Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Biruni University, 34010, Zeytinburnu, Istanbul, Turkey.
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Istanbul Medipol University, 34815 Beykoz-Istanbul, Turkey.
Curr Med Chem. 2024;31(24):3798-3817. doi: 10.2174/0929867330666230626143911.
Recent studies showed that the cooperation between c-SRC and EGFR is responsible for more aggressive phenotype in diverse tumors, including glioblastomas and carcinomas of the colon, breast, and lung. Studies show that combination of SRC and EGFR inhibitors can induce apoptosis and delay the acquired resistance to chemotherapy. Therefore, such combination may lead to a new therapeutic strategy for the treatment of EGFR-mutant lung cancer. Osimertinib was developed as a third-generation EGFR-TKI to combat the toxicity of EGFR mutant inhibitors. Due to the resistance and adverse reaction of osimertinib and other kinase inhibitors, 12 novel compounds structurally similar to osimertinib were designed and synthesized.
Compounds were synthesized by developing novel original synthesis methods and receptor interactions were evaluated through a molecular docking study. To evaluate their inhibitory activities against EGFR and SRC kinase, enzyme assays were used. Anticancer potencies were determined using lung, breast, prostate (A549, MCF6, PC3) cancer cell lines. Compounds were also tested against normal (HEK293) cell line to evaluate their cyctotoxic effects.
Although, none of compounds showed stronger inhibition compared to osimertinib in the EGFR enzyme inhibition studies, compound 16 showed the highest efficacy with an IC of 1.026 μM. It also presented potent activity against SRC kinase with an IC of 0.002 μM. Among the tested compounds, the urea containing derivatives 6-11 exhibited a strong inhibition profile (80.12-89.68%) against SRC kinase in comparison to the reference compound dasatinib (93.26%). Most of the compounds caused more than 50% of cell death in breast, lung and prostate cancer cell lines and weak toxicity for normal cells in comparison to reference compounds osimertinib, dasatinib and cisplatin. Compound 16 showed strong cytotoxicity on lung and prostate cancer cells. Treatment of prostate cancer cell lines with the most active compound, 16, significantly increased the caspase-3 (8-fold), caspase-8 (6-fold) and Bax (5.7-fold) levels and decreased the Bcl-2 level (2.3-fold) compared to the control group. These findings revealed that the compound 16 strongly induces apoptosis in the prostate cancer cell lines.
Overall kinase inhibition, cytotoxicity and apoptosis assays presented that compound 16 has dual inhibitory activity against SRC and EGFR kinases while maintaining low toxicity against normal cells. Other compounds also showed considerable activity profiles in kinase and cell culture assays.
最近的研究表明，c-SRC 与 EGFR 的合作是导致多种肿瘤（包括神经胶质瘤和结肠、乳腺和肺的癌）更具侵袭性表型的原因。研究表明，SRC 和 EGFR 抑制剂的联合使用可以诱导细胞凋亡并延缓对化疗的获得性耐药。因此，这种联合治疗可能为治疗 EGFR 突变型肺癌提供新的治疗策略。奥希替尼是一种第三代 EGFR-TKI，旨在对抗 EGFR 突变抑制剂的毒性。由于奥希替尼和其他激酶抑制剂的耐药性和不良反应，设计并合成了 12 种结构与奥希替尼相似的新型化合物。
通过开发新的原创合成方法来合成化合物，并通过分子对接研究评估受体相互作用。为了评估它们对 EGFR 和 SRC 激酶的抑制活性，使用酶测定法进行了检测。使用肺癌、乳腺癌、前列腺癌（A549、MCF6、PC3）癌细胞系来确定抗癌潜力。还测试了化合物对正常（HEK293）细胞系的细胞毒性作用。
虽然在 EGFR 酶抑制研究中，没有一种化合物的抑制作用强于奥希替尼，但化合物 16 的疗效最高，IC 为 1.026 μM。它对 SRC 激酶也具有很强的活性，IC 为 0.002 μM。在所测试的化合物中，与参比化合物 dasatinib（93.26%）相比，含脲的衍生物 6-11 对 SRC 激酶表现出强烈的抑制作用（80.12-89.68%）。与参比化合物奥希替尼、dasatinib 和顺铂相比，大多数化合物在乳腺癌、肺癌和前列腺癌细胞系中引起超过 50%的细胞死亡，对正常细胞的毒性较弱。化合物 16 对肺癌和前列腺癌细胞表现出很强的细胞毒性。用最活跃的化合物 16 处理前列腺癌细胞系，与对照组相比，caspase-3（8 倍）、caspase-8（6 倍）和 Bax（5.7 倍）水平显著增加，Bcl-2 水平（2.3 倍）降低。这些发现表明，化合物 16 可强烈诱导前列腺癌细胞系凋亡。
总体激酶抑制、细胞毒性和细胞凋亡检测结果表明，化合物 16 对 SRC 和 EGFR 激酶具有双重抑制活性，同时对正常细胞保持低毒性。其他化合物在激酶和细胞培养测定中也表现出相当大的活性谱。
