当前研究涉及设计和合成一种新合成的吡咯并[2,3-d]嘧啶类衍生物，该衍生物在位置4和6含有氯原子，并在位置2含有三氯甲基基团，使用微波技术作为一种制备该类吡咯并[2,3-d]嘧啶类衍生物的新型坚实方法。合成的吡咯并[2,3-d]嘧啶类衍生物3-19的化学结构通过光谱和元素分析以及单晶X射线衍射得到了良好的表征。所有化合物均通过MTT试验在体外对七种人类癌细胞系进行了检测，分别是MCF7、A549、HCT116、PC3、HePG2、PACA2和BJ1。发现化合物14a、16b和18b对于MCF7具有最活性，其IC50分别为1.7、5.7和3.4μg/ml，而多柔比星(Dox.)的IC50为26.1μg/ml。此外，化合物17对于HePG2和PACA2显示了有希望的细胞毒性作用，其IC50分别为8.7和6.4 μg/ml，相对于多柔比星(Dox.)的IC50分别为21.6和28.3μg/ml。分子对接研究证实了我们的ELISA结果，显示出化合物14a和17与Bcl2抗凋亡蛋白有良好的结合亲和力。在基因表达水平上，P53、BAX、DR4和DR5在14a、14b和18b处理的MCF7细胞中上调，而Bcl2、Il-8和CDK4则下调。在蛋白水平上，相对于多柔比星(Dox.)，化合物14b显著增加了Caspase 8和BAX的活性(分别达到18.263和14.25pg/ml)，而14a处理的MCF7中Bcl2的活性大幅降低(为2.4pg/ml，相对于多柔比星(Dox.)的14.37pg/ml)。化合物14a和14b导致了MCF7细胞在G1/S期的细胞周期停滞。化合物16b和18b引发了MCF7细胞的凋亡死亡。此外，化合物14a处理的MCF7细胞中断裂的DNA百分比显著增加。 © 2023 Springer Nature Switzerland AG.

The current study involves the design and synthesis of a newly synthesized pyrrolo[2,3-d]pyrimidine derivatives to contain chlorine atoms in positions 4 and 6 and trichloromethyl group in position 2 using microwave technique as a new and robust approach for preparation of this type of pyrrolo[2,3-d]pyrimidine derivatives. The chemical structure of the synthesized pyrrolo[2,3-d]pyrimidine derivatives 3-19 was well-characterized using spectral and elemental analyses as well as single-crystal X-ray diffraction. All compounds were tested in vitro against seven selected human cancer cell lines, namely, MCF7, A549, HCT116, PC3, HePG2, PACA2 and BJ1 using MTT assay. It was found that compounds 14a, 16b and 18b were the most active toward MCF7 with IC50 (1.7, 5.7, and 3.4 μg/ml, respectively) relative to doxorubicin (Dox.) (26.1 μg/ml). Additionally, compound 17 exerted promising cytotoxic effects against HePG2 and PACA2 with IC50 (8.7 and 6.4 μg/ml, respectively) relative to Dox. (21.6 and 28.3 μg/ml, respectively). The molecular docking study confirmed our ELISA result which showed the promising binding affinities of compounds 14a and 17 against Bcl2 anti-apoptotic protein. At the gene expression level, P53, BAX, DR4 and DR5 were up-regulated, while Bcl2, Il-8, and CDK4 were down-regulated in 14a, 14b and 18b treated MCF7 cells. At the protein level, compound 14b increased the activity of Caspase 8 and BAX (18.263 and 14.25 pg/ml) relative to Dox. (3.99 and 4.92 pg/ml, respectively), while the activity of Bcl2 was greatly decreased in 14a treated MCF7 (2.4 pg/ml) compared with Dox. (14.37 pg/ml). Compounds 14a and 14b caused cell cycle arrest at the G1/S phase in MCF7. Compounds 16b and 18b induced the apoptotic death of MCF7 cells. In addition, the percentage of fragmented DNA was increased significantly in 14a treated MCF7 cells.© 2023. Springer Nature Switzerland AG.