WO2018066704A1 - Melanin inhibitor, antibacterial agent, antiallergic agent, accelerator for collagen production, and moisturizer - Google Patents
Melanin inhibitor, antibacterial agent, antiallergic agent, accelerator for collagen production, and moisturizer Download PDFInfo
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- WO2018066704A1 WO2018066704A1 PCT/JP2017/036530 JP2017036530W WO2018066704A1 WO 2018066704 A1 WO2018066704 A1 WO 2018066704A1 JP 2017036530 W JP2017036530 W JP 2017036530W WO 2018066704 A1 WO2018066704 A1 WO 2018066704A1
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- A—HUMAN NECESSITIES
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/63—Steroids; Derivatives thereof
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/04—Antipruritics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
Definitions
- the present invention relates to a melanin inhibitor, an antibacterial agent, an antiallergic agent, a collagen production promoting agent, and a moisturizing agent.
- Biwa is a subtropical evergreen tree belonging to the Rosaceae family. Biwa leaves have been used in traditional Chinese medicine. In Japan, loquat leaves are used as Biwa tea either alone or mixed with tea leaves. Patent Document 1 shows that loquat leaf extract can be used for the treatment and prevention of osteopenic diseases.
- an object of the present invention is to provide a safe compound of natural origin that can be used in the fields of skin care such as whitening, acne prevention, anti-allergy, wrinkle prevention, and skin moisturizing.
- the present invention is characterized in that the melanin inhibitor contains a triterpenoid extracted from loquat leaves as an active ingredient.
- the present invention is characterized in that the antibacterial agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
- the present invention is characterized in that the antiallergic agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
- the present invention is characterized in that the collagen production promoting agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
- the present invention is characterized in that the moisturizing agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
- a safe compound of natural origin that can be used in the fields of skin care such as whitening, acne prevention, anti-allergy, wrinkle prevention, and skin moisturizing.
- Embodiments of a melanin inhibitor, an antibacterial agent, an antiallergic agent, a collagen production promoter and a moisturizer according to the present invention will be described. This embodiment is merely an example, and the present invention is not limited to this.
- Loquat, Eriobotrya It is a flowering plant of the Rosaceae known as japonica.
- Biwa leaves are known as traditional Chinese medicine. In Japan, Biwa leaves are also drunk as Biwa tea. Biwa has been confirmed to suppress obesity and tumors by numerous chemical pharmacological studies. Therefore, we investigated the utility of loquat leaves for skin care.
- Triterpenoids Eighteen types of triterpenoids that are loquat leaf extracts were evaluated for their effects on skin care. These 18 types of triterpenoids are (1) ursolic acid, (2) oleanolic acid, (3) uvaol, (4) betulinic acid, and (5) muslin.
- the numbers enclosed in parentheses are reference compound (triterpenoid) numbers.
- maslinic acid is indicated.
- mice B16 melanoma cells were obtained from the RIKEN BioResource Center cell bank. Cell survival was maintained in Eagle's minimum essential medium supplemented with 0.09 mg / ml theophylline and 10% (v / v) fetal calf serum. Prior to sample processing, B16 melanoma cells were incubated for 24 hours in medium at a cell concentration of 1 ⁇ 10 5 (cells / well).
- DMSO dimethyl sulfoxide
- B16 melanoma cells were further incubated for 48 hours, after which the medium was again replaced with fresh medium containing each sample. After another 24 hours, melanin content and cell viability were measured using a microplate reader at 405 nm and 570 nm, respectively. Cell viability was measured using the MTT (3- (4,5-dimethylthiazol-2-yl) -2,5 diphenyl bromide) method.
- the inhibitory effect on melanin synthesis and the cytotoxic effect on cell viability of 16 compounds extracted from loquat leaves were tested.
- the value of the cytotoxic concentration CC 50 causing death in 50% of viable cells and the concentration IC 50 causing 50% inhibition of melanin synthesis in B16 melanoma cells were calculated from the corresponding dose response curves.
