A microED pipeline for the pharmaceutical and biotechnology industry

适用于制药和生物技术行业的 microED 管道

• 批准号: 10402253
• 负责人: Jessica Fox Bruhn
• 金额: $ 68.37万
• 依托单位: NANOIMAGING SERVICES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10402253
• 关键词: 3-Dimensional; Acetaminophen; Address; Algorithms; Automation; Biological Products; Biotechnology; Biotin; Client; Clinic; Collection; Commercial Sectors; Complex; Computer software; Crystallization; Data; Data Collection; Data Set; Development; Drug Costs; Drug Design; Electron Microscope; Ensure; Event; Feedback; Goals; Handedness; Health; Hour; Human; Image; Industrialization; Industry; Knowledge; Maps; Methods; Micro Electron Diffraction; Microscope; Modeling; Molecular; Molecular Target; Natural Products; Online Systems; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Preparation; Procedures; Process; Progesterone; Property; Proteins; Reproducibility; Research; Resolution; Robot; Rotation; Sampling; Series; Services; Specificity; Specimen; Speed; Structure; System; Techniques; Testing; Time; Work; X-Ray Crystallography; base; chemotherapy; chiral molecule; computerized data processing; cost; crystallinity; data acquisition; data analysis pipeline; data quality; data reduction; drug development; drug discovery; electron diffraction; flexibility; improved; improved outcome; interest; microscopic imaging; nanoimaging; novel; novel strategies; programs; screening; small molecule; statistics; targeted treatment; three dimensional structure; transmission process

Project Summary/Abstract Microcrystal electron diffraction (microED) is an emerging structure determination technique of high interest to the pharmaceutical and biotechnology industries. MicroED can rapidly determine atomic-resolution structures from microcrystals with minimal sample requirements. The goal of this Phase II proposal is to develop and deliver robust service offerings for microED structure determination to the commercial sector. Improvements to critical steps in this workflow will enable analysis for any suitable sample type while offering a level of efficiency that meets the requirements of Structure-Based Drug Design (SBDD) and medicinal chemistry programs. The permissive sample requirements, minimal time requirements, and high-quality structures generated make microED an attractive structure determination technique with broad applications for small molecules, peptides, natural products, proteins and protein-drug complexes. An efficient microED pipeline will help to reduce the cost of drug development and reduce the time it takes a drug to make it into the clinic. MicroED uses a transmission electron microscope (TEM) to collect diffraction data from small crystalline samples. The steps required include transferring crystalline samples to TEM grids; collecting a series of continuous-rotation, electron-diffraction datasets from single crystals; processing the diffraction data and refining the atomic model. An initial development phase demonstrated collection of excellent quality diffraction datasets for more than 20 samples, and the ability to deliver atomic resolution maps suitable for addressing the needs of our pharmaceutical clients who took part in alpha and beta testing. New methods will be developed, and existing approaches optimized to improve every step of this pipeline, substantially increase throughput, and improve final maps and models. Specific aims include: (1) Optimizing TEM grid preparation to ensure the transfer of a sufficient number of high-quality target microcrystals onto the grid substrate without damaging them or altering their properties. (2) Developing an automated, high-throughput data collection system capable of unattended, overnight data acquisition for multiple different samples, and of providing information to inform data scaling and determining handedness of chiral small molecules. (3) Automating and improving data processing, phasing and model refinement including real-time data reduction, brute force on-the-fly phasing, and investigating a wide range of new programs and methods. In parallel with the Phase II research plan, a full commercial service offering spanning the complete range of microED capabilities will be developed.

