Development of Allosteric Inhibitors against Cyclin-Dependent Kinase 2 (Cdk2)

项目摘要 自伊马替尼问世以来，伊马替尼是一种用于治疗慢性粒细胞白血病的特异性激酶抑制剂，进一步 开发小分子激酶抑制剂一直是抗击癌症的一条很有前途的途径。特别是， 细胞周期蛋白依赖性激酶2(Cdk2)是一种很有前途的抗癌靶点，因为它通常被细胞周期蛋白激活。 通过细胞周期的G1/S相转变，并在疾病，如过度激活 卵巢癌、乳腺癌和结直肠癌。从历史上看，以CDK2为靶点包括设计与其结合的分子 活性部位，导致许多脱靶效应，由于同源性的三磷酸腺苷结合位点之间的激酶， 尤其是其他CDK酶。然而，我们的团队最近发现并表征了一种变构结合 口袋中的Cdk2与染料8-苯胺基-1-萘磺酸(ANS)结合。没有证据表明这一点 Pocket存在于CDK家族的其他成员中，因此提供了一个绝佳的机会来真正发现 选择性的一流的CDK2抑制剂，可能没有靶外毒性。通过靶向变构 我们假设可以开发出不与其他CDK家族成员结合的新型抑制剂 考虑到这个变构位点的非同源性。目前还没有已知的有效的变构也没有 CDK2选择性抑制剂。 我的建议集中在利用两种方法开发新型变构抑制剂：化学库 ANS类似物的开发(目标1)和发现结合变构口袋的新化学物质 使用高通量筛选(HTS)、片段筛选、虚拟筛选和通过 生物测试(目标2)。这些技术提供了一种多管齐下的方法来提供化学和结构 对开发有效和选择性的CDK2变构抑制剂治疗癌症的洞察。如果 如果成功，这一系列的策略可以作为一般的激酶抑制剂设计的范例。这 奖学金计划还将提供药物化学、化学生物学和翻译方面的培训。 科学，有助于作为一名内科科学家的有意义的职业生涯，他欣赏药物的方方面面 发展进程。这次培训将使我成为一名独立的研究员，能够在 发现新疗法的科学家和测试它们的医生之间的差距。这部作品 与NCI将新疗法纳入临床实践的使命非常一致。

Project Summary Since the advent of imatinib, a specific kinase inhibitor used to treat chronic myeloid leukemia, further development of small molecule kinase inhibitors has been a promising avenue to combat cancer. In particular, cyclin-dependent kinase 2 (Cdk2) has been a promising anti-cancer target, as it is normally activated by cyclin E to proceed through the cell cycle at the G1/S phase transition, and hyperactivated in diseases such as ovarian, breast, and colorectal cancers. Historically, targeting Cdk2 involved designing molecules that bind its active site, leading to many off-target effects due to the homology of the ATP-binding site among kinases, especially other Cdk enzymes. However, our group recently discovered and characterized an allosteric binding pocket in Cdk2 bound to the dye 8-anilino-1-naphthalenesulfonic acid (ANS). There is no evidence that this pocket exists in other Cdk family members and therefore provides an outstanding opportunity to discover truly selective first-in-class Cdk2 inhibitors that could be devoid of off-target toxicity. By targeting the allosteric pocket, we hypothesize that novel inhibitors can be developed that will not bind to other Cdk family members given the non-homologous nature of this allosteric site. There is currently no known potent allosteric nor selective inhibitor of Cdk2. My proposal focuses on developing novel allosteric inhibitors using two approaches: chemical library development of ANS analogs (Aim 1) and discovery of new chemical matter that binds the allosteric pocket using high-throughput screening (HTS), fragment screening, virtual screening, and confirmation through biological testing (Aim 2). These techniques offer a multipronged approach to provide chemical and structural insight towards the development of a potent and selective allosteric inhibitor of Cdk2 to treat cancer. If successful, this array of strategies can serve as an example for kinase inhibitor design in general. This fellowship proposal will also provide training in medicinal chemistry, chemical biology, and translational science, contributing to a meaningful career as a physician scientist who appreciates all facets of the drug development process. This training will allow me to become an independent researcher who can bridge the gap between the scientists who discover new therapeutics and the physicians who test them. This work strongly aligns with the mission of NCI to incorporate novel treatments into clinical practice.