- the values for CC 50 and IC 50 are shown in Table 1.
- CC 50 / IC 50 The relative efficacy of compounds with respect to the inhibition of melanogenesis against the induction of cell death, ie the value of CC 50 / IC 50 is defined as the selectivity index (SI) and is shown together in Table 1. It was. Situations where melanin synthesis is inhibited include situations where B16 melanoma cells die, in addition to situations where melanin production is simply inhibited. CC 50 indicates that the B16 melanoma cells die, that is, the magnitude of the cytotoxic effect. IC 50 shows the magnitude of the inhibitory effect on melanin synthesis including cytotoxic effects.
- SI selectivity index
- melanin-suppressing effect selectively inhibits melanin production, that is, it does not cause cytotoxicity unless it is at a higher concentration, and inhibits melanin production even at a lower concentration (S. I.> 1).
- some of the triterpenoids extracted from loquat leaves that is, compounds 1, 2, 4, 5, 6, 7, 8, and 11 inhibit melanin production. That is, among triterpenoids extracted from loquat leaves, a compound selected from the group consisting of ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolic acid, maslinic acid, corosolic acid, tolmenic acid, or a salt thereof Solvates or hydrates can be used as safe melanin inhibitors of natural origin.
- triterpenoids extracted from loquat leaves especially ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolic acid, maslinic acid, corosolic acid, and tolmenic acid, are used for skin care with a function of inhibiting melanin production, that is, whitening. It can be used as an agent.
- [antibacterial] Propionibacterium acnes was used as a test bacterium for evaluating the antibacterial properties of 18 types of triterpenoids extracted from loquat leaves. A single colony of the test bacterium was collected, added to the medium, and cultured at 37 ° C. for 30 hours. This was then added to the bacterial suspension and adjusted to a bacterial concentration of 10 5 CFU / ml.
- each compound was dissolved in DMSO. Sorbic acid was used as a control sample. Furthermore, each compound dissolved in DMSO was incubated at 37 ° C. for 30 hours.
- MIC Minimum inhibitory concentration
- MMC minimum bactericidal concentration
- Triterpenoids exhibit low hydrophilicity with log P values ranging from 5 to 8. In general, antibacterial activity tends to be low when the hydrophilicity is low, but compounds 1, 2, 6, 7, 8, 10, 11, 12, 14, 16, 17, and 18 have high antibacterial activity. Shows activity.
- some of the triterpenoids extracted from loquat leaves that is, compounds 1, 2, 6, 7, 8, 10, 11, 12, 14, 16, 17, 18 are propionibacterium acne. It can be seen that it has antibacterial activity against bacteria. That is, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, tolmenic acid, oiscafic acid, 6 ⁇ , 19-dihydroxyurs-12-en-3
- the salt, solvate or hydrate can be used as a safe antibacterial agent of natural origin.
- Propionibacterium acne is an anaerobic gram-positive bacterium mainly located in the pilosebaceous unit and plays an important role in the development of acne. Therefore, some of the triterpenoids extracted from leaf leaves are effective in suppressing the development of acne.
- ursolic acid oleanolic acid
- methyl corosolic acid maslinic acid
- corosolic acid pomol acid
- tolmenic acid oiscafic acid
- 6 ⁇ 19-dihydroxyurs-12-en-3 -Oxo-28-Oic acid
- Hyptadienic acid 3-O-trans-p-coumaroyl tormic acid
- 3-O-cis-p-coumaroyl tormentic acid are antibacterial agents that suppress the development of acne It can be used as a skin care agent having the function of
- RBL-2H3 IgE-stimulated rat basophilic leukemia
- RBL-2H3 cells were maintained in EMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 37 ° C. in a humidified air incubator with a CO 2 concentration of 5%. The cell concentration was 2 ⁇ 10 5 cells / well. After 24 hours of incubation, the medium was supplemented with 1 ml of 0.5 ⁇ g / anti-DNP (dinitrophenol) mouse immunoglobulin E (IgE. After further incubation of the culture for 24 hours, the medium was added with 100 ⁇ l of thyroid buffer. Replaced with liquid.
