Chrysanthemum boreale Makino (C. boreale) is widely distributed in Asian countries, and has traditionally been used to treat various inflammatory diseases including bronchitis. In this study, we aimed to isolate biologically active compounds from leaves and stems of C. boreale. Chemical components were purified by column chromatograpy and recyclic HPLC, and characterized from their spectral data (IR, MS, NMR). Biological activity experiments were conducted for Farnesyl-protein transferase (FPTase) activity, apoptosis and nitirc oxide (NO) release. As a results, three sesquiterpene lactones were isolated. Compound 1 (4-methoxy-8-O-acetyl-10-hydroxy-2,11(13)-guaiadiene-12,6-olide) showed strong cytotoxic activities having an average growth inhibition of 50% (GI50) value of 1.89 ㎍/㎖against human colon adenocarcinoma cells. Compound 1 also showed a low half maximal inhibitory concentration (IC50) value of 10 ㎍/㎖ for NO release. In the caspase 3 activity, compound 1 and compound 2 (8-O-(2-carbonyl-2-butyl)-3,10-dihydroxy-4,11(13) -guaiadiene-12,6-olide) exhibited 94% and 90% apoptosis inhibition activity, respectively. Compound 3 (4,8-O-diacetyl -10-hydroxy-2(3),11(13)-guaiadiene-12,6-olide) showed a strong inhibitory effect on FPTase activity with 90% inhibitory activity at a concentration of 100 ㎍/㎖. These results clearly show the presence of lactone compounds in the leaves and stems, which may partially contribute to the pharmacological activity of C. boreale.

Chrysanthemum boreale Makino is widely distributed in Korea, China, Japan and Southeast Asian countries. C. boreale is one of the herbs used for treating various inflammatory diseases in oriental medicine. The present study was conducted to identify biologically active compounds from the leaves and stems of C. boreale. We isolated two sesquiterpene sactones from the leaves and stems of C. boreale using silica gel column chromatography and recyclic high perfomance liquid chromatography. The lactones were characterized by their spectroscopic data (NMR, IR, MASS). These compounds were subjected to Farnesyl Protein Transferase (FPTase) inhibition, Nitric Oxide (NO) release inhibition and apoptosis inhibition. The structur of the following isolated compound were elucidated 8,10-o-Acetyl-2-methoxy-10-hydroxy-3,11(13)-guaiadiene-12,6-olide and 4,10-dihydroxy -8-o-Acetyl-2,11(13)-guaiadiene-12,6-olide. In the NO release inhibition assay, compound 2 showed strong activities, with an IC50 value of 7 μg/mL, whereas compound 1 did not exhibit significant activity with an IC50 value of over 14 μg/mL against murine macrophage.

Acyltransferase (AT) catalyzes the transfer of an acyl moiety from acyl-coenzyme A (acyl-CoA) to an acceptor. ATs play important roles in the maintenance of homeostasis in the human body and have been linked to various diseases; therefore, several ATs have been proposed as potential targets for the treatment or prevention of such diseases. The AT family includes acyl-CoA:cholesterol AT (ACAT), diacylglycerol AT, and monoacylglycerol AT for the metabolism of lipids. Furthermore, recent molecular biological studies revealed the existence of their isozymes with distinct functions in the body. ACAT plays a critical role in the formation of cholesteryl esters from cholesterol and fatty acids, and is a potential target for treating hypercholesterolemia. During an experiment designed to discover biologically active compounds from herbal medicines, we isolated two known guaianolide sesquiterpene lactones from Chrysanthemum boreale Makino (Compositae). The lactones were characterized from their spectroscopic data (NMR, IR, MASS). These compounds were subjected to ACAT inhibition assay. Here, we report the isolation and structural elucidation of the compounds 8-o-acetyl-2-methoxy-10-hydroxy-3,11(13)-guaiadiene-12,6-olide and 8-acetyl-3,10-hydroxy-4(15),11(13)-guaiadiene-12,6-olide. In the ACAT inhibition assay, compound 1 showed strong inhibitory activity, with an IC50 value 45 μg/mL, whereas compound 2 did not exhibit significant inhibitory activity with an over 100 μg/mL.

본 연구는 수증기 증류법을 이용하여 추출한 산국화 유래 hydrosol의 피부각질형성세포(HaCaTs) 증식 및 이주 유도 활성 평가를 통해 피부재상피화 및 상처치유 활성을 확인하였다. 산국화 hydrosol은 HaCaTs의 증식과 이주를 농도 의존적으로 유도하였으며, 특히 1 μg/mL에서 음성 대조군(control)에 비해 143.71 ± 3.37%의 증식과, 139.98 ± 5.72%의 이주 유도 활성을 나타내었다. 또한, 산국화 hydrosol은 extracellular signal-regulated kinase 1/2 (Erk 1/2)와 serine/threonine-specific protein kinase (Akt)의 인산화를 유 의하게 증가시켰을 뿐만 아니라 collagen sprout out growth를 농도 의존적으로 유도하였다. 이러한 결과들을 통해서 산국화 hydrosol은 정상적인 재상피화 과정과 상처치유 등의 활성이 있음을 확인하였고, 향후 화장품 소재로서 응용가능성을 검증하였다.

The flowers of Chrysanthemum boreale Makino were extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with n-hexane, EtOAc, n-BuOH and H2O. Two compounds from the n-hexane fraction and one glucoside from the n-BuOH fraction were isolated through the repeated silica gel and ODS column chromatographies. From the result of physico-chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as β-sitosterol (1), phytol (2) and zingerone 4-O-β-D-glucopyranoside (3). Compounds 2 and 3 were isolated for the first time from this plant.

천연물로부터 항산화 활성물질을 목적으로 DPPH 자유 라디칼 소거활성 검색법을 이용하여 검색을 실사하고, 유의한 활성을 나타낸 산국 메탄올 추출물로부터 2종의 화합물을 분리하고 기기분석을 이용하여 화합물 1과 2의 구조를 apigenin과 acacetin-7-O-rutinoside (Linarin)으로 동정하였다. 이들 화합물은 각각 13.3 및 42.1μg의 농도에서 DPPH 라디칼을 50% 소거하는 활성을 나타내었으며, 화합물 1은 양성 대조양물인 L-ascorbic acid (RC50 = 13.1μg)에 비하여 강한 활성을 나타내었다.