开发了一种新型高效、通用的取代吡咯并[1,2-a]噻吩并[3,2-e]嘧啶衍生物的一锅三组分合成方法。它基于 2-氨基噻吩和 2-羟基-4-氧代丁-2-烯酸以及二异丙基乙胺催化的氰乙酸衍生物的多步级联反应。结果，在温和的反应条件下以高产率合成了新型吡咯并[1,2-a]噻吩并[3,2-e]嘧啶衍生物（21个化合物）。所开发化合物的结构通过核磁共振和元素分析得到证实。已确定吸电子或供电子取代基对所开发化合物的抗肿瘤活性的影响。对 21 种化合物的体外筛选分析显示，六种主要候选化合物（12aa、12dc、12hc、12ic、12lb 和 12mb）对 B16-F10、4T1 和 CT26 细胞具有最显着的抗肿瘤活性。坏死/凋亡测定表明细胞凋亡是细胞死亡的主要机制。分子对接分析揭示了测试化合物的几个潜在靶点，即磷脂酰肌醇5-磷酸4-激酶（PI5P4K2C）、原癌基因丝氨酸/苏氨酸蛋白激酶（Pim-1）、烟酰胺磷酸核糖转移酶（NAMPT）和二氢叶酸还原酶（DHFR）。先导化合物 (12aa) 可以有效诱导细胞凋亡，具有高产率 (98%)，并且合成时需要低成本的起始化学品。对患有黑色素瘤的小鼠进行的体内实验证实，12aa 化合物在治疗后 15 天即可产生显着的肿瘤抑制作用。特别是，50 mg/kg 剂量组的肿瘤体积为 0.19 cm3，而未治疗小鼠的肿瘤体积为 2.39 cm3，50 mg/kg 组肿瘤重量为 71.6 mg，而未治疗小鼠肿瘤重量为 452.4 mg。因此，我们的结果证明了 12aa 化合物在治疗黑色素瘤方面的巨大潜力，可推荐用于进一步的临床前研究。版权所有 © 2024 Elsevier Inc. 保留所有权利。

A new efficient and versatile one-pot three-component synthesis of substituted pyrrolo[1,2-a]thieno[3,2-e]pyrimidine derivatives has been developed. It is based on a multistep cascade reaction from 2-aminothiophenes and 2-hydroxy-4-oxobut-2-enoic acids, and derivatives of cyanoacetic acid catalyzed by diisopropylethylamine. As a result, novel pyrrolo[1,2-a]thieno[3,2-e]pyrimidine derivatives (21 compounds) were synthesized in a mild reaction conditions with a high yield. The structures of the developed compounds were confirmed by NMR and elemental analysis. The influence of electron-withdrawing or electron-donor substituents on the antitumor activity of the developed compounds has been identified. In vitro screening analysis of 21 compounds revealed six lead candidates (12aa, 12dc, 12hc, 12ic, 12lb, and 12mb) that demonstrated the most significant antitumor activity against B16-F10, 4T1 and CT26 cells. Necrosis/apoptosis assay showed that apoptosis was the predominant mechanism of cell death. Molecular docking analysis revealed several potential targets for tested compounds, i.e. phosphatidylinositol 5-phosphate 4-kinase (PI5P4K2C), proto-oncogene serine/threonine-protein kinase (Pim-1), nicotinamide phosphoribosyltransferase (NAMPT) and dihydrofolate reductase (DHFR). The lead compound (12aa) can effectively induce cell apoptosis, possesses a high yield (98 %) and requires low-cost starting chemicals for its synthesis. In vivo experiments with melanoma-bearing mice confirmed that 12aa compound resulted in the significant tumor inhibition on 15 d after the therapy. In particular, tumor volume was ∼0.19 cm3 for 50 mg/kg versus ∼2.39 cm3 in case of untreated mice and tumor weight was ∼71.6 mg for 50 mg/kg versus ∼452.4 mg when considered untreated mice. Thus, our results demonstrated the high potential of the 12aa compound in the treatment of melanoma and can be recommended for further preclinical studies.Copyright © 2024 Elsevier Inc. All rights reserved.