- IgE dinitrophenol mouse immunoglobulin E
- the sample was dissolved in 1 ⁇ l DMSO at a concentration of less than 0.5%, and the cell plate was further incubated for 30 minutes.
- the sample containing thyroid buffer was then replaced with 100 ⁇ l of thyroid buffer containing 50 ⁇ g / ml DNP-BSA. After 1 hour incubation, 50 ⁇ l of the supernatant was collected from each well and each was mixed with 50 ⁇ l of 2M substrate solution (p-nitrophenyl-N-acetyl- ⁇ -glucosaminide).
- the mixture was incubated for 3 hours on a shaker at room temperature. Finally, 100 ⁇ l was added using 100 mM sodium bicarbonate as a stop solution. Thereafter, the absorbance was read at 405 nm. Cell viability was measured by the same MTT test as described above. The compound concentration was 25 ⁇ g / ml. Quercetin was used as a control sample.
- concentration IC 50 values that cause hexosaminidase release inhibition are measured and are 39.5 ⁇ M, 22.86 ⁇ M, 25.6 ⁇ M, and 15.8 ⁇ M, respectively. I found out.
- Compounds 1, 7, 9, and 12 are those that cause inflammation caused by 12-O-tetradecanoylphorbol-13-acetate (TPA) in mice and TPA-induced It can be seen that it has significant inhibitory activity against the activation of Epstein-Barr virus early antigen (EBV-EA).
- TPA 12-O-tetradecanoylphorbol-13-acetate
- triterpenoids extracted from loquat leaves that is, compounds 1, 7, 9, and 12 have anti-allergic properties. That is, among triterpenoids extracted from loquat leaves, a compound selected from the group consisting of ursolic acid, maslinic acid, 3-epicorosolic acid, oiscafic acid, or a salt, solvate or hydrate thereof, It can be used as a safe antiallergic agent of natural origin.
- DMEM Dulbecco's Modified Eagle Medium
- the medium was replaced with a 99 microliter mixture of DMEM supplemented with 1 ⁇ l of sample solution and 0.5% FBS.
- the DMSO concentration was 0.5% in the medium.
- the amount of type I collagen produced in the medium was measured using an ELISA kit manufactured by ACEL. Collagen production was calculated using a standard curve created by measuring with the same ELISA kit. The remaining cells were used for MTT evaluation. 100 ⁇ M ascorbic acid was used as a control sample.
- CP collagen production activity
- CV cell viability
- some triterpenoids significantly increase collagen production of dermal fibroblasts per cell.
- 10 types of triterpenoids extracted from loquat leaves 10 types of compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, and 14 produced type I collagen at a concentration of 10 ⁇ g / ml.
- Compounds 6, 8 show particularly strong stimulating activity of 3.4 and 3.6, which is higher than the control sample. Other compounds show a moderate promoting effect.
- some of the triterpenoids extracted from loquat leaves ie, compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17 , 18 have collagen generation-promoting properties.
- ursolic acid, oleanolic acid, ubaol, betulinic acid methyl masphosphate, methyl corosolic acid, maslinic acid, corosolic acid, 3-epicorosolic acid, pomolic acid, tolmenic acid, Oiskafic acid, 3-oxourus-12-en-28-oic acid, 6 ⁇ , 19-dihydroxyurs-12-en-3-oxo-28-oic acid, 2 ⁇ , 19 ⁇ -dihydroxy-3-oxo-12-ursen-
- the dermal-epidermal junction is flattened and mechanical tension and elasticity are reduced due to a marked decrease in the extracellular matrix of skin such as collagen, elastin and hyaluronic acid.
