Department of Pharmaceutical Sciences, College of Pharmacy (K.E.A., D.-J.H., D.M., D.D.M., W.L.), and Department of Pathology (D.N.P., T.N.S.), the University of Tennessee Health Science Center, Memphis, Tennessee; State Key Laboratory of Biotherapy and Cancer Center, Collaborative Innovation Center of Biotherapy (Y.W., Q.C., J.Y.), and Department of Respiratory Medicine (Y.W.), West China Hospital, Sichuan University, Chengdu, China; Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, New York (Z.-N.L., Z.-S.C.); and Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, Tennessee (G.K., S.W.W.).
Department of Pharmaceutical Sciences, College of Pharmacy (K.E.A., D.-J.H., D.M., D.D.M., W.L.), and Department of Pathology (D.N.P., T.N.S.), the University of Tennessee Health Science Center, Memphis, Tennessee; State Key Laboratory of Biotherapy and Cancer Center, Collaborative Innovation Center of Biotherapy (Y.W., Q.C., J.Y.), and Department of Respiratory Medicine (Y.W.), West China Hospital, Sichuan University, Chengdu, China; Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, New York (Z.-N.L., Z.-S.C.); and Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, Tennessee (G.K., S.W.W.)
Mol Pharmacol. 2019 Jul;96(1):73-89. doi: 10.1124/mol.118.114801. Epub 2019 May 1.
Interfering with microtubule dynamics is a well-established strategy in cancer treatment; however, many microtubule-targeting agents are associated with drug resistance and adverse effects. Substantial evidence points to ATP-binding cassette (ABC) transporters as critical players in the development of resistance. Herein, we demonstrate the efficacy of DJ95 (2-(1-indol-6-yl)-4-(3,4,5-trimethoxyphenyl)-1-imidazo[4,5-]pyridine), a novel tubulin inhibitor, in a variety of cancer cell lines, including malignant melanomas, drug-selected resistant cell lines, specific ABC transporter-overexpressing cell lines, and the National Cancer Institute 60 cell line panel. DJ95 treatment inhibited cancer cell migration, caused morphologic changes to the microtubule network foundation, and severely disrupted mitotic spindle formation of mitotic cells. The high-resolution crystal structure of DJ95 in complex with tubulin protein and the detailed molecular interactions confirmed its direct binding to the colchicine site. In vitro pharmacological screening of DJ95 using SafetyScreen44 (Eurofins Cerep-Panlabs) revealed no significant off-target interactions, and pharmacokinetic analysis showed that DJ95 was maintained at therapeutically relevant plasma concentrations for up to 24 hours in mice. In an A375 xenograft model in nude mice, DJ95 inhibited tumor growth and disrupted tumor vasculature in xenograft tumors. These results demonstrate that DJ95 is potent against a variety of cell lines, demonstrated greater potency to ABC transporter-overexpressing cell lines than existing tubulin inhibitors, directly targets the colchicine binding domain, exhibits significant antitumor efficacy, and demonstrates vascular-disrupting properties. Collectively, these data suggest that DJ95 has great potential as a cancer therapeutic, particularly for multidrug resistance phenotypes, and warrants further development. SIGNIFICANCE STATEMENT: Paclitaxel is a widely used tubulin inhibitor for cancer therapy, but its clinical efficacy is often limited by the development of multidrug resistance. In this study, we reported the preclinical characterization of a new tubulin inhibitor DJ95, and demonstrated its abilities to overcome paclitaxel resistance, disrupt tumor vasculature, and exhibit significant antitumor efficacy.
干扰微管动力学是癌症治疗中一种成熟的策略；然而，许多微管靶向药物与耐药性和不良反应相关。大量证据表明，ATP 结合盒（ABC）转运蛋白是耐药性发展的关键因素。在此，我们证明了新型微管抑制剂 DJ95（2-(1-吲哚-6-基)-4-(3,4,5-三甲氧基苯基)-1-咪唑并[4,5-]吡啶）在多种癌细胞系中的疗效，包括恶性黑色素瘤、药物选择的耐药细胞系、特定 ABC 转运体过表达细胞系和国家癌症研究所 60 细胞系面板。DJ95 处理抑制癌细胞迁移，导致微管网络基础的形态变化，并严重破坏有丝分裂细胞的有丝分裂纺锤体形成。DJ95 与微管蛋白复合物的高分辨率晶体结构和详细的分子相互作用证实了其与秋水仙碱结合位点的直接结合。使用 SafetyScreen44（Eurofins Cerep-Panlabs）对 DJ95 进行体外药理学筛选显示没有明显的脱靶相互作用，药代动力学分析表明，DJ95 在小鼠体内可维持治疗相关的血浆浓度长达 24 小时。在裸鼠 A375 异种移植模型中，DJ95 抑制肿瘤生长并破坏异种移植肿瘤中的肿瘤血管。这些结果表明，DJ95 对多种细胞系有效，对 ABC 转运体过表达细胞系的活性大于现有的微管抑制剂，直接靶向秋水仙碱结合域，具有显著的抗肿瘤疗效，并具有血管破坏特性。总之，这些数据表明，DJ95 作为一种癌症治疗药物具有很大的潜力，特别是对于多药耐药表型，值得进一步开发。意义：紫杉醇是一种广泛用于癌症治疗的微管抑制剂，但它的临床疗效往往受到多药耐药性的限制。在这项研究中，我们报告了新型微管抑制剂 DJ95 的临床前特征，并证明了它克服紫杉醇耐药性、破坏肿瘤血管和表现出显著抗肿瘤疗效的能力。
