Defining the relationship between kinase inhibitor pharmacophores and pro-/anti-targets using in vivo models of colorectal cancer

项目摘要 耐药性癌症是一个不幸的全球健康问题，影响着数百万人， 家庭[6]。靶向由受体激酶结构域突变引起的癌症的抑制剂 酪氨酸激酶通常靶向ATP结合位点[7]。虽然这种治疗方法是 通常非常有效的是，在“看门人”位置的突变和远离 药物结合位点不仅导致对药物的结合亲和力减弱， 在没有药物的情况下的激酶活性。解决额外的激酶激活是一个主要问题 因为无法治愈的癌症很容易继续扩散。 我的论文研究旨在了解和量化这些突变的影响 使用来自成纤维细胞的激酶结构域对激酶活化和热力学稳定性的影响 生长因子受体（FGFR）家族作为模型系统。最近，我们阐明了一种变构 FGFR激酶中从铰链区到激活环的残基网络， 与激活有关。使用NMR弛豫实验提供残留物特异性 信息，我计划量化在横纹肌肉瘤中发现的看门突变体的影响， 和铰链突变，最近发现严重削弱抑制剂结合，激酶动力学。 这些动力学将提供一个清晰的画面，这些突变如何劫持激酶的变构 网络来增加活性，并解释远离药物结合位点的铰链突变如何 赋予抵抗力。差示扫描量热法也将被用来了解药物如何 抗性突变影响激酶在载脂蛋白O和药物结合形式中的稳定性。的结果 这项工作将对未来的药物设计产生重大影响，以克服这些类型的药物 耐药突变

Project Summary Drug resistant cancers are an unfortunate global health problem that affects millions of families [6]. Inhibitors targeting cancers caused by mutations in the kinase domain of receptor tyrosine kinases often target the ATP binding site [7]. Although this method of treatment is generally very effective, mutations at the “gate-keeper” position and residues farther away from the drug binding site not only cause weakened binding affinity for the drug, but can also increase kinase activity in the absence of drug. Addressing additional kinase activation is a major concern as undruggable cancers can easily continue to spread. My dissertation research aims to understand and quantify the effects these mutations have on kinase activation and thermodynamic stability using the kinase domain from the fibroblast growth factor receptor (FGFRs) family as a model system. Recently, we elucidated an allosteric network of residues in FGFR kinases spanning from the hinge region to the activation loop that are associated with activation. Using NMR relaxation experiments to provide residue specific information, I plan to quantify the effects of the gate-keeper mutant found in rhabdomysarcoma, and a hinge mutant, recently found to severely weaken inhibitor binding, on kinase dynamics. These dynamics will provide a clear picture of how these mutations hijack the kinase's allosteric network to increase activity and explain how hinge mutations far from the drug-binding site can confer resistance. Differential scanning calorimetry will also be employed to understand how drug resistant mutations affect the stability of the kinase in the apo and drug-bound form. Findings from this work will have major implications for future drug design to overcome these types of drug resistance mutations.