Design, synthesis, biological evaluation and molecular modeling of novel 1 H -pyrazolo[3,4-d]pyrimidine derivatives as BRAF V600E and VEGFR-2 dual inhibitors

Abstract Aiming to explore novel BRAF V600E and VEGFR-2 dual inhibitors, a series of 1 H -pyrazolo[3,4-d]pyrimidine derivatives were designed, synthesized and biologically evaluated in this study. Most of the synthesized 1 H -pyrazolo[3,4-d]pyrimidine compounds displayed moderate to high potent activity in both enzymatic and cellular proliferation assays. Among these compounds, 9e , 9g , 9m and 9u showed remarkably high inhibitory activities against both BRAF V600E and VEGFR-2 kinase comparable to positive control Sorafenib. Particularly, compound 9u also showed potent anti-proliferative activity against BRAF V600E -expressing A375 (IC 50  = 1.74 μM) and H-29 (IC 50  = 6.92 μM) as well as VEGFR-2-expressing HUVEC (IC 50  = 5.89 μM), which was also comparable to Sorafenib. Furthermore, kinase selectivity profile showed that 9u had almost poor or no significant inhibitory activity against wild-type BRAF and 15 other tested protein kinases. Flow cytometric analysis showed that compound 9u mainly arrested the A375 and HUVEC cell lines in the G 0 /G 1 stage with a concentration-dependent effect. In addition, the molecular docking and molecular dynamics simulations suggested that 9u adopted a similar binding pattern with Sorafenib at the ATP-binding sites of BRAF V600E and VEGFR-2. Taken together, these results indicated that compound 9u may serve as novel lead compound in research on more effective BRAF V600E and VEGFR-2 dual inhibitors.