Inhibition mechanism exploration of investigational drug TAK-441 as inhibitor against Vismodegib-resistant Smoothened mutant
Inhibition mechanism exploration of investigational drug TAK-441 as inhibitor against Vismodegib-resistant Smoothened mutant
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DOI：
10.1016/j.ejphar.2013.11.014
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发表时间：
2014-01-15
影响因子：
5
通讯作者：
Matsui, Hideki
中科院分区：
文献类型：
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作者：
Ishii, Tsuyoshi;Shimizu, Yuji;Matsui, Hideki
Hedgehog信号是髓母细胞瘤和基底细胞癌（BCC）的驱动力，使其成为有吸引力的治疗靶点。Vismodegib最近获得FDA批准用于治疗不可手术的BCC，但在临床研究中发现了耐药Smoothened（Smo）突变体（D473 H）。TAK-441是一种吡咯并[3，2-c]吡啶-4-酮衍生物，可有效抑制Rh信号转导，目前正在临床试验中进行研究。我们证明TAK-441抑制D473 H转染细胞中的报告基因活性，IC 50为79 nM，而Vismodegib显示IC 50 =7100 nM。为了研究抑制模式，我们采用三种不同的结合试验评价了Smo抑制剂，如[H-3]-TAK-441膜结合试验、亲和选择MS检测试验和二吡环巴胺全细胞试验。在三种不同的测定中，与野生型Smo相比，维莫德吉和环巴胺显示出对D473 H突变体的较低亲和力。另一方面，在结合试验中，尽管TALK 441与其他两种众所周知的抑制剂结合相同的结合位点，但与野生型Smo相比，TAK-441对D473 H突变体的结合亲和力几乎相等。这些体外研究结果表明。TALK 441具有在依赖于Hedgehog信号传导的癌症中临床使用的潜力，包括野生型肿瘤和维莫德吉抗性。D473 H突变体。(C)2013爱思唯尔有限公司版权所有。
Hedgehog signaling is a driving force in medulloblastoma and basal cell carcinoma (BCC), making if an attractive therapeutic target. Vismodegib recently received FDA approval for the treatment of inoperable BCC, but a drug-resistant Smoothened (Smo) mutant (D473H) was identified in a clinical study. TAK-441 is a pyrrolo[3,2-c]pyridine-4-one derivative that potently inhibits Rh signal transduction and is currently under investigation in clinical trials. We demonstrated that TAK-441 inhibits reporter activity in D473H-transfected cells with an IC50 of 79 nM, while Vismodegib showed an IC50=7100 nM. In order to investigate the mode of inhibition, we evaluated the Smo inhibitors with three different binding assays, such as]H-3]-TAK-441 membrane binding assay, affinity selection MS detection assay, and bodipycylopamine whole cell assay. In three different assays, Vismodegib and cyclopamine showed lower affinity for the D473H mutant in comparison with wild type Smo. On the other hand, TAK-441 showed almost equal binding affinity for the D473H mutant compared with wild type Smo in the binding assays, although TALK 441 binds to the same binding site as two other well-known inhibitors. These in vitro findings suggest. that TALK 441 has the potential for clinical use in cancers that are dependent on Hedgehog signaling, including wild type tumors and Vismodegib-resistant. D473H mutants. (C) 2013 Elsevier B.V. All rights reserved.