Chaita Eliza, Gikas Evagelos, Aligiannis Nektarios
Division of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, Athens, 15771, Greece.
Division of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, Athens, 15771, Greece.
Phytochem Anal. 2017 Mar;28(2):125-131. doi: 10.1002/pca.2670. Epub 2017 Feb 2.
In drug discovery, bioassay-guided isolation is a well-established procedure, and still the basic approach for the discovery of natural products with desired biological properties. However, in these procedures, the most laborious and time-consuming step is the isolation of the bioactive constituents. A prior identification of the compounds that contribute to the demonstrated activity of the fractions would enable the selection of proper chromatographic techniques and lead to targeted isolation.
The development of an integrated HPTLC-based methodology for the rapid tracing of the bioactive compounds during bioassay-guided processes, using multivariate statistics. Materials and Methods - The methanol extract of Morus alba was fractionated employing CPC. Subsequently, fractions were assayed for tyrosinase inhibition and analyzed with HPTLC. PLS-R algorithm was performed in order to correlate the analytical data with the biological response of the fractions and identify the compounds with the highest contribution. Two methodologies were developed for the generation of the dataset; one based on manual peak picking and the second based on chromatogram binning. Results and Discussion - Both methodologies afforded comparable results and were able to trace the bioactive constituents (e.g. oxyresveratrol, trans-dihydromorin, 2,4,3'-trihydroxydihydrostilbene). The suggested compounds were compared in terms of R values and UV spectra with compounds isolated from M. alba using typical bioassay-guided process.
Chemometric tools supported the development of a novel HPTLC-based methodology for the tracing of tyrosinase inhibitors in M. alba extract. All steps of the experimental procedure implemented techniques that afford essential key elements for application in high-throughput screening procedures for drug discovery purposes. Copyright © 2017 John Wiley & Sons, Ltd.
在药物研发中，生物测定导向的分离是一种成熟的方法，并且仍然是发现具有所需生物学特性的天然产物的基本途径。然而，在这些方法中，最费力且耗时的步骤是生物活性成分的分离。预先鉴定对各馏分所显示活性有贡献的化合物，将有助于选择合适的色谱技术并实现靶向分离。
开发一种基于高效薄层色谱（HPTLC）的综合方法，利用多元统计技术在生物测定导向过程中快速追踪生物活性化合物。材料与方法 - 采用逆流色谱法（CPC）对桑白皮甲醇提取物进行分离。随后，对各馏分进行酪氨酸酶抑制活性测定，并采用HPTLC进行分析。执行偏最小二乘回归（PLS - R）算法，以便将分析数据与各馏分的生物学响应相关联，并鉴定贡献最大的化合物。开发了两种方法来生成数据集；一种基于手动峰挑选，另一种基于色谱峰分箱。结果与讨论 - 两种方法都得到了可比的结果，并且能够追踪生物活性成分（例如氧化白藜芦醇、反式二氢桑色素、2,4,3'-三羟基二氢茋）。将所建议的化合物的Rf值和紫外光谱与使用典型生物测定导向方法从桑白皮中分离得到的化合物进行了比较。
化学计量学工具支持开发一种基于HPTLC的新方法，用于追踪桑白皮提取物中的酪氨酸酶抑制剂。实验过程的所有步骤都采用了可为药物发现目的的高通量筛选程序应用提供关键要素的技术。版权所有© 2017约翰威立父子有限公司。
