Synthesis, antitumor evaluation and microarray study of some new pyrazolo[3,4-d][1,2,3]triazine derivatives

Design and synthesis of new anticancer scaffolds; pyrazolo[3,4-d][1,2,3]triazine derivatives, is a promising solution to overcome drug resistance problem. A series of (E)-2-cyano-N-(aryl)-3-methylthio-3-(substituted-amino)acrylamides 3a-e was synthesized and transformed to the 3-aminopyrazole derivatives 4a-e which were then transformed to the target pyrazolotriazinones 6a-e. All compounds were evaluated for their anticancer activity against three different cancer cell lines namely Huh-7, Panc-1 and CCRF. Compounds 3a, 3c, 6a and 6c showed excellent anticancer activity against Huh-7 cell line (IC50: 4.93–8.84 μM vs doxorubicin 5.43 μM). Similarly, compounds 6a and 6d showed excellent activities against Panc-1 cells (IC50: 9.91 μM and 4.93 μM vs doxorubicin 6.90 μM). Caspase-Glo 3/7 assay was done and the results revealed that the pro-apoptotic activity of the target compounds could be due to the stimulation of caspases 3/7. Microarray experiment for Huh-7 cells treated with 6c was performed to search for other molecular changes. SLC26A3, UGT1A1, UGT2B15, UGT2B7, DNASE1, MUCDH1 and UGT2B17 were among the up-regulated genes, while, GIP3, TAGL, THBS1, IFI27, FSCN1 and SOCS2 were among the most extensively down-regulated genes. These genes belong to apoptosis, metabolism, cell cycle, tumor growth and suppressor genes. Finally, pyrazolo[3,4-d][1,2,3]triazine derivatives could be potent anticancer drugs in the future. New pyrazolo[3,4-d][1,2,3]triazine derivatives were synthesized and evaluated in vitro for their anticancer activity. Compound 6c showed excellent anticancer activity. In this study we provide new anticancer scaffolds to overcome drug resistance problem. [Display omitted] •New pyrazolo[3,4-d][1,2,3]triazine derivatives were synthesized.•Pyrazolotriazines showed excellent antitumor activities against Huh-7 and Panc-1 cell lines.•Pyrazolotriazines 6a and 6c activate caspases 3/7.•6c Up-regulates SLC26A3, UGT1A1, UGT2B15, UGT2B7, DNASE1, MUCDH1 and UGT2B17 genes.•Pyrazolotriazine 6c down-regulates GIP3, TAGL, THBS1, IFI27, FSCN1 and SOCS2 genes.