Development of novel quinazolinone-based compounds with anti-proliferative activity as dual STAT-3 and c-Src inhibitors: design, synthesis and molecular dynamic studies

A series of quinazolinone derivatives 5a, 5b, 6a-f, 7a-c, 8a-e, and 12a-d with different moieties as dual STAT-3/c-Src inhibitors were designed, synthesized, and evaluated for anti-cancer activity. In vitro anti-cancer screening was carried out against 60 human Cancer cell lines by (NCI) for all the synthesized compounds 5a, 5b, 6a-f, 7a-c, 8a-e, and 12a-d. All the new compounds showed anti-cancer inhibitory activity against various human cell lines. Fortunately, the oxime derivative 7c and the 4-oxoquinazoline-acetohydrazide derivative 12d showed the highest potency and the broadest spectrum of activity among all Other analogs against the tested Cancer cell lines. Both compounds 7c and 12d were chosen by NCI for five dose evaluation against 60 human Cancer cell lines. Compounds 7c and 12d showed significant anti-cancer activity against both melanoma (SK-MEL-2) cell line (GI₅₀ = -5.79 and -5.75 μM, respectively) and CNS Cancer (SNB-75) cell line (GI₅₀ = -5.68 and -5.63 μM, respectively). The synthesized derivatives 7c and 12d were evaluated against both STAT-3 and c-Src Enzymes. Surprisingly, both derivatives 7c and 12d demonstrated more potent inhibitory activity (IC₅₀ = 1.291 ± 0.055 and 0.844 ± 0.036 μM, sequentially) towards STAT-3 enzyme compared to the reference drug STAT3-IN-3 (IC₅₀ = 2.1 ± 0.20 μM). Also, the pyrazoloquinazoline derivative 12d (IC₅₀ = 0.268 ± 0.011 μM) showed highest inhibitory activity than the hydroxyimino derivative 7c (IC₅₀ = 0.565 ± 0.024 μM) against c-Src enzyme compared to staurosporin (IC₅₀ = 0.139 ± 0.006 μM) as reference drug. Compounds 7c and 12d exhibited dual STAT-3/c-Src inhibitory activity. Moreover, cell cycle analysis and Apoptosis were performed to compound 12d against both SNB-75 and SK-MEL-2. Compound 12d showed cell cycle arrest at G0/G1 and G2/M phases on SNB-75 cell line and also showed cell cycle arrest at G0/G1 on SK-MEL-2 cell line, Additionally, 12d induced a necrotic percentage of 3.66 % and 3.50 %, respectively towards SK-MEL-2 and SNB-75 cells which were significantly higher than control that showed only 1.58 % and 1.94 % necrosis. ADME study showed that compound 7c demonstrated bioavailability score of 0.55 similar to that of staurosporin. The modeling and molecular dynamic results were performed for the most potent derivatives 7c and 12d. Fourtunately, the dual activity against STAT-3/c-Src of the most potent derivatives 7c and 12d were found to enrich the anti-proliferative activity of them.

Anti-proliferative activity; CNS cancer; Melanoma; Quinazolinones; STAT-3/c-Src.