Spiral Countercurrent Chromatography for High-throughput Natural Product Purifica

用于高通量天然产物纯化的螺旋逆流色谱法

• 批准号: 8711698
• 负责人: Martha Knight
• 金额: $ 21.74万
• 依托单位: CC BIOTECH, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8711698
• 关键词: 1-Butanol; Adopted; African; Alternative Medicine; Animal Model; Biological Factors; Businesses; Butanols; Cell Culture Techniques; Chemistry; Chromatography; Claw; Collaborations; Collection; Complex; Crude Extracts; Data Analyses; Detection; Development; Disease; Excision; Fractionation; Glycosides; Laboratory Research; Market Research; Mass Spectrum Analysis; Methods; Missouri; Natural Products Chemistry; One-Step dentin bonding system; Outcomes Research; Performance; Phase; Plant Extracts; Plants; Process; Productivity; Property; Pump; Recovery; Research; Running; Small Business Innovation Research Grant; Solid; Solvents; Speed; System; Technology; Testing; Uncaria procumbens; Universities; Water; aqueous; countercurrent chromatography; design; drug discovery; harpagoside; improved; innovation; instrument; meetings; method development; novel; programs; prototype; public health relevance; response; scale up; small molecule; tool; water solubility

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.

描述（由申请人提供）：逆流色谱（CCC）的最新进展已经产生了新颖的螺旋设计分离柱，其保持所有溶剂系统的非常高的固定相，使得现在可以在高速逆流色谱中成功地分离任何尺寸和水溶性的所有分子。螺旋CCC具有固有的多功能性，超过任何类型的色谱。利用新的螺旋管支持转子的初步研究成功地纯化了Sutherlandioside B，Sutherlandia是一种主要的水溶性环阿屯烷糖苷，来自Lessertia frutescent（Sutherlandia），使用正丁醇溶剂系统。在小体积的螺旋管（~100 ml）和50%（~50 ml）的固定相的情况下，令人印象深刻的是，在一次运行中可以有效地分级分离1.2 g的粗植物提取物 目标化合物的回收率高。此外，活性环烯醚萜苷，哈巴俄苷是使用类似的方法在一个步骤中从Harpagophytum procumbens（Devil's Claw）的提取物中分离的。第一阶段的第一个具体目标是设计和制造一种分离规模的螺旋CCC仪器，该仪器将用于以高通量能力快速分离复杂的水溶性天然产物。这些研究将包括测试一个容量更大的分离转子，然后将其纳入一个按比例放大的行星式离心机。将与来自密苏里州大学的顾问William Folk博士一起进行商业前原型测试。第二个具体目标是初步开发纯化大量水溶性提取物以从药用非洲植物L. frutescens和H.平卧，分别。将确定修改合适的溶剂系统以回收目标化合物的策略。随着化合物的成功纯化，将与Folk博士和密苏里州大学合作，在第二阶段继续进行研究，以表征其在细胞培养和动物模型中的特性。这项研究的成功成果将是一个商业化的仪器，制备螺旋高速逆流色谱仪，一个重要的技术，天然产物化学，满足全球研究市场的需求。其结果将是提高药物发现的生产力，使替代药物的特点和新的疗法，许多疾病和病症。最后，丁醇溶剂如此容易地应用于螺旋高速逆流色谱是绿色化学的一个有前途的例子，因为这些溶剂来自生物燃料。