Spiral Countercurrent Chromatography for High-throughput Natural Product Purifica

描述(申请人提供)：逆流色谱(CCC)的最新进展创造了新的螺旋设计的分离柱，可以保持所有溶剂系统的非常高的固定相，因此现在任何大小和水溶性的分子都可以在高速逆流色谱中成功分离。与任何类型的层析相比，螺旋CCC具有固有的多功能性。新型螺旋管支撑转子的初步研究成功地提纯了甜菜中主要的水溶性环丙烷苷B，该糖苷使用正丁醇溶剂体系。由于线圈体积很小(~100毫升)，固定相为50%(~50毫升)，一次可以有效地分离1.2g的植物粗提物，这是令人印象深刻的 具有较高的目标化合物回收率。此外，还用类似的方法一步分离得到了活性环烯醚萜糖苷haragoside haragoside(Haragoside)。第一阶段的第一个具体目标是设计和制造一种制备性规模的螺旋CCC仪器，该仪器将用于快速分离具有高通量能力的复杂的水溶性天然产物。这些研究将包括测试更大体积的分离转子，然后将其安装到放大的行星离心机中。商业化前的原型测试将与密苏里大学的顾问威廉·福克博士一起进行。第二个具体目标是初步工艺开发，提纯大量的水溶性提取物，以分别从非洲药用植物沙冬青和普鲁木本中获得每一种山竹皂苷和哈帕皂苷代谢物。将确定一种改进适当的溶剂系统以回收目标化合物的策略。随着化合物的成功提纯，将继续进行第二阶段的研究，与福克博士和密苏里大学合作，在细胞培养和动物模型中表征它们的特性。这项研究的成功成果将是一种商业化仪器--制备型螺旋高速逆流色谱仪，这是一项满足全球研究市场需求的天然产物化学的重要技术。其结果将是提高药物发现的生产率，使替代药物得以特色化，并能够为许多疾病和障碍提供新的治疗方法。最后，丁醇溶剂在螺旋高速逆流色谱中的应用是绿色化学的一个很有希望的例子，因为这些溶剂来自生物燃料。

DESCRIPTION (provided by applicant): Recent advances in countercurrent chromatography (CCC) have created novel spiral-design separation columns that hold a very high stationary phase of all the solvent systems, such that now all molecules of any size and water solubility can be successfully separated in high-speed countercurrent chromatography. With spiral CCC there is inherent versatility, more than any type of chromatography. Preliminary studies with the new spiral tubing support rotor succeeded in purifying Sutherlandioside B, a major water soluble cycloartane glycoside from Lessertia frutescent (Sutherlandia) is using an n- butanol solvent system. With the small volume of the coil (~100 ml) and a stationary phase at 50% (~50 ml), it is impressive that a mass of 1.2 g of crude plant extract could be efficiently fractionated in one run with high recovery of the target compound. In addition, the active iridoid glycoside, harpagoside was isolated from an extract of Harpagophytum procumbens (Devil's Claw) in one step using a similar method. The first specific aim for Phase I am the design and manufacture of a preparative-scale spiral CCC instrument that will serve to rapidly separate complex water-soluble natural products with high- throughput capability. The studies will involve testing of a higher volume separation rotor and then its incorporation into a scaled-up planetary centrifuge. The pre-commercial prototype testing will be performed in conjunction with Dr. William Folk, the consultant from the University of Missouri. The second specific aim is the preliminary process development of purifying significant amounts of water-soluble extracts to obtain each of the sutherlandiosides and harpagoside metabolites from the medicinal African plants, L. frutescens and H. procumbens, respectively. A strategy of modifying a suitable solvent system to recover a target compound will be identified. With the compounds successfully purified, the studies will be continued in phase II to characterize their properties in cell culture and animal models, in collaboration with Dr. Folk and the University of Missouri. The successful outcome of this research will be a commercial instrument, the preparative spiral high-speed countercurrent chromatograph, an important technology for natural products chemistry that meets a demand in the global research market. The result will be increased productivity in drug discovery, enabling alternative medicines to be characterized and new therapies for many diseases and disorders. Finally, the application of butanol solvents so readily in spiral high-speed countercurrent chromatography is a promising example of green chemistry as these solvents are derived from biofuels.