A transformative drug discovery platform for allosteric kinase inhibitors

项目摘要 明尼苏达大学(UMN)和位于明尼苏达州的药物发现初创公司光子制药(PP)已经 合作优化、现场测试和工业规模部署，这是一种创新的开发新方法 变构酶抑制剂(AKI)。这些分子具有很高的潜力作为新的癌症治疗药物 避免临床对常规正构体酶抑制剂(OKI)的耐药性。我们已经发展出了高度- 基于Förster纳秒荧光寿命检测的吞吐量筛选(HTS)技术 共振能量转移(FRET)，跟踪配体驱动的激酶变构，通过 监测激活环的结构变化，激活环是所有激酶中的关键调节元件。这是第一次 HTS顺从技术，依赖于高质量的，准确地解决了激酶抑制剂的变构效应 传统的荧光平板阅读器(PR)无法读取纳秒的Flt读数。PP已经发展起来 专利HTS平台，使用FRET生物传感器和最先进的Flt-PR来检测结构 对于384孔板，读数为2分钟；对于1536孔板，读数为5分钟。通过与PP合作，我们将转变我们的 将激酶FRET传感器技术转化为一个广泛适用的药物发现平台，用于识别Akis。 我们针对两个不同的目标提出了药物发现计划，以展示广泛的实用性并加速大规模的 大规模采用我们的药物开发技术。在AIM 1中，我们发现了Aurora A抑制剂 通过抑制Aurora A与c-Myc的支架相互作用下调不可抑制的c-Myc癌蛋白 MYC。这些分子将代表一种新的治疗策略，用于治疗大量患有 C-Myc驱动的肿瘤。在AIM 2中，我们发现c-met受体酪氨酸激酶的变构抑制剂是一种新的 C-met基因驱动的肺癌患者的治疗策略。这些患者总是对 目前的MET抑制剂通过获得的ATP位点突变，以及结合在细胞外的变构抑制剂 ATP-SITE将绕过这种耐药模式，填补一个未得到满足的临床需求。这一UMN-PP伙伴关系 将两位世界领先的专家-莱文森和托马斯-数十年的生物物理学研究转化为药物- 通过分解激酶中的配体驱动的变构来发现。这是由FLT-PR工具和 在真正分解变构抑制剂的分析中实施纳秒Flt检测所需的技术诀窍 HTS模式。这克服了传统的激酶抑制物筛查的关键缺陷，即检测激酶 抑制或结合而不考虑变构机制。该平台具有广泛的适用性，因为几乎所有 我们的技术检测到了激酶经历了大规模的变构结构变化。这件事成功了 该项目将促进采用这项技术，目标是广泛的生物医学重要的激酶，如 Photonic Pharma与百时美施贵宝在其他药物研发方面的成功合作 高度优先的治疗目标。

Project Summary The University of Minnesota (UMN) and Photonic Pharma (PP) a Minnesota-based drug discovery start-up, have partnered to optimize, field-test, and deploy at industrial scale, an innovative new approach to developing allosteric kinase inhibitors (AKI). These molecules have high potential as novel cancer therapeutics that circumvent clinical resistance to conventional orthosteric kinase inhibitors (OKI). We have developed high- throughput screening (HTS) technology based on nanosecond fluorescence lifetime (FLT) detection of Förster resonance energy transfer (FRET), that tracks ligand-driven kinase allostery with angstrom precision by monitoring structural changes of the activation loop, the key regulatory element in all kinases. This is the first HTS-amenable technology that accurately resolves allosteric effects of kinase inhibitors, relying on high-quality nanosecond FLT readouts unavailable from conventional fluorescence plate readers (PR). PP have developed a proprietary HTS platform that uses FRET biosensors and a state-of-the-art FLT-PR to detect structural readouts in <2 min for 384-well and <5 min for 1536-well plates. By partnering with PP, we will transform our kinase FRET sensor technology into a broadly applicable drug-discovery platform for identifying AKIs. We propose drug-discovery programs on two different targets to demonstrate broad utility and accelerate large- scale adoption of our technology for drug development. In AIM 1, we identify Aurora A inhibitors that downregulate the undruggable c-Myc oncoprotein by inhibiting the scaffolding interaction of Aurora A with c- Myc. These molecules would represent a novel treatment strategy for the large number of cancer patients with c-Myc-driven tumors. In AIM 2, we identify allosteric inhibitors of the c-MET receptor tyrosine kinase as a novel therapeutic strategy for patients with c-MET-driven lung cancer. These patients invariably develop resistance to current MET inhibitors through acquired mutations in the ATP site, and allosteric inhibitors that bind outside the ATP-site would circumvent this mode of resistance, filling an unmet clinical need. This UMN-PP partnership translates decades of biophysics research by two world-leading experts – Levinson and Thomas – toward drug- discovery by resolving ligand-driven allostery in kinases. This is enabled by the FLT-PR instrumentation and know-how required to implement nanosecond FLT detection in assays that resolve allosteric inhibitors in true HTS mode. This overcomes key drawbacks of conventional kinase inhibitor screens, which detect kinase inhibition or binding without regard to allosteric mechanism. The platform is broadly applicable, as almost all kinases undergo the large-scale allosteric structural changes our technology detects. Success of this project will catalyze adoption of this technology targeting a wide range of biomedically important kinases, as highlighted by Photonic Pharma’s successful partnership with Bristol-Myers Squibb on drug discovery for other high-priority therapeutic targets.