A novel small-molecule inhibitor of mcl-1 blocks pancreatic cancer growth in vitro and in vivo.
A novel small-molecule inhibitor of mcl-1 blocks pancreatic cancer growth in vitro and in vivo.
复制标题
MCL-1的一种新型的小分子抑制剂阻断了体外和体内胰腺癌的生长。
DOI：
10.1158/1535-7163.mct-12-0767
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发表时间：
2014-03
影响因子：
5.7
通讯作者：
Nikolovska-Coleska Z
中科院分区：
文献类型：
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作者：
Abulwerdi F;Liao C;Liu M;Azmi AS;Aboukameel A;Mady AS;Gulappa T;Cierpicki T;Owens S;Zhang T;Sun D;Stuckey JA;Mohammad RM;Nikolovska-Coleska Z
使用高通量筛选（HTS）方法，我们已经鉴定并验证了几种小分子Mcl-1抑制剂（SMIs）。在这里，我们描述了一种新的选择性Mcl-1 SMI抑制剂2 (UMI-77)，它是通过对先导化合物1 （UMI-59）进行基于结构的化学修饰而开发的。我们利用互补的生化、生物物理和计算方法表征了UMI-77与Mcl-1的结合，并测定了其对胰腺癌（PC）细胞和体内异种移植模型的抗肿瘤活性。UMI-77与Mcl-1的BH3结合槽结合，Ki值为490 nM，对其他抗凋亡Bcl-2成员具有选择性。UMI-77抑制细胞生长，诱导细胞凋亡，并伴有细胞色素c的释放和caspase-3的激活，呈时间和剂量依赖性。联合免疫沉淀实验显示，UMI-77阻断细胞中Mcl-1/Bax和Mcl-1/Bak的异源二聚化，从而拮抗Mcl-1的功能。通过使用Bax和Bak缺乏的小鼠胚胎成纤维细胞，进一步证实了Bax/Bak依赖性诱导细胞凋亡。在体内BxPC-3异种移植模型中，UMI-77有效抑制肿瘤生长。Western blot分析显示，促凋亡标志物增强，survivin显著降低。总的来说，这些有希望的发现证明了Mcl-1抑制剂对PC的治疗潜力，值得进一步的临床前研究。
Using a high throughput screening (HTS) approach, we have identified and validated several small molecule Mcl-1 inhibitors (SMIs). Here we describe a novel selective Mcl-1 SMI inhibitor, 2 (UMI-77), developed by structure-based chemical modifications of the lead compound 1 (UMI-59). We have characterized the binding of UMI-77 to Mcl-1 by using complementary biochemical, biophysical and computational methods, and determined its antitumor activity against panel of pancreatic cancer (PC) cells and in vivo xenograft model. UMI-77 binds to the BH3 binding groove of Mcl-1 with Ki of 490 nM, showing selectivity over other members of anti-apoptotic Bcl-2 members. UMI-77 inhibits cell growth and induces apoptosis in PC cells in a time and dose-dependent manner, accompanied by cytochrome c release and caspase-3 activation. Co-immunoprecipitation experiments revealed that UMI-77 blocks the heterodimerization of Mcl-1/Bax and Mcl-1/Bak in cells, thus antagonizing the Mcl-1 function. The Bax/Bak-dependent induction of apoptosis was further confirmed by using murine embryonic fibroblasts that are Bax and Bak deficient. In an in vivo BxPC-3 xenograft model, UMI-77 effectively inhibited tumor growth. Western blot analysis in tumor remnants revealed enhancement of pro-apoptotic markers and significant decrease of survivin. Collectively, these promising findings demonstrate the therapeutic potential of Mcl-1 inhibitors against PC and warrant further preclinical investigations.