目前的研究涉及使用微波技术设计和合成新合成的吡咯并[2,3-d]嘧啶衍生物,该衍生物在4和6位含氯原子,在2位含三氯甲基,作为制备此类吡咯并[2,3-d]嘧啶衍生物的新方法。使用光谱和元素分析以及单晶X射线衍射充分表征了合成的吡咯并[2,3-d]嘧啶衍生物3-19的化学结构。所有化合物都在体外针对七种选定的人类癌细胞系进行了测试,即,用MTT法检测MCF7、A549、HCT116、PC3、HePG2、PACA2和BJ1。发现化合物14a,16b和18b对MCF7最活跃,IC50(1.7、5.7和3.4μg/ml,分别)相对于阿霉素(Dox。)(26.1μg/ml)。此外,化合物17对HePG2和PACA2具有有希望的细胞毒性作用,IC50(8.7和6.4μg/ml,分别)相对于Dox。(21.6和28.3μg/ml,分别)。分子对接研究证实了我们的ELISA结果,其显示化合物14a和17对Bcl2抗凋亡蛋白的有希望的结合亲和力。在基因表达水平,P53,BAX,DR4和DR5上调,而Bcl2、Il-8和CDK4在14a中下调,图14b和18b处理MCF7细胞。在蛋白质水平,化合物14b相对于Dox增加了胱天蛋白酶8和BAX(18.263和14.25yg/ml)的活性。(3.99和4.92pg/ml,分别),而与Dox相比,14a处理的MCF7(2.4pg/ml)中Bcl2的活性大大降低。(14.37pg/ml)。化合物14a和14b在MCF7中在G1/S期引起细胞周期停滞。化合物16b和18b诱导MCF7细胞的凋亡性死亡。此外,在14a处理的MCF7细胞中,片段化DNA的百分比显著增加。
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.