Targeting the GTP-bound state of oncogenic Ras proteins

研究总结/摘要 Ras基因突变是人类癌症最常见的激活剂，也是癌症的有效抑制剂。 它们编码的GTP酶已被寻求作为潜在的癌症治疗剂。尽管在这方面进行了广泛的研究， Ras蛋白仍然是医学化学中最具挑战性的靶标之一。中的致癌突变 Ras基因通常导致蛋白质锁定在“开启”GTP结合状态，从而结合其效应物Raf激酶 PI3K因此，靶向GTP结合Ras蛋白并破坏与效应物相互作用的小分子 有望成为有前景的癌症治疗剂。该提案寻求开发小分子抑制剂， Ras（GTP）通过靶向最近发现的“开关II沟”。该战略结合了以下技术 合成化学、组合化学和化学生物学，以增加 结合剂，评估它们作为Ras（GTP）抑制剂的作用，并开发N-和H-Ras癌基因选择性抑制剂。 这项研究策略的目的是利用我现有的合成化学知识和培训 新的化学生物学技术。实验工作需要我学习蛋白质的表达， 纯化、蛋白质NMR光谱和一系列生化测定来检测Ras GTP酶的抑制。 通过阅读相关文献，并与有经验的科学家讨论这个项目，我将大大提高 我对参与细胞生长和增殖的蛋白质信号通路的理解是错误的， 调节癌症。加州大学旧金山分校是一个理想的位置，我接受培训的化学生物学，由于其广泛的范围 经验丰富的教师，优秀的仪器设备，并与更广泛的化学生物学连接 社区 我打算从事应用有机化学来改善人类健康的学术事业; 具体来说，我希望开发新的合成方法，以发现具有挑战性的抑制剂。 和动态蛋白质靶点。我的学术研究和研究生研究给了我很强的背景 在有机化学和合成方法方面，我缺乏化学生物学方面的正规训练。我选择 我加入了加州大学旧金山分校Shokat教授的团队，因为我相信他是一个理想的导师，可以帮助我弥补这一不足 更好地为我的独立事业做准备

RESEARCH SUMMARY/ABSTRACT Mutations in Ras genes are the most common activators of cancer in humans, and effective inhibitors of the GTPases they encode have been sought as potential cancer therapeutics. Despite extensive research in this field, Ras proteins remain among the most challenging targets in medical chemistry. Oncogenic mutations in Ras genes typically result in proteins locked in the “on” GTP-bound state, which binds its effectors Raf kinases and PI3K. Therefore, small molecules that target GTP-bound Ras proteins and disrupt interactions with effectors are expected to be promising cancer therapeutics. This proposal seeks to develop small molecule inhibitors of Ras(GTP) by targeting the recently discovered “switch II groove.” The strategy combines techniques from synthetic chemistry, combinatorial chemistry, and chemical biology to increase the non-covalent affinity of binders, evaluate them as inhibitors of Ras(GTP), and develop N- and H-Ras oncogene-selective inhibitors. This research strategy was designed to both leverage my existing knowledge of synthetic chemistry and train me in new chemical biology techniques. The experimental work will require me to learn protein expression and purification, protein NMR spectroscopy, and a range of biochemical assays to detect inhibition of Ras GTPases. By reading the relevant literature and discussing this project with experienced scientists, I will greatly improve my understanding of the protein signaling pathways involved in cell growth and proliferation which are mis- regulated in cancer. UCSF is an ideal location for me to receive training in chemical biology due to its wide range of experienced faculty, excellent instrumentation facilities, and connections with the wider chemical biology community. I intend to pursue an academic career applying organic chemistry to the improvement of human health; specifically, I wish to develop new synthetic methodology to enable the discovery of inhibitors for challenging and dynamic protein targets. My academic studies and graduate research have given me a strong background in organic chemistry and synthetic methods; however, my formal training in chemical biology is lacking. I chose to join Professor Shokat’s group at UCSF because I believe he is an ideal mentor to help me remedy that lack and better prepare me for an independent career.