• Bulletin of the Korean Chemical Society
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• 제34권10호
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• pp.3017-3021
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• 2013

Coumaroyl dipeptide amide, Coumaric acid-LG-$NH_2$, was prepared successfully using the solid-phase method, and its efficacy as a skin whitening agent was studied. Coumaric acid-LG-$NH_2$ was prepared with Rink-amide resin, and 96.354% of purity was obtained. Using MTT assay and LDH release assay, we found that it exhibited very low cytotoxicity. And, we found that Coumaric acid-LG-$NH_2$ inhibited tyrosinase activity dose-dependently and showed superior tyrosinase inhibitory activity to well-known whitening agent, arbutin. $IC_{50}$ value of Coumaric acid-LG-$NH_2$ was 182.4 ${\mu}M$, and $IC_{50}$ value of arbutin was 384.6 ${\mu}M$. Also, in measurement of melanin contents using B16F1 melanoma cell lines, Coumaric acid-LG-$NH_2$ reduced melanin production induced by ${\alpha}$-MSH statistically significant, and showed superior melanin inhibitory activity to p-coumaric acid or arbutin. In addition, Coumaric acid-LG-$NH_2$ reduced MC1R mRNA expression level. Thus, we concluded that MC1R pathway is the significant pathway of Coumaric acid-LG-$NH_2$, and Coumaric acid-LG-$NH_2$ has great potential to be used as novel whitening agents.

• 동의생리병리학회지
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• 제30권1호
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• pp.47-53
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• 2016

Kaempferia galanga L. is one of the plants in Zingiberaceae family. It is used by people in many regions of Asia and Africa for relieving toothache, abdominal pain, muscular swelling and rheumatism. Tyrosinase is a key enzyme for melanogenesis, and hyperpigmentation is associated with abnomal accumulation of melanin pigment. This study aimed to investigate the inhibition of melanogenesis by hexane extract of Kaempferia galanga L. (HKG) in B16F10 melanoma cells. Cell-free tyrosinase, melanin contents, intracellular tyrosinase activity and western blot analysis were performed to elucidate the effects on anti-melanogenesis. Cytotoxicity of the extracts was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and determined the concentration of 12.5, 25 μg/ml. HKG significantly inhibited to activities of intracellular tyrosinase and melanin synthesis in the absence or presence of α-melanocyte stimulating hormone (α-MSH) with dose-dependent manner. And HKG inhibited the expression of tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2), regardless of the presence or absence of α-MSH. HKG also down-regulated phosphorylation of p38 and JNK, and up-regulated phosphorylation of Akt. These effects were not related to its cytotoxicity action. These results indicate that HKG has the potential to be a useful therapeutic agent for treating hyperpigmentation disorders and as a beneficial additive in whitening agents in cosmetics industry.

• 한국응용과학기술학회지
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• 제39권5호
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• pp.710-719
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• 2022

본 연구는 Passiflora caerulea 추출물의 항산화 활성 및 멜라닌 생합성 억제 효과 측정을 통해 화장품 소재로서의 활용 가능성을 확인하고자 하였다. Passiflora caerulea 추출물은 70% 에탄올로 추출하였고, TEAC assay를 이용한 ABTS radical 소거능을 측정하였다. 또한 Neutral red assay를 이용한 세포 생존율, DCF-DA를 통한 세포 내 ROS 생성 억제, 멜라닌 생합성 억제 효과를 측정하였다. 연구결과, Passiflora caerulea 추출물의 ABTS radical 소거능은 농도 의존적 소거활성이 확인되었으며, positive control(양성대조군)로 이용한 trolox와 0.1 mg/mL 농도에서는 유사한 radical 소거 활성을 확인하였다. CFDA를 통한 세포 내 활성산소종(ROS) 생성은 농도 의존적으로 억제되었고, B16F10 melanoma 세포에 대한 세포 독성은 나타나지 않았다. 또한 𝛼-MSH로 유도된 멜라닌 생합성 억제 효과를 확인하였다. 따라서 본 연구를 통해 Passiflora caerulea 추출물은 화장품 소재로서의 활용 가능성이 있는 것으로 판단된다.

