Department of Pathology, Kazan State Medical University, 420012 Kazan, Russia.
Department of Radiotherapy and Radiology, Russian Medical Academy of Continuous Professional Education, 125993 Moscow, Russia.
Molecules. 2022 Apr 30;27(9):2873. doi: 10.3390/molecules27092873.
Despite the tubulin-binding agents (TBAs) that are widely used in the clinic for cancer therapy, tumor resistance to TBAs (both inherited and acquired) significantly impairs their effectiveness, thereby decreasing overall survival (OS) and progression-free survival (PFS) rates, especially for the patients with metastatic, recurrent, and unresectable forms of the disease. Therefore, the development of novel effective drugs interfering with the microtubules' dynamic state remains a big challenge in current oncology. We report here about the novel ethyl 2-amino-1-(furan-2-carboxamido)-5-(2-aryl/tert-butyl-2-oxoethylidene)-4-oxo-4,5-dihydro-1H-pyrrole-3-carboxylates (EAPCs) exhibiting potent anti-cancer activities against the breast and lung cancer cell lines in vitro. This was due to their ability to inhibit tubulin polymerization and induce cell cycle arrest in M-phase. As an outcome, the EAPC-treated cancer cells exhibited a significant increase in apoptosis, which was evidenced by the expression of cleaved forms of PARP, caspase-3, and increased numbers of Annexin-V-positive cells. By using the in silico molecular modeling methods (e.g., induced-fit docking, binding metadynamics, and unbiased molecular dynamics), we found that EAPC-67 and -70 preferentially bind to the colchicine-binding site of tubulin. Lastly, we have shown that the EAPCs indicated above and colchicine utilizes a similar molecular mechanism to inhibit tubulin polymerization via targeting the T7 loop in the β-chain of tubulin, thereby preventing the conformational changes in the tubulin dimers required for their polymerization. Collectively, we identified the novel and potent TBAs that bind to the colchicine-binding site and disrupt the microtubule network. As a result of these events, the compounds induced a robust cell cycle arrest in M-phase and exhibited potent pro-apoptotic activities against the epithelial cancer cell lines in vitro.
尽管临床上广泛使用微管结合剂（TBAs）治疗癌症，但肿瘤对 TBAs 的耐药性（包括遗传性和获得性）显著降低了其疗效，从而降低了总生存率（OS）和无进展生存率（PFS），尤其是对于转移性、复发性和不可切除的疾病患者。因此，开发新型有效的药物来干扰微管的动态状态仍然是当前肿瘤学的一个重大挑战。我们在这里报告了新型的乙基 2-氨基-1-（呋喃-2-甲酰胺基）-5-（2-芳基/叔丁基-2-氧代乙基）-4-氧代-4,5-二氢-1H-吡咯-3-羧酸酯（EAPCs），它们在体外对乳腺癌和肺癌细胞系表现出很强的抗癌活性。这是由于它们抑制微管聚合和诱导细胞周期停滞在 M 期的能力。结果，EAPC 处理的癌细胞表现出明显的凋亡增加，这表现为 PARP、caspase-3 的裂解形式的表达增加和 Annexin-V 阳性细胞数量的增加。通过使用计算机分子建模方法（例如，诱导拟合对接、结合代谢动力学和无偏分子动力学），我们发现 EAPC-67 和-70 优先结合到微管的秋水仙碱结合位点。最后，我们表明，上述 EAPCs 和秋水仙碱利用相似的分子机制通过靶向微管β链的 T7 环来抑制微管聚合，从而阻止微管二聚体聚合所需的构象变化。总之，我们确定了新型有效的 TBAs，它们结合到秋水仙碱结合位点并破坏微管网络。由于这些事件，这些化合物在 M 期引起了强烈的细胞周期停滞，并在体外对上皮癌细胞系表现出很强的促凋亡活性。
