Department of Drug Design and Pharmacology, Faculty of Health and Medical Science, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
Department of Drug Design and Pharmacology, Faculty of Health and Medical Science, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
J Chromatogr A. 2018 Jun 29;1556:55-63. doi: 10.1016/j.chroma.2018.04.041. Epub 2018 Apr 21.
In this paper, quadruple high-resolution α-glucosidase/α-amylase/PTP1B/radical scavenging profiling combined with HPLC-HRMS-SPE-NMR were used for studying the polypharmacological properties of crude root bark extract of Morus alba L. This species is used as an anti-diabetic principle in many traditional treatment systems around the world, and the crude ethyl acetate extract of M. alba root bark was found to inhibit α-glucosidase, α-amylase and protein-tyrosine phosphatase 1B (PTP1B) with IC values of 1.70 ± 0.72, 5.16 ± 0.69, and 5.07 ± 0.68 μg/mL as well as showing radical scavenging activity equaling a TEAC value of (3.82 ± 0.14) × 10 mM per gram extract. Subsequent investigation of the crude extract using quadruple high-resolution α-glucosidase/α-amylase/PTP1B/radical scavenging profiling provided a quadruple biochromatogram that allowed direct correlation of the HPLC peaks with one or more of the tested bioactivities. This was used to target subsequent HPLC-HRMS-SPE-NMR analysis towards peaks representing bioactive analytes, and led to identification of a new Diels-Alder adduct named Moracenin E as well as a series of Diels-Alder adducts and isoprenylated flavonoids as potent α-glucosidase and α-amylase inhibitors with IC values in the range of 0.60-27.15 μM and 1.22-69.38 μM, respectively. In addition, these compounds and two 2-arylbenzofurans were found to be potent PTP1B inhibitors with IC values ranging from 4.04 to 21.67 μM. The high-resolution radical scavenging profile also revealed that almost all of the compounds possess radical scavenging activity. In conclusion the quadruple high-resolution profiling method presented here allowed a detailed profiling of individual constituents in crude root bark extract of M. alba, and the method provides a general tool for detailed mapping of bioactive constituents in polypharmacological herbal remedies.
本文采用四重高分辨率α-葡萄糖苷酶/α-淀粉酶/蛋白酪氨酸磷酸酶 1B/自由基清除谱分析结合高效液相色谱-高分辨质谱-固相萃取-核磁共振波谱法，研究了桑白皮的粗提取物的多药效性质。该物种被许多世界各地的传统治疗系统用作抗糖尿病的原理，并且发现桑白皮根的粗乙酸乙酯提取物抑制α-葡萄糖苷酶、α-淀粉酶和蛋白酪氨酸磷酸酶 1B（PTP1B）的 IC 值分别为 1.70 ± 0.72、5.16 ± 0.69 和 5.07 ± 0.68μg/mL，并且显示自由基清除活性相当于每克提取物的 TEAC 值为（3.82 ± 0.14）×10mM。随后使用四重高分辨率α-葡萄糖苷酶/α-淀粉酶/PTP1B/自由基清除谱对粗提取物进行研究，提供了四重生物色谱图，可将 HPLC 峰与一种或多种测试的生物活性直接相关联。这用于将随后的 HPLC-HRMS-SPE-NMR 分析针对代表生物活性分析物的峰，导致鉴定出一种新的 Diels-Alder 加合物，命名为 Moracenin E，以及一系列 Diels-Alder 加合物和异戊烯基化黄酮类化合物，它们是有效的α-葡萄糖苷酶和α-淀粉酶抑制剂，IC 值范围为 0.60-27.15μM 和 1.22-69.38μM，分别。此外，这些化合物和两种 2-芳基苯并呋喃被发现是有效的 PTP1B 抑制剂，IC 值范围为 4.04-21.67μM。高分辨率自由基清除谱还表明，几乎所有化合物都具有自由基清除活性。总之，本文提出的四重高分辨率分析方法允许对桑白皮粗提取物中的单个成分进行详细分析，该方法为多药效草药制剂中生物活性成分的详细映射提供了一种通用工具。
