The starting compound 3-amino-1,7-dihydro-4H-pyrazolo[4,3-c]pyridine-4,6(5H)-dione (1) is reacted with each of diketone and β-ketoester, forming pyridopyrazolo[1,5-a]pyrimidines 4a,b and 14a,b, respectively. The compounds 4 and 14 reacted with each of aromatic aldehyde and arenediazonium salt to give the respective arylidenes and arylhydrazo derivatives, respectively. The structure of the new products was established using spectroscopic techniques. The cytotoxic activity of selected targets was tested in vitro against three cancer cell lines MCF7, HepG2 and HCT116. The data obtained from enzymatic assays of TrKA indicated that compounds 7b and 16c have the strongest inhibitory effects on TrKA with IC50 = 0.064 ± 0.0037 μg/ml and IC50 = 0.047 ± 0.0027 μg/ml, respectively, compared to the standard drug Larotrectinib with IC50 = 0.034 ± 0.0021 μg/ml for the HepG2 cancer cell line. In cell cycle analysis, compounds 7b, 15b, 16a and 16c caused the greatest arrest in cell cycle at the G2/M phase. In addition, compound 15b has a higher apoptosis-inducing effect (36.72%) than compounds 7b (34.70%), 16a (21.14) and 16c (26.54%). Compounds 7b, 16a and 16c were shown fit tightly into the active site of the TrKA kinase crystal structure (PDB: 5H3Q). Also, ADME study was performed on some selected potent anticancer compounds described in this study.

中文翻译：

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新型吡啶并[4ʹ,3ʹ:3,4]吡唑并[1,5-a]嘧啶作为潜在原肌球蛋白受体激酶A（TrKA）抑制剂的合成、抗癌评价、分子对接和ADME研究

起始化合物3-氨基-1,7-二氢-4H-吡唑并[4,3-c]吡啶-4,6(5H)-二酮(1)与二酮和β-酮​​酯反应，形成吡啶并吡唑[4,3-c]吡啶-4,6(5H)-二酮(1)。 1,5-a]嘧啶分别为4a、b和14a、b。化合物4和14分别与芳族醛和芳烃重氮盐反应，分别得到亚芳基和芳基亚肼衍生物。使用光谱技术建立了新产品的结构。在体外测试了所选靶标针对三种癌细胞系 MCF7、HepG2 和 HCT116 的细胞毒活性。 TrKA酶法检测数据表明，与IC50标准药物Larotrectinib相比，化合物7b和16c对TrKA具有最强的抑制作用，分别为IC50 = 0.064 ± 0.0037 μg/ml和IC50 = 0.047 ± 0.0027 μg/ml HepG2 癌细胞系 = 0.034 ± 0.0021 μg/ml。在细胞周期分析中，化合物 7b、15b、16a 和 16c 在 G2/M 期引起细胞周期最大程度的停滞。此外，化合物15b比化合物7b(34.70%)、16a(21.14)和16c(26.54%)具有更高的凋亡诱导作用(36.72%)。化合物 7b、16a 和 16c 与 TrKA 激酶晶体结构 (PDB: 5H3Q) 的活性位点紧密贴合。此外，ADME 研究还针对本研究中描述的一些选定的有效抗癌化合物进行了研究。