• 생약학회지
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• 제53권2호
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• pp.70-78
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• 2022

The objective of this study was to assess the effect of preventing pigmentation caused by external stimuli, promoting skin regeneration and whitening of the skin composition containing complex extract of Dendrobii Caulis and Centella asiatica (CE). We evaluated cell viability, tyrosinase and melanin inhibition activity, skin irritation test, and skin moisturizing and regenerative effects using CE. As a result of the tyrosinase inhibitory activity, 100 ㎍/mL CE (35.31%) showed higher value than kojic acid (21.32%). The results of melanin synthesis inhibition in B16F10 melanoma cells after treatment with α-melanocyte stimulating hormone showed a similar level of activity to that of arbutin, indicating an excellent whitening effect. In clinical test of the skin composition containing CE, we confirmed that CE is non-irritated in human skin primary irritation test as well as have a high skin moisturizing and regenerative effect. From these results, we suggested the CE not only prevents skin damage and pigmentation caused by external stimuli but has remarkable skin brightening activity and skin regeneration effect.

• 대한본초학회지
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• 제20권4호
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• pp.17-26
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• 2005

Objectives : Antioxidant, Anticancer and Antibacterial Activities of Naesohwangryntang and its composition oriental medicines. Methods : We were experimented anti-oxidation effect and growth inhibition ability on cancer cells and antibacterial activity on various kinds of bacteria of skin. Results : The results were obtained as follows : Electron donating ability(EDA) of water extract Naesohwangryntang and ethanol extract Naesohwangryntang was 60% and 70% at 1000 ppm concentration. In the test of SOD-like activity, ethanol extract showed more activity with 27.4% in 700 ppm, while water extract was low in 19.6%. Clear zones formed by sample against the human skin-resident microflora indicated that anti-microbial activity of ethanol extract Naesohwangryntang was higher than that of water extract Naesohwangryntang. The growth inhibition rates of each sample on lung-cancer(A549), at 1000 ppm cancer cell was over 40%. The growth inhibition rate of the each sample melanoma-cancer(B16F10, G361), at 1000 ppm was over 80%. Conclusions : The results indicated that, ethanol extract which is superior in its anti-oxidation and antibacterial effect is useful to be applied in cosmetic industry.

• ALGAE
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• 제28권1호
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• pp.111-119
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• 2013

Marine microalgae are a promising source of organisms that can be cultured and targeted to isolate the broad spectrum of functional metabolites. In this study, two species of cyanobacteria, Chlorella ovalis Butcher and Nannchloropsis oculata Droop, one species of bacillariophyta, Phaeoductylum tricornutum Bohlin, and one species of Dinophyceae, Amphidinium carterae (Hulburt) were cultured and biomasses used to evaluate the proximate comical compositions. Among the determined proximate chemical compositions of the cultured marine microalgae, the highest content of crude proteins and lipids were exhibited in P. tricornutum and A. carterae, respectively. Solvent-solvent partition chromatography was subjected to fractionate each of the cultured species and separated n-hexane, chloroform, ethyl acetate, and aqueous fractions. Nitric oxide production inhibitory level (%) and cytotoxicity effect on lipo-polysaccharide-induced RAW 264.7 macrophages were performed to determine the anti-inflammatory activity. N. oculata hexane and chloroform fractions showed significantly the strongest anti-inflammatory activity at $6.25{\mu}g\;mL^{-1}$ concentration. The cancer cell growth inhibition (%) was determined on three different cell lines including HL-60 (a human promyelocytic leukemia cell line), A549 (a human lung carcinoma cell line), and B16F10 (a mouse melanoma cell line), respectively. Among the extracts, C. ovalis ethyl acetate and A. carterae chloroform fractions suppressed the growth of HL-60 cells significantly at 25 and $50{\mu}g\;mL^{-1}$ concentrations. Thus, the cultured marine microalgae solvent extracts may have potentiality to isolate pharmacologically active metabolites further using advance chromatographic steps. Hence, the cultured marine microalgae can be described as a good candidate for the future therapeutic uses.