- Some of the triterpenoids extracted from loquat leaves are effective in producing collagen, and as a result, the generation of wrinkles in the skin can be suppressed.
- ursolic acid among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl masphosphate, methyl corosolic acid, maslinic acid, corosolic acid, 3-epicorosolic acid, pomolic acid, tolmenic acid, Oiskafic acid, 3-oxourus-12-en-28-oic acid, 6 ⁇ , 19-dihydroxyurs-12-en-3-oxo-28-oic acid, 2 ⁇ , 19 ⁇ -dihydroxy-3-oxo-12-ursen- 28-Oic acid, 3-O-trans-p-coumaroyl tormentic acid, 3-O-cis-p-coumaroyl tormentic acid, stimulates collagen production and suppresses skin aging It can be used as a skin care agent having the function of an agent.
- DMEM Dulbecco's Modified Eagle Medium
- hyaluronic acid in the medium was measured using a QnE hyaluronic acid ELISA kit manufactured by Bio Trading Partners. Hyaluronic acid production was calculated from a quantitative standard curve. Cell viability was measured by the MTT method. Ascorbic acid was used as a control sample. For each compound, HA / CV values at three compound concentrations of 1, 5 and 10 ⁇ g / ml were determined.
- the HA / CV value is a hyaluronic acid production promotion rate, and is an average value of hyaluronic acid concentration (%) / average value of cell viability (%).
- Hyaluronic acid concentration (HA) and cell viability (CV) values were determined as a percentage of the control sample.
- some triterpenoids significantly increase hyaluronic acid production of dermal fibroblasts per cell.
- 10 types of triterpenoids extracted from loquat leaves 10 types of compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, 18 have a concentration of 10 ⁇ g / ml in human skin. It exerted a strong stimulating effect of hyaluronic acid production in fibroblasts.
- Compounds 8, 10 showed stimulating activity values of 5.9 and 5.8 over the control sample.
- Compound 6 also shows a high stimulating activity value.
- Compound 1 has a stimulating activity value of 4.7 exceeding the comparative example.
- some of the triterpenoids extracted from loquat leaves that is, compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, 18 have hyaluronic acid production promoting properties. . That is, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, 3-oxours-12-en-28-oic acid, A compound selected from the group consisting of 3-O-cis-p-coumaroyl tormentic acid, or a salt, solvate or hydrate thereof, should be used as a safe hyaluronic acid production accelerator of natural origin. Can do.
- the youthful skin contains a lot of moisture and maintains swell pressure, elasticity and flexibility.
- Hyaluronic acid has a high ability to retain moisture and greatly affects the amount of moisture in the skin.
- Some of the triterpenoids extracted from loquat leaves are effective in producing collagen and, as a result, are effective in maintaining youthful skin.
- ursolic acid oleanolic acid, ubaol, betulinic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, 3-oxours-12-en-28-oic acid, 3-O-cis-p-coumaroyl tormentic acid can be used as a skin care agent having a function as a moisturizing agent for increasing the moisture content of the skin.
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Abstract
[Problem] To provide a safe compound having originated from the natural world, the compound being usable in the skin care fields, for example, skin whitening, acne prevention, anti-inflammation, wrinkle prevention, and skin moisturization.
[Solution] A melanin inhibitor which contains one or more triterpenoids extracted from loquat leaves, in particular, one or more compounds selected from the group consisting of ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolate, maslinic acid, corosolic acid, and tormentic acid. These compounds may be salts, solvates, or hydrates thereof.
Description
本発明は、メラニン抑制剤、抗菌剤、抗アレルギー剤、コラーゲン生成促成剤および保湿剤に関する。
The present invention relates to a melanin inhibitor, an antibacterial agent, an antiallergic agent, a collagen production promoting agent, and a moisturizing agent.
