Multiplexed Nucleation Approaches for Enhanced High Throughput Screening of Co-Crystals

项目摘要 DeNovX的技术改善了活性药物成分（API）和蛋白质的结晶。超过 70%的原料药表现出水溶性和生物利用度差，导致药物失效。共结晶 将API与超分子杂合子配对，并且是产生H2O的晶体工程方法 可溶性API组合物，但其对于高通量筛选（HTS）尚不可再现。一种冲压机/模具， HTS格式和液压机可用于压缩机械化，以提供可重复的 足以形成共晶的剪切力。第一阶段证明了具有48孔格式的高置信度POC 用于API共晶体的HTS机械结晶，并在连续变化中给出了优异的再现性 study.溶剂滴研磨基准与压缩机械研磨的比较表明， 后者可以在减少81%的时间内进行，减少60%的材料，同时产生25%的样品， 分析.通过分奖项给斯坦福大学的同步辐射光源（SSRL）和阿贡的 先进光子源（APS），第二阶段将集成高温超导机械化与同步加速器粉末 X射线衍射（PXRD）分析，提供无与伦比的次要成分鉴定、定量、结构， 和吞吐量。具体目标1：使用以下器械进行压缩机械化的重复（n ≥ 6）研究 α-原型用于识别对API共结晶影响最大的变量。检查两个基准和≥ 14 来自酸性、碱性和中性API类的共晶体与具有以下特征的杂环化合物适当匹配： 互补氢键行为。通过SSRL和APS的子授权收集同步加速器PXRD。具体目标 2：测试≥ 6个压缩机械化试验台-原型，可用作消耗品样品架，用于 API和共晶体的PXRD分析。四个原型与同步加速器PXRD兼容， 适用于实验室PXRD的低复用格式。API共晶样品在多重支架中， n ≥ 6次连续变化研究的同步加速器PXRD组成在平均值±3σ内。具体目标3： 进行高通量同步加速器PXRD数据采集，以证明以下物质的检测限≤ 0.2%（w/w） 在连续变化研究中，使用预成型-原型机械结晶样品的次要组分API相 持有人同时证明每个样品的PXRD图谱采集速率≤ 90 s，同时保留数据 质量.具体目标4：证明以下物质的纯的和溶剂稀疏的压缩机械化HTS： 在SSRL/APS采用同步加速器PXRD对≥ 6个高冲击API靶点中的每一个进行共晶， 制药公司和NIH。确定新的共晶相和制备条件， 利益相关者的溶解度/渗透性研究。用于原料药HTS机械化的可复制工具， 共晶体将通过创造新的或再利用的API组合物而有益于公共健康， 体内溶解度和生物利用度，为1万亿美元的药品市场。

PROJECT SUMMARY DeNovX’s technologies improve crystallization of active pharmaceutical ingredients (APIs) and proteins. Over 70% of APIs exhibit poor H2O solubility and bioavailability that contribute to drug failures. Co-crystallization mates an API with a supramolecular heterosynthon and is a crystal engineering approach to creating H2O soluble API compositions, but it is not yet reproducible for high throughput screening (HTS). A punch/die in an HTS format and a hydraulic press can be used for compressive mechanocrystallization to give reproducible shear forces adequate to form co-crystals. Phase I demonstrated a high confidence POC with a 48 well format for HTS mechanocrystallization of an API co-crystal and gave excellent reproducibility in a continuous variation study. A comparison of the solvent drop grinding benchmark with compressive mechanocrystallization showed that the latter can be conducted in 81% less time with 60% less material while yielding 25% more sample for analysis. Through subawards to Stanford’s Synchrotron Radiation Lightsource (SSRL) and Argonne’s Advanced Photon Source (APS), Phase II will integrate HTS mechanocrystallization with synchrotron powder X-ray diffraction (PXRD) analysis to give unparalleled minor constituent identification, quantitation, structure, and throughput. Specific Aim 1: Conduct replicate (n ≥ 6) studies of compressive mechanocrystallization using α-prototypes to identify variables most impacting API co-crystallization. Examine two benchmarks and ≥ 14 co-crystals from the acidic, basic, and neutral API classes matched appropriately to heterosynthons having complementary H-bonding behavior. Collect synchrotron PXRD by subawards to SSRL and APS. Specific Aim 2: Test ≥ 6 compressive mechanocrystallization ꞵ-prototypes that can serve as consumable sample holders for PXRD analyses of APIs and co-crystals. Four prototypes to be compatible with synchrotron PXRD and two lower multiplexed formats suitable for laboratory PXRD. API co-crystal samples in multiplexed holders to give synchrotron PXRD compositions within ±3σ of averages for n ≥ 6 continuous variation studies. Specific Aim 3: Conduct high throughput synchrotron PXRD data collection to demonstrate limit of detection ≤ 0.2% (w/w) for minor constituent API phases in a continuous variation study using 𝛾-prototype mechanocrystallization sample holders. Concurrently demonstrate PXRD pattern acquisition rates ≤ 90 s per sample while retaining data quality. Specific Aim 4: Demonstrate neat and solvent sparse compressive mechanocrystallization HTS of co-crystals with synchrotron PXRD at SSRL/APS for each of ≥ 6 high impact API targets relevant to pharmaceutical companies and NIH. Identify new co-crystal phases and preparative conditions enabling solubility/permeability studies by stakeholders. Reproducible tools for HTS mechanocrystallization of APIs and co-crystals will benefit Public Health by creating new or repurposed API compositions exhibiting superior in vivo solubility and bioavailability for a $1 trillion pharmaceuticals market.