Target enablement

该中心药物发现计划的成功有赖于能够在实验中支持 复合型发展。在Target Enablement项目中，我们将开发和验证实验 程序通过制作“目标使能包”(TEP)。这些协议包括一套实验方案 以及产生快速、可靠、全频谱读数的输出，这些读数支持设计制造的加速- 测试-分析(DMTA)周期。TEP是靶标和靶点上药物化学的高效促进剂 以可以实现的方式为目标；然而，它们往往仍然不完整，因为它们 需要很大的努力来从文献和/或反复试验中收集资料；并确定药物 不管怎样，化学家们可能会设法使化合物系列取得进展。然而，这掩盖了现实和机会 这样做的成本，包括合成不必要的化合物，追求死胡同的假设， 以及糟糕的数据模型。在Target Enablement项目中，我们建议生成下一代TEP 其中包括用于产生生物化学行为良好的结晶蛋白质的试剂和方案，3D 晶体碎片筛选结合位点(S)的相互作用图、生化和生物物理分析 和有说服力的主打化合物，表现出一致的低微摩尔亲和力、活性和结合姿势， 在一组类比上是合理的。我们将通过以下方式提供严格、有效和可重现的TEP (A)采用牛津大学自2015年以来制定的既定、系统的工作流程；和(B) 实施和强化最近的创新和新技术，包括在 COVID登月和其他与大流行有关的工作的尾声。因此，项目2的具体目标是生成 针对10个目标的新一代TEP，以推动项目3中快速实现从点击到领先的进展，并由此验证 病毒学和化学生物学提出的抗病毒假说。目标分为4个目标，其中目标1确立 项目1中确定的所有新目标的生物化学和晶体结构可控性。目标2建立在产出的基础上 来自目标1，并开发了健壮的结晶方案以及生化活性和 生物物理亲和力。目标3需要完成和分析晶体碎片筛选。目标4 通过验证分析并将姿势与从观察到的碎片中提取的命中化合物结合来最终确定TEP。在……里面 摘要，项目2将已建立的工作流程与最近的创新相结合，以提供严格的TEP， 有效和可重复的，为项目3的深远创新提供基础，从而确保 尽快在预算范围内按时交付。此外，还将迅速发布临时工作方案，以便开展并行工作。 并增加为大流行做好准备的总体机会。该项目将是 由钻石光源公司的同一个团队完成了破纪录的结晶学研究 (XChem)播种COVID月球快照的碎片屏幕。

The success of the Center's drug discovery programs relies on being experimentally fully enabled to support compound development. In the Target Enablement project, we will develop and validate the experimental procedures by producing "Target Enabling Packages" (TEPs). These comprise a set of experimental protocols and outputs that generate the rapid, reliable, full-spectrum readouts that underpin the acceleration of the designmake- test-analyze (DMTA) cycles. TEPs are highly effective enablers of medicinal chemistry on targets and targeting modalities where they can be achieved; nevertheless, frequently they remain incomplete, since they require great effort to assemble from literature and/or trial-and-error experiments; and determined medicinal chemists might manage to progress compound series anyway. However, this conceals the real and opportunity costs of doing so, which includes the synthesis of unnecessary compounds, pursuit of dead-end hypotheses, and poor models of the data. In the Target Enablement project, we propose to generate next generation TEPs that include the reagents and protocols for generating biochemically-behaved, well-crystallizing protein, 3D interaction maps of the binding site(s) from a crystal-based fragment screen, biochemical and biophysical assays and persuasive hit compounds, displaying consistent low micromolar affinity, activity and binding pose, rationalizable across a set of analogues. We will deliver TEPs that are rigorous, effective, and reproducible, by (a) contracting in the established, systematic workflow developed since 2015 at the University of Oxford, and (b) implementing and hardening of recent innovations and new technologies, including methods developed in the wake of the COVID Moonshot and other pandemic-related work. The specific goal of Project 2 is thus to generate next generation TEPs for 10 targets, to drive rapid hit-to-lead progression in Project 3 and thence validation of the antiviral hypothesis by virology and chemical biology. The goal is spread over 4 Aims, with Aim 1 establishing biochemical and crystal structure tractability of all novel targets identified in Project 1. Aim 2 builds on the outputs from Aim 1 and develops the robust crystallization protocols, and orthogonal assays for biochemical activity and biophysical affinity. Aim 3 entails the completion and analyses of the crystallographic fragment screens. Aim 4 finalizes TEPs by validating assays and binding pose with hit compounds derived from observed fragments. In summary, Project 2 combines an established workflow with recent innovations to deliver TEPs that are rigorous, effective and reproducible, providing the foundations for the far-reaching innovations of Project 3 thus ensuring ASAP can deliver on time and on budget. Moreover, TEPs will be promptly published to enable parallel efforts beyond ASAP and increase the overall chances of achieving pandemic preparedness. The project will be delivered by the same team at Diamond Light Source that executed the record-breaking crystallographic (XChem) fragment screen that seeded the COVID Moonshot.