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

摘要

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-氰基苯肼(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的氰基水解为酰氨基得到化合物(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 该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.多个化合物对ALK5信号通路、COX信号通路显示具有很好的抑制活性,并且细胞毒性较小.%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 acyl-chloride(5). Condensation of 6 with 4-cyanophenyl hydrazine(7) gave 2,4,5-triaryl-1H-pyrazol-3(2W)-ones(8). Conversion of the nitrite 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.