Annulated pyrazole derivatives as a novel class of urokinase (uPA) inhibitors: Green synthesis, anticancer activity, DNA-damage evaluation, and molecular modelling study

Different series of annulated pyrazole derivatives were designed, synthesized via both green and traditional methods, and structurally characterized. In vitro uPA evaluation, antiproliferative activities and DNA binding damage was studied in this work. Thus, all the synthesized compounds were evaluated against three types of cancer cell lines; HepG-2, HCT-116, and MCF-7 cancer cell lines in addition to normal cell line WI38. Compounds 11, 20, 21, 23 and 24 displayed the most significant antiproliferative activity with IC50 ranging between 4.42 +/- 0.59 mu M to 11.05 +/- 0.95 mu M against HepG-2, HCT-116, and MCF-7 cancer cell lines compared to the reference drug, doxorubicin. Thus compound 11 exhibited cytotoxic activity with IC50 8.58 mu M, 9.22 mu M and 7.53 mu M, compound 20 showed IC50 9.99 mu M, 6.72 mu M and 6.87 mu M, analogue 21 displayed IC50 10.80 mu M, 7.90 mu M and 9.16 mu M, compound 23 showed IC50 4.82 mu M, 11.05 mu M and 4.42 mu M and derivative 24 exhibited potent cytotoxic activity with IC50 7.44 mu M, 5.18 mu M and 8.22 mu M against HepG-2, HCT-116, and MCF-7 cancer cell lines, respectively. Additionally, compounds 11, 21, 23 and 24 showed significant uPA inhibitory activity with IC50 27.28 mu M, 29.36 mu M, 11.73 mu M, and 7.96 mu M respectively. Moreover, HCT-116 cell lines were treated with both compounds 23 and 24 that remarkably showed a high score of DNA binding damage. Mechanistic studies demonstrated the apoptotic activity of the most active tricyclic heteroaromatic analogue 24 on HCT-116 cancer cells by inducing a strong S phase cell cycle arrest suggesting that the mechanism of its antiproliferative activity may be through uPA inhibition. Finally, deeper insight illustrated that the hit compounds exhibited characteristic binding interactions in the active site of uPA that are required in the S pocket, which are important for activity Arg 217, Gly 219, and Ser 190.

Automated summary

Different series of annulated pyrazole derivatives were synthesized and characterized. Compounds 11, 20, 21, 23 and 24 exhibited significant antiproliferative activity against cancer cell lines and showed uPA inhibitory activity and DNA binding damage.