Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source
Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source
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DOI：
10.3791/62414
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发表时间：
2021-05-01
影响因子：
1.2
通讯作者：
von Delft, Frank
中科院分区：
文献类型：
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作者：
Douangamath, Alice;Powell, Ailsa;von Delft, Frank
在基于片段的药物发现中，针对感兴趣的蛋白质测试数百种或通常数千种小于300 Da的化合物，以鉴定可以开发成有效候选药物的化学实体。由于化合物很小，相互作用很弱，因此筛选方法必须高度敏感;此外，结构信息往往是将这些命中物加工成铅样化合物的关键。因此，蛋白质晶体学一直是一项黄金标准技术，但历史上太具有挑战性，无法作为初级筛选广泛使用。最初的XChem实验在2014年进行了演示，然后与学术和工业合作者进行了试验，以验证该过程。从那时起，大量的研究工作和大量的射束时间简化了样品制备，开发了具有快速后续可能性的片段库，自动化并提高了I 04 -1射束线的无人值守数据收集能力，并实施了用于数据管理，分析和命中识别的新工具。XChem现在是一个大规模晶体碎片筛选的设施，支持整个晶体沉积过程，并可供世界各地的学术和工业用户使用。自2016年以来，经过同行评审的学术用户计划一直在积极开发，以适应尽可能广泛的科学范围的项目，包括经过验证的以及探索性的项目。学术访问是通过一年两次的呼吁同行评审的建议，和专有工作是由钻石的工业联络组安排。该工作流程已被常规应用于来自不同治疗领域的100多个靶标，并有效识别弱结合剂（1%-30%命中率），这既是化合物设计的高质量起点，也提供了关于结合位点的广泛结构信息。在COVID-19大流行期间持续筛选SARS-CoV-2靶标，包括主要蛋白酶的3周周转，证明了该过程的弹性。
In fragment-based drug discovery, hundreds or often thousands of compounds smaller than similar to 300 Da are tested against the protein of interest to identify chemical entities that can be developed into potent drug candidates. Since the compounds are small, interactions are weak, and the screening method must therefore be highly sensitive; moreover, structural information tends to be crucial for elaborating these hits into lead-like compounds. Therefore, protein crystallography has always been a gold-standard technique, yet historically too challenging to find widespread use as a primary screen.Initial XChem experiments were demonstrated in 2014 and then trialed with academic and industrial collaborators to validate the process. Since then, a large research effort and significant beamtime have streamlined sample preparation, developed a fragment library with rapid follow-up possibilities, automated and improved the capability of I04-1 beamline for unattended data collection, and implemented new tools for data management, analysis and hit identification.XChem is now a facility for large-scale crystallographic fragment screening, supporting the entire crystals-to-deposition process, and accessible to academic and industrial users worldwide. The peer-reviewed academic user program has been actively developed since 2016, to accommodate projects from as broad a scientific scope as possible, including well-validated as well as exploratory projects. Academic access is allocated through biannual calls for peer-reviewed proposals, and proprietary work is arranged by Diamond's Industrial Liaison group. This workflow has already been routinely applied to over a hundred targets from diverse therapeutic areas, and effectively identifies weak binders (1%-30% hit rate), which both serve as high-quality starting points for compound design and provide extensive structural information on binding sites. The resilience of the process was demonstrated by continued screening of SARS-CoV-2 targets during the COVID-19 pandemic, including a 3-week turnaround for the main protease.