Departamento de Ciências Farmacêuticas, Universidade Federal de Santa Catarina, Campus Universitário-Trindade, Florianópolis, SC, 88040-900, Brazil.
Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Campus Universitário-Trindade, Florianópolis, SC, 88040-900, Brazil.
J Chromatogr A. 2019 Nov 8;1605:460362. doi: 10.1016/j.chroma.2019.460362. Epub 2019 Jul 12.
Sesquiterpene lactones (SL) are commonly found in Asteraceae and present a promising anti-inflammatory activity. Previously described in Lepidaploa genus, glaucolide B has never been investigated for its anti-inflammatory potential. This study aimed to establish an efficient process for the extraction of glaucolide B (1) from Lepidaploa chamissonis leaves and to develop a simple and fast method for its purification by using centrifugal partition chromatography (CPC), as well as to investigate in vitro the anti-inflammatory effects of glaucolide B. Thus, an optimized washing extractive process performed on L. chamissonis leaves allowed to obtain a SL enriched extract (4.11 g). After a successful defatting pretreatment of the crude extract, the glaucolide B enriched ethyl acetate portion (2.00 g) was fractionated by CPC affording, in a single-step isolation, compound 1 (1.04 g) in great yield (25%) and purity (97%). Cytotoxicity effect of 1 on RAW 264.7 macrophages was determined by using MTT assay, revealing a CC of 14.11 μM. Compound 1 at 1, 3 and 10 μM inhibited the nitrite/nitrate (NO) metabolites production and the pro-inflammatory interleukin 6 (IL-6) secretion on lipopolysaccharide-stimulated RAW 264.7 cells. The extractive process used turned to be selective for SL and CPC technique proved a simple and effective tool for the isolation of 1 within few hours. Isolated for the first time from L. chamissonis leaves, glaucolide B presented a significant inhibitory effect on both NO and IL-6 secretion under non-toxic concentrations.
倍半萜内酯（SL）普遍存在于菊科中，具有有前景的抗炎活性。在 Lepidaploa 属中已有描述，从未研究过 glaucolide B 的抗炎潜力。本研究旨在建立从 Lepidaploa chamissonis 叶中提取 glaucolide B（1）的有效方法，并开发一种使用离心分配色谱（CPC）进行其简单快速纯化的方法，以及研究体外 glaucolide B 的抗炎作用。因此，对 L. chamissonis 叶片进行优化的洗涤提取过程可获得富含 SL 的提取物（4.11g）。在对粗提取物进行成功脱脂预处理后，用 CPC 对富含 glaucolide B 的乙酸乙酯部分（2.00g）进行分步分离，在单次分离中获得化合物 1（1.04g），收率高（25%），纯度（97%）。通过 MTT 测定法测定了 1 对 RAW 264.7 巨噬细胞的细胞毒性作用，发现 CC 为 14.11μM。化合物 1 在 1、3 和 10μM 浓度下抑制了脂多糖刺激的 RAW 264.7 细胞中硝酸盐/亚硝酸盐（NO）代谢产物的产生和促炎白细胞介素 6（IL-6）的分泌。所使用的提取过程对 SL 具有选择性，CPC 技术证明了在数小时内分离 1 的简单有效工具。首次从 L. chamissonis 叶片中分离出的 glaucolide B 在非毒性浓度下对 NO 和 IL-6 分泌均具有显著抑制作用。
