Comparative Study of Chromatographic Medium-Associated Mass and Potential Antitumor Activity Loss with Bioactive Extracts
Comparative Study of Chromatographic Medium-Associated Mass and Potential Antitumor Activity Loss with Bioactive Extracts
复制标题
DOI：
10.1021/np300858c
复制
发表时间：
2013-04-01
影响因子：
5.1
通讯作者：
Nagle, Dale G.
中科院分区：
文献类型：
--
作者：
Datta, Sandipan;Zhou, Yu-Dong;Nagle, Dale G.
天然产物药物发现程序通常依赖于使用二氧化硅（Si）凝胶、反相介质或尺寸排阻树脂（例如，RP-C-18，Sephadex LH-20）用于化合物纯化。合成聚合物基吸附剂Diaion HP 2 OSS（交联聚苯乙烯基质）被用作制备纯化天然产物库的替代品。为了评价色谱介质对分离具有生物活性但色谱不稳定的天然产物的影响，对Diaion HP 2 OSS与正相吸附剂进行了并行评价，以确定提取物成分的不可逆结合及其对生物活性的影响。选择一系列化学上不同的富含天然产物的提取物作为测试组，并采用缺氧诱导因子-1（HIF-1）的基于细胞的报告基因测定来监测生物活性的潜在变化。硅胶导致10种浸提液中有3种发生显著的不可逆结合。姜黄、三白草和柑橘提取物通过硅胶洗脱后显示出降低的HIF-1抑制活性。用EtOAc对HP 2 OSS进行额外的非极性柱洗涤保留了活性提取物的相当大的生物活性。一般情况下，硅凝胶产生的生物活性损失最大。然而，HP 2 OSS洗脱显著降低了某些提取物（例如，Asimina triloba）。
Natural product drug discovery programs often rely on the use of silica (Si) gel, reversed-phase media, or size-exclusion resins (e.g., RP-C-18, Sephadex LH-20) for compound purification. The synthetic polymer-based sorbent Diaion HP2OSS (cross-linked polystyrene matrix) is used as an alternative to prepare purified natural product libraries. To evaluate the impact of chromatographic media on the isolation of biologically active, yet chromatographically unstable natural products, Diaion HP2OSS was evaluated side-by-side with normal-phase sorbents for irreversible binding of extract constituents and their effects on bioactivity. An array of chemically diverse natural product-rich extracts was selected as a test panel, and a cell-based reporter assay for hypoxia-inducible factor-1 (HIF-1) was employed to monitor potential change(s) in bioactivity. Silica gel caused significant irreversible binding of three out of 10 extracts. Curcuma longa, Saururus cerriuus, and Citrus reticulata extracts showed decreased HIF-1 inhibitory activity after elution through Si gel. An additional nonpolar column wash of HP2OSS with Et0Ac retained considerable bioactivities of active extracts. In general, Si gel produced the greatest loss of bioactivity. However, HP2OSS elution reduced significantly HIF-1 inhibitory activity of certain extracts (e.g., Asimina triloba).