smad2检测评价化合物的ALK5抑制活性; 采用化学发光法测试化合物对COX1和COX2的抑制活性；采用MTT法检测化合物的细胞毒性。结果与结论 本文所合成的化合物和中间体均为新化合物，所有目标化合物和大部分中间体的结构经过了核磁与质谱的确证，其中目标化合物18个。多个化合物显示具有很好的对ALK5信号通路、COX信号通路的抑制活性，并且细胞毒性较小。

Synthesis and biological evaluation of 2,4,5-triaryl- 1H-pyrazol-3(2H)-ones

To explore novel ALK5 inhibitors or COX inhibitors,eighteen novel 2,4,5-triaryl-1H-pyrazol-3(2H)-one derivatives were designed and synthesized,and their inhibitory activities against ALK5 and COX in vitro along with the preliminary SAR were investigated.The key intermediates methyl 3-keto-2,3-diaryl-propionates(6) were prepared by Aldol condensation of aryl aldehyde(1) with methyl aryl acetate(2) followed by Swern oxidation or through acylation of the sodium enolate of methyl aryl acetate(4) with acylchloride(5).Condensation of 6 with 4-cyanophenyl hydrazine(7) gave 2,4,5-triaryl-1H-pyrazol-3(2H)-ones(8).Conversion of the nitrile group of 8 to carboxamide afforded compound 9.All target compounds and intermediates are novel compounds.The structure of all target compounds and most intermediates were confirmed by 1H-NMR and MS spectra.All the target compounds were evaluated for their ALK5 inhibitory activity and cytotoxicity in TGF-Smad2 assay and MTT assay respectively,some compounds were evaluated for their COX-1 and COX-2 inhibitory activity in chemiluminescent COX inhibitor screening assay.The results demonstrated that most compounds exhibited noticeable ALK5 inhibition activities at 1 μmol·L-1,while displayed no significant cytotoxicities at 30 μmol·L-1.The most activity compound 8d and 8e showed about 35% ALK5 phosphorylation inhibition at 1 μmol·L-1,which was a little weaker compared with SB-431542.All tested compounds,namely 8b,9b,9c,8d,9d,8e,8f,9f,8g and 9g showed more potent COX-1 inhibitory activities at the concentration of both 3×10-6 mol·L-1 and 1×10-4 mol·L-1 when compared with indometacin.Compounds 8d,8e and 9g exhibited better COX-2 inhibitory activities than celecoxib at concentration of 1×10-7 mol·L-1 and 3×10-6 mol·L-1.Compounds 8f,9f and 8b were more potent at inhibiting COX-2 than COX-1 at 3×10-6 mol·L-1.