분홍바늘꽃, 편백 종자 및 팥배나무 추출물의 기능성 화장품 소재 개발 연구

Abstract

In this study, we investigated anti-oxidative, anti-bacterial, anti-inflammatory and whitening constituents from aerial parts of Epilobium angustifolium L., seeds of Chamaecyparis obusa (Siebold. et Zucc.) Endl., leaves and branches of Sorbus alnifolia (Sieb. et Zucc.) K. Koch. Dried plant samples were extracted with 70% or 50% aqueous ethanol, and crude extracts were subjected to solvent fractionation according to polarity. The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including NMR and HR-ESI-MS spectra, as well as comparison of the data to the literature values. Twenty one constituents were isolated from the extract of Epilobium angustifolium L. aerial parts; n-dodecane (1), lauric acid (2), ethyl oleate (3), 1-monoolein (4), linoleic acid (5), coriolic acid (6), ethyl linolenate (7), α-linolenic acid (8), 1-linolenoyl glycerol (9), 15-isopimaren-3β,8β-diol (10), α-amyrin (11), β-amyrin (12), ursolic acid (13), oleanolic acid (14), maslinic acid (15), corosolic acid (16), β-sitosterol (17), 5-desmethylsinensetin (18), p-coumaric acid (19), caffeic acid (20), (S)-danshensu caffeic anhydride (21). As far as we know compound 21 was identified as a novel compound found in nature. Upon the anti-oxidative studies by DPPH and ABTS+ radicals, potent radical scavenging activities were observed in n-butanol (BuOH) fraction and extract. The extract, n-hexane (Hex) and ethyl acetate (EtOAc) fractions showed anti-bacterial activities against staphylococcus epidermidis, Streptococcus mutans and Cutibacterium acnes. Also, in the anti-inflammatory tests using RAW264.7 mouse macrophages, the n-Hex and EtOAc fractions inhibited the production of nitric oxide (NO) without causing cell toxicity. In addition, the n-Hex and EtOAc fractions reduced production of prostaglandin (PG) E2 and pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β. In the whitening tests using B16F10 melanoma cells, the n-Hex and EtOAc fractions concentration-dependently inhibited cellular melanogenesis and intracellular tyrosinase activity without causing cell toxicity. Moreover, isolated compound 21 from EtOAc fraction showed potent free radical scavenging activities, excellent anti-inflammatory activities, and anti-melanogenesis inhibition effects. Forty one constituents were isolated from the extract of Chamaecyparis obusa (Siebold. et Zucc.) Endl. seeds; (R)-p-menth-1-en-4,7-diol (1), (1R,2R,4R)-p-menthane-1,2,4-triol (2), α-eudesmol (3), γ-eudesmol (4), β-eudesmol (5), oplodiol (6), 4-eudesmene-1β,11-diol (7), ent-4(15)-eudes-men-1α,11-diol (8), 3-eudesmene-1β,11-diol (9), hinokiic acid (10), 1α-hydroxy-hinokione (11), 12-methoxy-8,11,3-abietatriene-7β,11-diol-3-o-ne (12), hinokione (13), 1,2-dehydrohinokione (14), 1α-3β-dihydroxytota-rol (15), hinokiol (16), isohinokiol (17), sugiol (18), ferruginol (19), cryptojaponol (20), 7α,11-dihydroxy-12-methoxy-8,11,13-abietatriene (21), 7β-hydroxydeoxocryptojaponol (22), 6,7-dehydrodeoxocryptojaponol (23), trans-communic acid (24), chamaecydin (25), α-linolenic acid methyl ester (26), α-linolenic acid (27), deoxypodophyllotoxin (28), yatein (29), hinokinin (30), savinin (31), haplomyrfolin (32), sesamin (33), aromadendrin (34), taxifolin (35), taxifolin-3-O-β-D-xylopyranoside (36), taxifolin-3-O-α-L-rhamnopyranoside (37), apigenin (38), scutellarein (39), quercetin (40), quercitrin (41). As far as we know the compound 11 was identified as a novel compound found in nature. Upon the anti-oxidative studies by DPPH and ABTS+ radicals, potent