The parent 2-(4-benzyl-1-oxophthalazin-2(1H)-yl)-acetohydrazide (4) has twenty-nine compounds. The starting material for their corresponding mono, dipeptides and reactions with active methylene compounds were produced by chemoselective N-alkylation of 4-Benzyl-2H-phthalazin-1-one (2) with ethyl chloroacetate to afford (4-benzyl-1-oxo-1H-phthalazin-2-yl) methyl acetate (3). The ester 3 was hydrazinolyzed to give hydrazide 4, then azide 5 coupled with amino acid ester hydrochloride and/or amines to produce several monopeptides, then the methyl (2-(4-benzyl-1-oxophthalazin-2(1H)-yl) acetyl) glycinate (7a) was hydrazinolyzed to produce corresponding hydrazide 2-(4-benzyl-1-oxophthalazin-2(1H)-yl)-N-(2-hydrazineyl-2-oxo ethyl) acetamide (8a). The hydrazide 8a under azide coupling method was coupled with amino acid ester hydrochloride and/or amines to produce several dipeptides, and the hydrazide 8a was also condensed and/or cyclized with several carbonyl compounds. The cytotoxicity of the synthesized compounds was tested using MTT assay, as well as apoptosis-induction through EGFR inhibition. Compounds 11d, 12c and 12d exhibited potent cytotoxic activities with IC50 values of 0.92, 1.89 and 0.57 μM against MDA-MB-231 cells compared to Erlotinib (IC50 = 1.02 μM). Interestingly compound 12d exhibited promising potent EGFR inhibition with an IC50 value 21.4 nM compared to Erlotinib (IC50 = 80 nM). For apoptosis, compound 12d induced apoptosis in MDA-MB-231 cells by 64.4-fold (42.5% compared to 0.66 for the control); hence, this compound may serve as a potential target-oriented anti-breast cancer agent. These results agreed with the molecular docking studies that highlighted the binding disposition of compound 12d towards EGFR protein. Hence, compound 12d may serve as a potential and selective anti-breast cancer agent.

中文翻译：

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通过 EGFR 介导的细胞凋亡合成二氮杂萘基衍生物作为选择性抗乳腺癌药物：体外和计算机研究


母体2-(4-苄基-1-氧代酞嗪-2(1H)-基)-乙酰肼(4)有二十九种化合物。其相应的单肽、二肽以及与活性亚甲基化合物的反应的起始材料是通过用氯乙酸乙酯对 4-苄基-2H-酞嗪-1-酮 (2) 进行化学选择性 N-烷基化得到 (4-苄基-1-氧代-1H-酞嗪-2-基)乙酸甲酯(3)。酯3经肼解得到酰肼4，然后叠氮化物5与氨基酸酯盐酸盐和/或胺偶联产生几种单肽，然后是甲基(2-(4-苄基-1-氧代酞嗪-2(1H)-基)乙酰基）甘氨酸酯（7a）肼解生成相应的酰肼2-（4-苄基-1-氧代酞嗪-2（1H）-基）-N-（2-肼基-2-氧代乙基）乙酰胺（8a）。叠氮偶联法中的酰肼8a与氨基酸酯盐酸盐和/或胺偶联生成多种二肽，酰肼8a还与多种羰基化合物缩合和/或环化。使用MTT法测试合成化合物的细胞毒性，以及通过EGFR抑制诱导细胞凋亡。与厄洛替尼 (IC50 = 1.02 μM) 相比，化合物 11d、12c 和 12d 对 MDA-MB-231 细胞表现出有效的细胞毒活性，IC50 值为 0.92、1.89 和 0.57 μM。有趣的是，与厄洛替尼（IC50 = 80 nM）相比，化合物 12d 表现出有希望的有效 EGFR 抑制作用，IC50 值为 21.4 nM。对于细胞凋亡，化合物 12d 诱导 MDA-MB-231 细胞凋亡 64.4 倍（42.5%，而对照为 0.66）；因此，该化合物可以作为潜在的靶向抗乳腺癌药物。这些结果与分子对接研究一致，该研究强调了化合物 12d 与 EGFR 蛋白的结合处置。 因此，化合物12d可以作为潜在的、选择性的抗乳腺癌剂。