Gümus Mehmet, Ozgur Aykut, Tutar Lutfi, Disli Ali, Koca Irfan, Tutar Yusuf
Department of Chemistry, Faculty of Arts and Sciences, Bozok University, Yozgat, Turkey.
Cumhuriyet University, Faculty of Pharmacy, Basic Pharmaceutical Sciences Department, Division of Biochemistry, 58140, Sivas, Turkey.
Curr Pharm Biotechnol. 2016;17(14):1231-1245. doi: 10.2174/1389201017666161031105815.
Despite development of novel cancer drugs, invasive ductal breast carcinoma and its metastasis are still highly morbid. Therefore, new therapeutic approaches are being developed and Hsp90 is an important target for drug design. For this purpose, a series of benzodiazepine derivatives were designed and synthesized as novel Hsp90 inhibitor.
Benzodiazepine derivatives anticancer activities were determined by XTT cell proliferation assay against human breast cancer cell line (MCF-7). Effects of the compounds on endothelial function were monitored on human vascular endothelium (HUVEC) cell line as well. In order to determine the anti-proliferative mechanism of the compounds, in silico molecular docking studies were performed between Hsp90 ATPase domain and the benzodiazepine derivatives. Further, these compounds perturbation on Hsp90 ATPase function were tested. Fluorescence binding experiments showed that the derivatives bind Hsp90 effectively. Expression analysis of known cancer drug target genes by PCR array experiments suggest that the benzodiazepine derivatives have remarkable anticancer activity.
A representative Benzodiazepine derivative D5 binds Hsp90 with Kd value of 3,93 μM and with estimated free energy of binding -7.99 (kcal/mol). The compound decreases Hsp90 ATPase function and inhibit Hsp90 client protein folding activity. The compound inhibits expression of both Hsp90 isoforms and key proteins (cell cycle receptors; PLK2 and TERT, kinases; PI3KC3 and PRKCE, and growth factors; IGF1, IGF2, KDR, and PDGFRA) on oncogenic pathways.
Benzodiazepine derivatives presented here display anticancer activity. The compounds effect on both breast cancer and endothelial cell lines show their potential as drug templates to inhibit breast cancer and its metastasis.
尽管新型抗癌药物不断研发，但浸润性导管乳腺癌及其转移仍具有高致死率。因此，新的治疗方法正在不断开发，而热休克蛋白90（Hsp90）是药物设计的重要靶点。为此，设计并合成了一系列苯二氮䓬衍生物作为新型Hsp90抑制剂。
通过XTT细胞增殖试验测定苯二氮䓬衍生物对人乳腺癌细胞系（MCF-7）的抗癌活性。同时也在人血管内皮（HUVEC）细胞系上监测这些化合物对内皮功能的影响。为了确定化合物的抗增殖机制，对Hsp90 ATP酶结构域与苯二氮䓬衍生物进行了计算机模拟分子对接研究。此外，还测试了这些化合物对Hsp90 ATP酶功能的干扰。荧光结合实验表明，这些衍生物能有效结合Hsp90。通过PCR阵列实验对已知癌症药物靶基因进行表达分析，结果表明苯二氮䓬衍生物具有显著的抗癌活性。
一种代表性的苯二氮䓬衍生物D5与Hsp90结合，解离常数（Kd）值为3.93 μM，结合自由能估计为-7.99（千卡/摩尔）。该化合物降低Hsp90 ATP酶功能，并抑制Hsp90客户蛋白的折叠活性。该化合物抑制Hsp90两种亚型以及致癌途径上关键蛋白（细胞周期受体；PLK2和TERT，激酶；PI3KC3和PRKCE，以及生长因子；IGF1、IGF2、KDR和PDGFRA）的表达。
本文介绍的苯二氮䓬衍生物具有抗癌活性。这些化合物对乳腺癌细胞系和内皮细胞系均有作用，显示出它们作为抑制乳腺癌及其转移的药物模板的潜力。