• IMMUNE NETWORK
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• 제22권5호
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• pp.42.1-42.20
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• 2022

Vaccination with tumor peptide epitopes associated with MHC class I molecules is an attractive approach directed at inducing tumor-specific CTLs. However, challenges remain in improving the therapeutic efficacy of peptide epitope vaccines, including the low immunogenicity of peptide epitopes and insufficient stimulation of innate immune components in vivo. To overcome this, we aimed to develop and test an innovative strategy that elicits potent CTL responses against tumor epitopes. The essential feature of this strategy is vaccination using tumor epitope-loaded nanoparticles (NPs) in combination with polyinosinic-polycytidylic acid (poly-IC) and anti-PD1 mAb. Carboxylated NPs were prepared using poly(lactic-co-glycolic acid) and poly(ethylene/maleic anhydride), covalently conjugated with anti-H-2K^b mAbs, and then attached to H-2K^b molecules isolated from the tumor mass (H-2^b). Native peptides associated with the H-2K^b molecules of H-2K^b-attached NPs were exchanged with tumor peptide epitopes. Tumor peptide epitope-loaded NPs efficiently induced tumor-specific CTLs when used to immunize tumor-bearing mice as well as normal mice. This activity of the NPs significantly was increased when co-administered with poly-IC. Accordingly, the NPs exerted significant anti-tumor effects in mice implanted with EG7-OVA thymoma or B16-F10 melanoma, and the anti-tumor activity of the NPs was significantly increased when applied in combination with poly-IC. The most potent anti-tumor activity was observed when the NPs were co-administered with both poly-IC and anti-PD1 mAb. Immunization with tumor epitope-loaded NPs in combination with poly-IC and anti-PD1 mAb in tumor-bearing mice can be a powerful means to induce tumor-specific CTLs with therapeutic anti-tumor activity.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.98-98
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• 2003

${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$＋＋/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

• 한국미생물·생명공학회지
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• 제40권4호
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• pp.371-379
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• 2012

본 연구에서는 한약재로 사용되고 있는 산초 과피 추출물의 기능성 소재로서의 활용 가능성을 알아보기 위해 산초 추출물 및 분획물이 보유한 항산화, 미백 및 항염증 활성을 in vitro assay system 및 cell culture model system을 이용하여 분석하였다. 그 결과 산초 MeOH, EtOH, 열수 추출물 모두 강한 DPPH radical 소거 활성 및 tyrosinase 효소 활성 저해를 통한 DOPA 산화 억제능을 보였다. B16F10 mouse melanoma cell을 이용하여 ${\alpha}$-MSH에 의해 유도된 세포 내 melanin 생성에 미치는 산초 추출물의 영향을 알아본 결과 농도의존적인 melanin 생성 억제능을 보였고 세포의 tyrosinase 활성 저해능 또한 이와 일치하는 결과를 보여 산초 추출물에 의한 melanin 생성 억제능이 tyrosinase 효소 활성 억제 및 melanogenesis 관련 단백질 발현 저해에서 기인한 것으로 판단된다. 또한 RAW 264.7 murine macrophage cell을 이용하여 산초 추출물의 항염증 활성을 분석한 결과 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 단백질 발현 분석을 통해 확인하였다. 산초 추출물에 존재하는 활성물질을 규명하기 위해 용매 분획을 실시한 후 각 분획물의 미백 및 항염증 활성을 비교 분석한 결과 물 층을 제외한 모든 용매 분획이 활성을 보유함을 확인하였다. 이러한 결과를 통해 산초 추출물이 높은 항산화능과 미백활성, 그리고 항염증 활성을 보유함을 확인할 수 있었으며, 이러한 결과는 미백활성을 비롯한 생리활성 보유 천연물 소재 탐구의 기초자료로 유용하게 쓰일 것으로 사료된다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.21-22
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• 2019