radical scavenging activities were observed in extract, EtOAc, n-BuOH fractions and isolated compound 32. The extract, n-Hex and EtOAc fractions showed anti-bacterial activities against S. epidermidis, S. mutans and C. acnes. Also, in the anti-inflammatory tests using RAW264.7 mouse macrophages, the extract and n-Hex, EtOAc fractions inhibited the production of nitric oxide (NO) without causing cell toxicity. In addition, the extract, n-Hex and EtOAc fractions reduced production of pro-inflammatory cytokine TNF-α. In the whitening tests using mushroom tyrosinase, the n-Hex, EtOAc fractions and isolated compounds 18, 19, 21, 24 concentration-dependently inhibited tyrosinase activity. On the other hand, among the isolates, compounds 3, 10-15, 19, 21, 22, 24 showed potent anti-bacterial activities against S.epidermidis and C. acnes. Also, isolated compounds 11, 13-15, 17, 20, 22, 29, 32 showed strong anti-inflammatory activities. Twelve phytochemicals were isolated from the extract of Sorbus alnifolia leaves; ursolic acid (1), oleanolic acid (2), α-amyrin (3), corosolic acid (4), (E)-phytol (5), α-linolenic acid (6), α-linolenic acid methyl ester (7), 1-linolenoyl glycerol (8), afzelin (9), kaempferitrin (10), kaempferol 3-O-β-D-xylopyranosyl-(1→2)-α-L-rhamnopyranosyl-7-O-α-L-rhamnopyranos-ide (11), kaempferol 3-O-[deoxy-ribo-hexos-3-ulosyl-(1→2)-α-L-rham-nopyranosyl]-7-O-α-L-rhamnopyranoside (12). As far as we know, the compound 12 was identified as a novel compound found in nature. On the anti-oxidative tests, the EtOAc, n-BuOH fractions and isolated compounds 9, 10, 11, 12 showed potent free radical scavenging activities. Also, for the cellular protective effects on HaCaT keratinocytes damaged by H2O2, the extract, EtOAc fraction and isolated compounds 10-12 indicated protective effects against oxidative stress. Upon the anti-bacterial tests using S. epidermidis and C. acnes the extract, n-Hex, EtOAc and n-BuOH fractions showed activities. In the anti-inflammatory tests using RAW264.7 cells, the n-Hex, EtOAc fraction, isolates 11 and 12 inhibited the production of NO without causing cell toxicity. Moreover, the compounds 11 and 12 exhibited the PGE2 and pro-inflammatory cytokines (TNF-α, IL-6) inhibition activity. Eight phytochemicals were isolated from the extract of Sorbus alnifolia branches; β-amyrin (1), ursolic acid (2), 2-oxopomolic acid (3), euscaphic acid (4), β-sitosterol (5), daucosterol (6), epi-catechin (7), prunasin (8). On the anti-oxidative tests, the extract, EtOAc and n-BuOH fractions showed potent free radical scavenging activities. Also, for the cellular protective effects on HaCaT keratinocytes damaged by H2O2, the EtOAc and n-BuOH fractions indicated protective effects against oxidative stress. Upon the anti-bacterial tests using S. epidermidis and C. acnes the extract, n-Hex, EtOAc and n-BuOH fractions and isolated compounds 3, 4 showed anti-bacterial activities. In the anti-inflammatory tests using RAW264.7 cells, the extract, n-Hex and EtOAc fractions, as well as isolates 3 and 4 inhibited the production of NO without causing cell toxicity. Moreover, the compound 3 exhibited PGE2 and pro-inflammatory cytokines (TNF-α, IL-6) inhibition activity and compound 4 only decreased production of (TNF-α, IL-6). Based on these results, it was suggested that the extract and isolated compounds from aerial parts of Epilobium angustifolium L., Chamaecyparis obusa seeds, Sorbus alnifolia leaves and branches could be potentially applicable as natural source for pharmaceutical and/or cosmetic ingredients.