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

项目摘要 自从伊马替尼（一种用于治疗慢性髓性白血病的特异性激酶抑制剂）问世以来， 小分子激酶抑制剂的开发是对抗癌症的有希望的途径。特别是， 细胞周期蛋白依赖性激酶2（Cdk 2）是一个很有前途的抗癌靶点，因为它通常被细胞周期蛋白激活， E在G1/S期转变时进行细胞周期，并在疾病中过度激活，例如 卵巢癌乳腺癌和结肠直肠癌从历史上看，靶向Cdk 2涉及设计结合其受体的分子。 活性位点，由于激酶之间ATP结合位点的同源性，导致许多脱靶效应， 尤其是其它Cdk酶。然而，我们的团队最近发现并表征了一种变构结合， Cdk 2中的口袋与染料8-苯胺基-1-萘磺酸（ANS）结合。没有证据表明这 口袋存在于其他Cdk家庭成员，因此提供了一个杰出的机会，发现真正的 选择性一流的Cdk 2抑制剂，可能没有脱靶毒性。通过靶向变构 口袋，我们假设，新的抑制剂可以开发，将不结合其他Cdk家族成员 考虑到该变构位点的非同源性。目前还没有已知的有效的变构剂， Cdk 2的选择性抑制剂。 我的建议集中在开发新的变构抑制剂使用两种方法：化学库 ANS类似物的开发（目标1）和结合变构口袋的新化学物质的发现 使用高通量筛选（HTS），片段筛选，虚拟筛选和确认，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，以提供化学和结构上的 对开发一种有效的和选择性的Cdk 2变构抑制剂来治疗癌症的见解。如果 成功，这一系列的策略可以作为一个例子，激酶抑制剂的设计一般。这 奖学金提案还将提供药物化学、化学生物学和翻译方面的培训。 科学，作为一名欣赏药物各个方面的医生科学家，为有意义的职业生涯做出贡献 发展过程这项培训将使我成为一名独立的研究人员， 发现新疗法的科学家和测试它们的医生之间的差距。这项工作 这与NCI的使命一致，即将新的治疗方法纳入临床实践。

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.