新型杂环亚胺连接香豆素-噻唑杂合体的合成及生物评价作为抗癌剂。
Synthesis and Biological Evaluation of Novel Heterocyclic Imines Linked Coumarin- Thiazole Hybrids as Anticancer Agents.
机构信息
Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research (NIPER), Hyderabad 500 037, India.
Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research (NIPER), Hyderabad 500 037, India.
出版信息
Anticancer Agents Med Chem. 2019;19(4):557-566. doi: 10.2174/1871520619666190207140120.
BACKGROUND
Human Galectin-1, a protein of lectin family showing affinity towards β-galactosides has emerged as a critical regulator of tumor progression and metastasis, by modulating diverse biological events including homotypic cell aggregation, migration, apoptosis, angiogenesis and immune escape. Therefore, galectin-1 inhibitors might represent novel therapeutic agents for cancer.
METHODS
A new series of heterocyclic imines linked coumarin-thiazole hybrids (6a-6r) was synthesized and evaluated for its cytotoxic potential against a panel of six human cancer cell lines namely, lung (A549), prostate (DU-145), breast (MCF-7 & MDA-MB-231), colon (HCT-15 & HT-29) using MTT assay. Characteristic apoptotic assays like DAPI staining, cell cycle, annexin V and Mitochondrial membrane potential studies were performed for the most active compound. Furthermore, Gal-1 inhibition was confirmed by ELISA and fluorescence spectroscopy.
RESULTS
Among all, compound 6g {3-(2-(2-(pyridin-2-ylmethylene) hydrazineyl) thiazol-4-yl)-2H-chromen-2- one} exhibited promising growth inhibition against HCT-15 colorectal cancer cells with an IC50 value of 1.28 ± 0.14 µM. The characteristic apoptotic morphological features like chromatin condensation, membrane blebbing and apoptotic body formation were clearly observed with compound 6g on HCT-15 cells using DAPI staining studies. Further, annexin V-FITC/PI assay confirmed effective early apoptosis induction by treatment with compound 6g. Loss of mitochondrial membrane potential and enhanced ROS generation were confirmed with JC-1 and DCFDA staining method, respectively by treatment with compound 6g, suggesting a possible mechanism for inducing apoptosis. Moreover, flow cytometric analysis revealed that compound 6g blocked G0/G1 phase of the cell cycle in a dose-dependent manner. Compound 6g effectively reduced the levels of Gal-1 protein in a dose-dependent manner. The binding constant (Ka) of 6g with Gal-1 was calculated from the intercept value which was observed as 1.9 x 107 M-1 by Fluorescence spectroscopy. Molecular docking studies showed strong interactions of compound 6g with Gal-1 protein.
CONCLUSION
Our studies demonstrate the anticancer potential and Gal-1 inhibition of heterocyclic imines linked coumarin-thiazole hybrids.
背景
半乳糖凝集素-1（Galectin-1）是凝集素家族中的一种蛋白，对β-半乳糖苷具有亲和力，它通过调节包括同质细胞聚集、迁移、凋亡、血管生成和免疫逃逸在内的多种生物学事件，成为肿瘤进展和转移的关键调节剂。因此，Galectin-1 抑制剂可能成为癌症的新型治疗药物。
方法
合成了一系列新型杂环亚胺连接的香豆素-噻唑杂合体（6a-6r），并通过 MTT 法评估其对六种人癌细胞系（肺（A549）、前列腺（DU-145）、乳腺（MCF-7 和 MDA-MB-231）、结肠（HCT-15 和 HT-29））的细胞毒性潜力。对最活跃的化合物进行了特征性的凋亡检测，如 DAPI 染色、细胞周期、膜联蛋白 V 和线粒体膜电位研究。此外，通过 ELISA 和荧光光谱法证实了 Gal-1 的抑制作用。
结果
在所研究的化合物中，化合物 6g（3-（2-（2-（吡啶-2-基亚甲基）肼基）噻唑-4-基）-2H-色烯-2-酮）对 HCT-15 结肠癌细胞的生长抑制作用最为显著，IC50 值为 1.28 ± 0.14 µM。通过 DAPI 染色研究，在 HCT-15 细胞上用化合物 6g 处理后，可观察到染色质浓缩、细胞膜起泡和凋亡小体形成等典型的凋亡形态特征。此外，用膜联蛋白 V-FITC/PI 检测证实了化合物 6g 有效诱导早期凋亡。用 JC-1 和 DCFDA 染色法分别证实了线粒体膜电位丧失和 ROS 生成增加，这表明了诱导凋亡的可能机制。此外，流式细胞术分析显示，化合物 6g 以剂量依赖的方式阻断细胞周期的 G0/G1 期。化合物 6g 能有效降低 Gal-1 蛋白的水平，且呈剂量依赖性。荧光光谱法的截距值表明化合物 6g 与 Gal-1 的结合常数（Ka）为 1.9 x 107 M-1。分子对接研究表明，化合物 6g 与 Gal-1 蛋白有很强的相互作用。
结论
本研究证明了杂环亚胺连接的香豆素-噻唑杂合体具有抗癌潜力和 Gal-1 抑制作用。
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