HIGH SPEED SFC PURIFICATION OF COMBI-CHEM LIBRARIES

这项拟议的研究应导致开发和表征的半制备规模的超临界流体色谱（SFC）优化纯化组合化学库，并提供高达2个数量级的增加，在传统的制备HPLC的吞吐量。 这将使得纯化整个combi-chem文库在经济上是可行的，从而导致更少的假阳性和假阴性筛选结果。 SFC纯化比使用平行柱的制备型HPLC的生产率高得多。 SFC分离所需的时间仅为HPLC的1/10左右，而且通过无需冻干，可显著加速溶剂从分离馏分中的去除，并消除大量混合水/有机废物的产生和处理。 对原型系统的初步测试表明，该系统每天能够净化200个样品，溶质量为1-100 mg，回收率和纯度高于95%。气/液分离器设计允许收集每个样品的多个级分，从而不仅能够回收反应产物，而且能够回收高度纯化状态的稀缺的未反应的起始材料。 本研究的重点是优化和表征系统的药物发现化合物的多样性，并证明该系统与机器人馏分收集器集成的可行性。拟议的商业应用：所提出的半制备型SFC系统，当与实验室机器人系统完全自动化时，应消除药物发现过程中的主要瓶颈，并允许以当前HPLC技术在经济上不可行的规模纯化整个combi-chem文库。这将导致组合化学文库更有可能产生与已知纯化学实体相关的阳性命中。

This proposed research should lead to development and characterization of a semi-preparative scale Supercritical Fluid chromatograph (SFC) optimized for purifying combi-chem libraries and offering up to 2 orders of magnitude increase in throughput over traditional preparative HPLC. This would make it economically feasible to purify entire combi-chem libraries, leading to fewer false positive and false negative screening results. SFC purification is far more productive than preparative HPLC with parallel columns. Not only do SFC separations take approximately 1/10th the time of HPLC, but SFC dramatically accelerates solvent removal from separated fractions by removing the need for lyophilization, and eliminates generation and disposal of vast amounts of mixed aqueous/organic waste. Preliminary testing of a prototype system show that the system is capable of purifying 200 samples per day with solute quantities of 1-100 mg, with better than 95% recovery and purity. The gas/liquid separator design allows for collecting multiple fractions per sample, enabling recovery of not only reaction products but also of scarce unreacted starting materials in a highly purified state. The focus of this research is to optimize and characterize the system for a diversity of drug discovery compounds, and demonstrate feasibility of integrating this system with a robotic fraction collector. PROPOSED COMMERCIAL APPLICATIONS: The proposed Semi-prep SFC system, when fully automated with a laboratory robotic system, should eliminate a major bottleneck in the drug discovery process and allow for purification of entire combi-chem libraries on a scale not economically feasible with current HPLC technology. This will result in combi-chem libraries far more likely to yield positive hits associated with a known pure chemical entity.