New Affinity Probe Targeting VEGF Receptors for Kinase Inhibitor Selectivity Profiling by Chemical Proteomics
New Affinity Probe Targeting VEGF Receptors for Kinase Inhibitor Selectivity Profiling by Chemical Proteomics
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DOI：
10.1021/pr401247t
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发表时间：
2014-05-01
影响因子：
4.4
通讯作者：
Kuster, Bernhard
中科院分区：
文献类型：
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作者：
Ku, Xin;Heinzmeir, Stephanie;Kuster, Bernhard
实体瘤的生长依赖于营养和氧气供应，它们通常通过新生血管生成获得。血管内皮生长因子和相应的受体（VEGFRs）在这一过程中起着核心作用，因此，阻断这一途径是癌症治疗的一种治疗策略。许多抑制VEGFR的小分子抑制剂已被开发用于临床应用，全面了解靶点选择性对于评估药物分子的治疗效果和潜在风险非常重要。基于质谱的化学蛋白质组学的最新进展允许在接近生理条件下分析药物靶标相互作用，在本研究中，我们报告了小分子亲和探针的设计、合成和应用，作为VEGFR和其他激酶抑制剂选择性分析的工具。该探针能够结合bbb132蛋白激酶，包括来自癌细胞系或人胎盘组织裂解物的血管激酶，如VEGFRs、PDGFRs和c-KIT。将新探针与Kinobeads结合进行竞争结合试验，我们能够鉴定出VEGFR/PDGFR抑制剂pazopanib和axitinib的纳米级脱靶。由于其广泛的结合谱，所开发的化学工具可用于发现激酶抑制剂靶点，这可能有助于更全面地了解此类药物的作用机制。
Solid tumors are dependent for growth on nutrients and the supply of oxygen, which they often acquire via neoangiogenesis. Vascular endothelial growth factors and the corresponding receptors (VEGFRs) play central roles in this process, and consequently, the blockade of this pathway is one therapeutic strategy for cancer treatment. A number of small molecules inhibiting VEGFR inhibitors have been developed for clinical use, and a comprehensive view of target selectivity is important to assess the therapeutic as well as risk potential of a drug molecule. Recent advances in mass spectrometry-based chemical proteomics allow analyses of drug target interactions under close-to-physiological conditions, and in this study, we report on the design, synthesis, and application of a small molecule affinity probe as a tool for the selectivity profiling of VEGFR and other kinase inhibitors. The probe is capable of binding >132 protein kinases, including angiokinases such as VEGFRs, PDGFRs, and c-KIT from lysates of cancer cell lines or human placenta tissue. Combining the new probe with Kinobeads in competitive binding assays, we were able to identify nanomolar off-targets of the VEGFR/PDGFR inhibitors pazopanib and axitinib. Because of its broad binding spectrum, the developed chemical tool can be generically used for the discovery of kinase inhibitor targets, which may contribute to a more comprehensive understanding of the mechanisms of action of such drugs.