Novel hybrids of pyridazine-pyrazoline were synthesized aiming to develop new antiproliferative candidates. All compounds were submitted to the National Cancer Institute (NCI), USA, and many were proved to have significant antiproliferative activity. In addition, in vitro studies of the epidermal growth factor receptor (EGFR) inhibition showed that compounds IXn, IXg, IXb and IXl exhibited excellent inhibitory effect (IC₅₀=0.65, 0.75, 0.82 and 0.84 μM, respectively) compared to Erlotinib (IC₅₀=0.95 μM). The mechanistic effectiveness in cell cycle progression, apoptotic induction and gene regulation were assessed for the promising compounds IXg and IXn due to their significant EGFR inhibition. Flow cytometeric analysis indicated that compounds IXg and IXn result in increased cell numbers in phase G2/M, suggesting cell cycle arrest in phase G2/M in UO-31cells. Furthermore, real time PCR assay illustrated that compounds IXg and IXn elevated Bax/Bcl2 ratio which confirmed the mechanistic pathway of them. Moreover, the apoptotic induction of UO-31 renal cancer cells was enhanced effectively through activation of caspase-3 by compounds IXg and IXn. On the other hand, molecular docking study was performed to investigate binding mode of interaction of compounds with EGFR-PK in the active site with the aim of rationalizing its promising inhibitory activity. Finally, based on the aforementioned findings, compounds IXg and IXn could be considered as effective apoptosis modulators and promising leads for future development of new anti-renal cancer agents.

合成了哒嗪-吡唑啉的新型杂化物，旨在开发新的抗增殖候选物。所有化合物都提交给美国国家癌症研究所 (NCI)，其中许多化合物被证明具有显着的抗增殖活性。此外，在体外的表皮生长因子受体的研究（EGFR）抑制表明化合物IXn，IXG，IXb的和IXL显示出优异的抑制效果（IC ₅₀相比，埃罗替尼（IC = 0.65，0.75，0.82和0.84μM，分别地）₅₀ = 0.95 μM）。评估了有希望的化合物IXg在细胞周期进程、凋亡诱导和基因调控方面的机制有效性和IXn由于它们显着的 EGFR 抑制作用。流式细胞术分析表明，化合物IXg和IXn导致 G2/M 期细胞数量增加，表明 UO-31 细胞中 G2/M 期的细胞周期停滞。此外，实时 PCR 测定表明化合物IXg和IXn提高了 Bax/Bcl2 比率，这证实了它们的机制途径。此外，化合物IXg和IXn通过激活caspase-3有效增强了UO-31肾癌细胞的凋亡诱导. 另一方面，进行分子对接研究以研究化合物与活性位点中 EGFR-PK 相互作用的结合模式，目的是合理化其有希望的抑制活性。最后，基于上述发现，化合物IXg和IXn可被视为有效的细胞凋亡调节剂，并有望为未来开发新的抗肾癌药物提供线索。