A high-throughput platform for crystallography-based fragment screening

项目摘要/摘要 基于片段的药物发现(FBDD)是制药行业广泛使用的一种药物发现方法 Novo设计的针对新药候选药物的分子。FBDD允许更有效地探索 与高通量筛选相比，化学空间具有更高的命中率，这可以有显著的意义 在早期药物发现和在具有挑战性或“不可下药”的目标的情况下的影响。FBDD导致了 大约30种新药进入临床试验，2种已经进入市场。FBDD还可用于 为已经上市的经过充分验证的重要药物靶点发现和开发新分子 针对它们的药物，既提高了疗效又降低了毒性，并创造了新的智力 非专利药品的财产。蛋白质x射线结晶学(Px)是确定 与药物靶标结合的给定片段的3D位置和方向。PX还可以检测到更大范围的 与用于片段和化合物筛选的其他生物物理方法和IS的结合亲和力比较 与蛋白质大小无关。然而，结晶学对于筛选大的 由于大规模生产共晶晶体的显著瓶颈，片段文库 碎片浸泡、晶体采集、X射线数据采集、结构测定和分析。 互补的生物物理技术经常被用来预先筛选结合的片段，然后PX 在第二步中用于确定每个片段的准确结合姿势。Accelero BioStructures是 开发一个首次上市的、高效的、基于PX的一步法片段文库筛选平台，该平台可以 通过大幅提高开发新引线的效率和降低成本来彻底改变该领域 用于临床前试验的分子。在第一阶段，我们评估了一种高密度结晶网格， 提高了靶-碎片共结晶、碎片浸晶和同步加速器的效率- 基于数据收集，单步命中率约为5%，同时生成3D细节 蛋白质-片段相互作用。在成功完成第一阶段目标后，我们现在正在推进 我们的第二阶段计划将这项实验技术与分布式计算结晶学相结合 管道和数据管理/信息学主干，这将使我们能够高效地处理大片段 存储库屏幕。我们将使用几个可用药和不可用药的肿瘤学靶点，这些靶点与各种 来自我们的行业和学术客户的癌症，作为概念验证系统来展示 我们的整体平台。我们的计划与NCATS的所有药物发现和开发SBIR保持良好的一致性 感兴趣的主题：“工具和技术，使分析化合物的活动目前”非 可药物的“靶点”；“共结晶高通量筛选技术”；“工具和技术， 提高药物化学、生物或其他干预措施优化的预测性或效率“；以及 开发高通量成像技术，使翻译研究变得更多 有效率“。

Project Summary/Abstract Fragment-based drug discovery (FBDD) is a widely used method in the pharmaceutical industry for the de novo design of molecules that target new drug candidates. FBDD allows a more effective exploration of chemical space with a higher hit rate compared to high-throughput screening, and this can have significant effects in early drug discovery and in the case of challenging or “non-druggable” targets. FBDD has led to around 30 new drugs entering clinical trials and 2 that have entered the market. FBDD can also be used to discover and develop novel molecules for well-validated and important drug targets that already have marketed drugs against them, both for increasing efficacy with lower toxicity as well as creation of new intellectual property for off-patent drugs. Protein x-ray crystallography (PX) is the gold standard for determining the exact 3D location and orientation of a given fragment bound to a drug target. PX can also detect a wider range of binding affinities compared to other biophysical methods for fragment and compound screening and is independent of protein size. However, crystallography is expensive and inefficient for screening a large fragment library due to significant bottlenecks in mass production of crystals for co-crystallization, crystal soaking with fragments, crystal harvesting, X-ray data collection, structure determination and analysis. Complementary biophysical techniques are often used to prescreen for fragments that bind and PX is then used in a second step to determine the exact binding pose of each fragment. Accelero Biostructures is developing a first-to-market, efficient, one-step PX-based fragment library-screening platform that can revolutionize the field by dramatically increasing the efficiency and reducing the cost of developing novel lead molecules for preclinical testing. In Phase I we evaluated a high-density crystallization grid that dramatically increased the efficiency of target-fragment co-crystallization, crystal soaking with fragments and synchrotron- based data collection, leading to a hit rate of ~5% in a single step while simultaneously producing 3D details of protein-fragment interactions. After successfully completing our Phase I aims, we are now moving ahead with our Phase II plan to integrate this experimental technology with a distributed computational crystallography pipeline and data management/informatics backbone that will allow us to efficiently process a large fragment library screen. We will use several druggable and non-druggable oncology targets implicated in various cancers, from our industry and academic customers, as proof-of-concept systems to demonstrate the utility of our overall platform. Our plans are well-aligned with all of NCATS Drug Discovery and Development SBIR topics of interest: “Tools and technologies to enable assaying of compound activity on currently “non- druggable” targets”; “Co-crystallization high-throughput screening techniques”; “Tools and technologies that increase the predictivity or efficiency of medicinal chemistry, biologic or other intervention optimization”; and “Development of high-throughput imaging technologies that focus on making translational research more efficient”.