Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy (Boys), Al-Azhar University, Cairo 11884, Egypt.
Medicinal Chemistry Department, Faculty of Pharmacy, Port Said University, Port Said, 42511, Egypt.
Bioorg Chem. 2024 Apr;145:107223. doi: 10.1016/j.bioorg.2024.107223. Epub 2024 Feb 17.
Herein, we envisioned the design and synthesis of novel pyrazolopyrimidines (confirmed by elemental analysis, H and C NMR, and mass spectra) as multitarget-directed drug candidates acting as EGFR/TOPO II inhibitors, DNA intercalators, and apoptosis inducers. The target diphenyl-tethered pyrazolopyrimidines were synthesized starting from the reaction of phenyl hydrazine and ethoxymethylenemalononitrile to give aminopyrazole-carbonitrile 2. The latter hydrolysis with NaOH and subsequent reaction with 4-chlorobenzaldhyde afforded the corresponding pyrazolo[3,4-d]pyrimidin-4-ol 4. Chlorination of 4 with POCl and sequential reaction with different amines afforded the target compounds in good yields (up to 73 %). The growth inhibition % of the new derivatives (6a-m) was investigated against different cancer and normal cells and the IC values of the most promising candidates were estimated for HNO97, MDA-MB-468, FaDu, and HeLa cancer cells. The frontier derivatives (6a, 6i, 6k, 6l, and 6m) were pursued for their EGFR inhibitory activity. Compound 6l decreased EGFR protein concentration by a 6.10-fold change, compared to imatinib as a reference standard. On the other side, compounds (6a, 6i, 6k, 6l, and 6m) underwent topoisomerase II (TOPO II) inhibitory assay. In particular, compounds 6a and 6l exhibited ICs of 17.89 and 19.39 μM, respectively, surpassing etoposide with IC of 20.82 μM. Besides, the DNA fragmentation images described the great potential of both candidates 6a and 6l in inducing DNA degradation at lower concentrations compared to etoposide and doxorubicin. Moreover, compound 6l, with the most promising EGFR/TOPO II inhibition and DNA intercalation, was selected for further investigation for its apoptosis induction ability by measuring caspases 3, 7, 8, and 9, Bax, p53, MMP2, MMP9, and BCL-2 proteins. Additionally, molecular docking was used to explain the SAR results based on the differences in the molecular features of the investigated congeners and the target receptors' topology.
在这里，我们设想设计和合成新型吡唑并嘧啶（通过元素分析、H 和 C NMR 以及质谱确认）作为多靶点导向药物候选物，作为 EGFR/TOPO II 抑制剂、DNA 嵌入剂和凋亡诱导剂。以苯肼和乙氧亚甲基丙二腈反应得到氨基吡唑-腈 2 为起始原料，合成了目标二苯键合吡唑并嘧啶。后者用 NaOH 水解，再与 4-氯苯甲醛反应，得到相应的吡唑并[3,4-d]嘧啶-4-醇 4。用 POCl 氯化 4，再与不同的胺反应，以良好的收率（高达 73%）得到目标化合物。新衍生物（6a-m）对不同的癌细胞和正常细胞的生长抑制%进行了研究，并对最有前途的候选物 HNO97、MDA-MB-468、FaDu 和 HeLa 癌细胞的 IC 值进行了估算。对最前线的衍生物（6a、6i、6k、6l 和 6m）进行了 EGFR 抑制活性的研究。与参考标准伊马替尼相比，化合物 6l 使 EGFR 蛋白浓度降低了 6.10 倍。另一方面，化合物（6a、6i、6k、6l 和 6m）进行了拓扑异构酶 II（TOPO II）抑制试验。特别是化合物 6a 和 6l 的 IC 分别为 17.89 和 19.39 μM，超过了伊立替康的 IC 20.82 μM。此外，DNA 片段化图像描述了候选物 6a 和 6l 在较低浓度下诱导 DNA 降解的巨大潜力，与依托泊苷和阿霉素相比。此外，化合物 6l 具有最有前途的 EGFR/TOPO II 抑制和 DNA 嵌入作用，选择其进一步研究其诱导细胞凋亡的能力，通过测量半胱天冬酶 3、7、8 和 9、Bax、p53、MMP2、MMP9 和 BCL-2 蛋白。此外，还使用分子对接来解释基于研究同系物和靶受体拓扑结构的分子特征差异的 SAR 结果。