Melanin is a mixture of pigmented biopolymers synthesized by epidermal melanocytes that determine the skin, eye, and hair colors. Melanocytes produce two different kinds of melanin, eumelanin (dark brown/black insoluble pigments found in dark skin and dark hair and pheomelanin (lighter red/yellow). The biological role of melanin is to prevent skin damage by ultraviolet (UV) radiation. However, the overproduction or deficiency of melanin synthesis could lead to serious dermatological problems, which include melasma, melanoderma, lentigo, and vitiligo. Therefore, regulating melanin production is important to prevent the pigmentation disorders. Myanmar has a rich in natural resources. However, the chemical constituents of these natural resources in Myanmar have not been fully investigated. In the effort to search for compounds with anti-melanin deposition activity from Myanmar natural resources, five plants were collected in Myanmar. Extracts of these collected five plants were tested for anti-melanin deposition activity against a mouse melanoma cell line (B16-F10) induced with ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) and 3-isobutyl-1-methylxanthine (IBMX), and their anti-melanin deposition activities were compared with the positive control, arbutin. Among the tested extracts, the CHCl3 extracts of the Premna serratifolia (syn: P. integrifolia) wood showed anti-melanin deposition activities with IC50 values of $81.3{\mu}g/mL$. Hence, this study aims to identify secondary metabolites with anti-melanin deposition activity from P. serratifolia wood of Myanmar. P. serratifolia belongs to the Verbenaceae family and is widely distributed in near western sea coast from South Asia to South East Asia, which include India, Malaysia, Vietnam, Cambodia, and Sri Lanka. People in Tanintharyi region located in the southern part of Myanmar utilize the P. serratifolia, Sperethusa crenulata, Naringi crenulata, and Limonia acidissima as Thanaka, traditional cosmetics in Myanmar. Thanaka is applied in the form of paste onto skins to make it smooth and clear, as well as to prevent wrinkles, skin aging, excessive facial oil, pimples, blackheads, and whiteheads. However, the chemical constituents responsible for their cosmetic properties are yet to be identified. Moreover, the chemical constituents of P. serratifolia was almost uncharacterized. Investigation of the P. serratifolia chemical constituents is thus an attractive endeavor to discover new anti-melanin deposition active compounds. The investigation of the chemical constituents of the active CHCl3 extract of P. serratifolia led to isolation of four new lignoids, premnan A (1), premnan B (2), taungtangyiol C (3), and 7,9-dihydroxydolichanthin B (4), together with premnan C (5) (assumed to be an artifact), one natural newlignoid,(3R,4S)-4-(1,3-benzodioxol-5-ylcarbonyl)-3-[(R)-1-(1,3-benzo dioxol-5-yl)-1-hydroxy methyl]tetrahydro-2-furanone (6), and five known compounds (7-11)1,2). The structures of all isolated compounds were determined on the basis of their spectroscopic data and by comparison with the reported literatures. The absolute configurations of 1-3 and 5 were also determined by optical rotation and circular dichroism (CD) data analyses1). The anti-melanin deposition activities of all the isolated compounds were evaluated against B16-F10 cell line. 7,9-Dihydroxydolichanthin B (4) and ($2{\alpha},3{\alpha}$)-olean-12-en-28-oic acid (11) showed strong anti-melanin deposition activities with IC50 values of 18.4 and $11.2{\mu}M$, respectively, without cytotoxicity2). On the other hand, compounds 1-3, 5, and 7 showed melanogenesis enhancing activities1). To better understand their anti-melanin deposition mechanism, the effects of 4 and 11 on tyrosinase activities were investigated. The assay indicated that compounds 4 and 11 did not inhibit tyrosinase. Furthermore, we also examined the mRNA expression of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2). Compounds 4 and 11 down-regulated the expression of Tyr and Mitf mRNAs, respectively. Although the P. serratifolia wood has been used as traditional cosmetics in Myanmar for centuries, there are no scientific evidences to support its effectiveness as cosmetics. Investigation of the anti-melanin deposition activity of the chemical constituents of P. serratifolia thus provided insight into the effectiveness of the P. serratifolia wood as a cosmetic agent.