项目总结/摘要 微晶电子衍射（microED）是一种新兴的结构测定技术， 制药和生物技术行业。MicroED可以快速确定原子分辨率的结构 用最少的样品要求从微晶中提取。第二阶段提案的目标是开发和交付 为商业部门的microED结构确定提供强大的服务。关键改进 该工作流程中的步骤将能够分析任何合适的样品类型，同时提供一定的效率水平， 符合基于结构的药物设计（SBDD）和药物化学计划的要求。的 宽松的样品要求，最短的时间要求，以及高质量的结构生成， microED是一种有吸引力的结构测定技术，广泛应用于小分子，肽， 天然产物、蛋白质和蛋白质-药物复合物。高效的microED管道将有助于降低成本 减少药物进入临床的时间。 MicroED使用透射电子显微镜（TEM）收集来自小晶体的衍射数据， 样品所需的步骤包括将晶体样品转移到TEM网格上;收集一系列样品。 单晶体的连续旋转电子衍射数据集;处理衍射数据并细化 原子模型。最初的开发阶段展示了高质量衍射数据集的收集 超过20个样品，并能够提供原子分辨率地图适合解决的需求， 我们的制药客户参与了alpha和beta测试。新的方法将被开发，现有的 优化的方法，以改善这一管道的每一步，大大提高吞吐量，并提高最终 地图和模型。具体目标包括：（1）优化TEM网格制备，保证传输足够的透射率; 将大量高质量的目标微晶粘附到网格衬底上，而不损坏它们或改变它们的 特性. (2)开发一种自动化、高通量的数据收集系统， 多个不同样品的过夜数据采集，并提供信息以告知数据缩放和 确定手性小分子的手性。(3)自动化和改进数据处理、分阶段和 模型细化，包括实时数据减少，蛮力对飞行阶段，并调查了广泛的 一系列新的方案和方法。 在第二阶段研究计划的同时，一个完整的商业服务提供，涵盖了整个范围， 将开发微型电子设备。

项目成果

Small Molecule Microcrystal Electron Diffraction for the Pharmaceutical Industry-Lessons Learned From Examining Over Fifty Samples.

• DOI: 10.3389/fmolb.2021.648603
• 发表时间: 2021
• 期刊: Frontiers in molecular biosciences
• 影响因子: 5
• 作者: Bruhn JF;Scapin G;Cheng A;Mercado BQ;Waterman DG;Ganesh T;Dallakyan S;Read BN;Nieusma T;Lucier KW;Mayer ML;Chiang NJ;Poweleit N;McGilvray PT;Wilson TS;Mashore M;Hennessy C;Thomson S;Wang B;Potter CS;Carragher B
• 通讯作者: Carragher B

Absolute Configuration Determination of Chiral API Molecules by MicroED Analysis of Cocrystal Powders Formed Based on Cocrystal Propensity Prediction Calculations.

通过基于共晶倾向预测计算形成的共晶粉末的 MicroED 分析来确定手性 API 分子的绝对构型。

• DOI: 10.1002/chem.202203970
• 发表时间: 2023
• 期刊: Chemistry (Weinheim an der Bergstrasse, Germany)
• 作者: Shah,HarshS;Yuan,Jiuchuang;Xie,Tian;Yang,Zhuocen;Chang,Chao;Greenwell,Chandler;Zeng,Qun;Sun,GuangXu;Read,BrandonN;Wilson,TimothyS;Valle,HenryU;Kuang,Shanming;Wang,Jian;Sekharan,Sivakumar;Bruhn,JessicaF
• 通讯作者: Bruhn,JessicaF

From Powders to Single Crystals: A Crystallographer's Toolbox for Small-Molecule Structure Determination.

• DOI: 10.1021/acs.molpharmaceut.2c00020
• 发表时间: 2022-07-04
• 期刊: MOLECULAR PHARMACEUTICS
• 影响因子: 4.9
• 作者: Newman, Justin A.;Iuzzolino, Luca;Tan, Melissa;Orth, Peter;Bruhn, Jessica;Lee, Alfred Y.
• 通讯作者: Lee, Alfred Y.

Absolute configuration determination of pharmaceutical crystalline powders by MicroED via chiral salt formation.

• DOI: 10.1039/d2cc00221c
• 发表时间: 2022-04-12
• 期刊: CHEMICAL COMMUNICATIONS
• 影响因子: 4.9
• 作者: Wang, Bo;Bruhn, Jessica F.;Weldeab, Asmerom;Wilson, Timothy S.;McGilvray, Philip T.;Mashore, Michael;Song, Qiong;Scapin, Giovanna;Lin, Yiqing
• 通讯作者: Lin, Yiqing