Design, synthesis, in vitro anticancer, molecular docking and SAR studies of new series of pyrrolo[2,3-d]pyrimidine derivatives

Abstract

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.

Synthesis, anticancer evaluation of novel hybrid pyrazole-based chalcones, molecular docking, DNA fragmentation, and gene expression: in vitro studies

“…Chalcones have been shown in earlier investigations to induce apoptosis by upregulating the expressions of Bcl-2 and p53 in several cancer cell lines. 2,18,30 …”

“…The growth of cancer is characterized by its rapidity and adaptability, which makes it difficult to find new therapies that will offer more potent therapeutic treatments due to drug resistance and side effects. In light of these results, and in keeping with our research interest in the synthesis of bioactive heterocycles, 2,16–23 we were motivated to create a novel series of chalcones (7a–d and 9a–f) associated with heterocyclic moieties and assess their anti-tumor efficacy in vitro using different human cancer cell lines. The novel chalcones 7d and 9e proved potent and interesting cytotoxic effects against breast cancer cell lines (MCF7) and pancreatic cancer cell lines (PACA2), respectively compared with doxorubicin as a reference drug.…”

Novel 3-(pyrazol-4-yl)-2-(1H-indole-3-carbonyl)acrylonitrile derivatives induce intrinsic and extrinsic apoptotic death mediated P53 in HCT116 colon carcinoma

“…From the readings of the standards, a standard curve was plotted using curve fitting software. Then, the concentrations for unknown samples and controls were measured from the standard curve 87 .…”

“…The DNA gel electrophoresis laddering assay in the HCT116 cell line was performed according to the instructions of Yawata 87 with some modifications. Briefly, after 48 h of exposure of the HCT116 cancer cell line to compound 7f in Petri dishes (60 × 15 mm, Greiner), the cells were suspended in 1 ml of medium and then centrifuged at 900 rpm for 10 min.…”

“…Briefly, after 48 h of exposure of the HCT116 cancer cell line to compound 7f in Petri dishes (60 × 15 mm, Greiner), the cells were suspended in 1 ml of medium and then centrifuged at 900 rpm for 10 min. The genomic DNA was extracted as demonstrated in Yawata 87 . About 1 × 10 6 cells of HCT116 were treated with the IC 50 value of compound 7f .…”