びわは、バラ科に属する亜熱帯常緑樹である。びわの葉は、伝統的な中国医学で用いられてきた。日本では、ビワの葉は、単体で、あるいは茶葉と混ぜて、びわ茶として用いられている。特許文献1では、びわの葉の抽出物が骨減少性疾患の治療および予防のために用いることができることが示されている。
Biwa is a subtropical evergreen tree belonging to the Rosaceae family. Biwa leaves have been used in traditional Chinese medicine. In Japan, loquat leaves are used as Biwa tea either alone or mixed with tea leaves. Patent Document 1 shows that loquat leaf extract can be used for the treatment and prevention of osteopenic diseases.
美白、にきび予防、抗炎症、しわの発生予防、肌の保湿などのスキンケアの分野では、様々な化粧料が開発されている。
Various cosmetics have been developed in the fields of skin care such as whitening, acne prevention, anti-inflammation, wrinkle prevention, and skin moisturizing.
そこで、本発明は、美白、にきび予防、抗アレルギー、しわの発生予防、肌の保湿などのスキンケアの分野において用いることができる自然界起源の安全な化合物を提供することを目的とする。
Therefore, an object of the present invention is to provide a safe compound of natural origin that can be used in the fields of skin care such as whitening, acne prevention, anti-allergy, wrinkle prevention, and skin moisturizing.
上述の目的を達成するため、本発明は、メラニン抑制剤において、びわの葉から抽出されるトリテルペノイドを有効成分として含有することを特徴とする。
In order to achieve the above object, the present invention is characterized in that the melanin inhibitor contains a triterpenoid extracted from loquat leaves as an active ingredient.
また、本発明は、抗菌剤において、びわの葉から抽出されるトリテルペノイドを有効成分として含有することを特徴とする。
Further, the present invention is characterized in that the antibacterial agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
また、本発明は、抗アレルギー剤において、びわの葉から抽出されるトリテルペノイドを有効成分として含有することを特徴とする。
Further, the present invention is characterized in that the antiallergic agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
また、本発明は、コラーゲン生成促成剤において、びわの葉から抽出されるトリテルペノイドを有効成分として含有することを特徴とする。
Further, the present invention is characterized in that the collagen production promoting agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
また、本発明は、保湿剤において、びわの葉から抽出されるトリテルペノイドを有効成分として含有することを特徴とする。
Further, the present invention is characterized in that the moisturizing agent contains a triterpenoid extracted from loquat leaves as an active ingredient.
本発明によれば、美白、にきび予防、抗アレルギー、しわの発生予防、肌の保湿などのスキンケアの分野において用いることができる自然界起源の安全な化合物を提供することができる。
According to the present invention, it is possible to provide a safe compound of natural origin that can be used in the fields of skin care such as whitening, acne prevention, anti-allergy, wrinkle prevention, and skin moisturizing.
本発明に係るメラニン抑制剤、抗菌剤、抗アレルギー剤、コラーゲン生成促成剤、保湿剤の一実施の形態を、説明する。なお、この実施の形態は単なる例示であり、本発明はこれに限定されない。
Embodiments of a melanin inhibitor, an antibacterial agent, an antiallergic agent, a collagen production promoter and a moisturizer according to the present invention will be described. This embodiment is merely an example, and the present invention is not limited to this.
びわは、loquat, Eriobotrya
japonicaとして知られるバラ科の顕花植物である。びわの葉は、伝統的漢方薬として知られている。また、日本国内でも、びわの葉は、びわ茶として飲用されている。びわは、数多くの化学薬理学研究によって、肥満抑制、腫瘍抑制などが確かめられている。そこで、びわの葉のスキンケアに対する効用を調べた。 Loquat, Eriobotrya
It is a flowering plant of the Rosaceae known as japonica. Biwa leaves are known as traditional Chinese medicine. In Japan, Biwa leaves are also drunk as Biwa tea. Biwa has been confirmed to suppress obesity and tumors by numerous chemical pharmacological studies. Therefore, we investigated the utility of loquat leaves for skin care.
japonicaとして知られるバラ科の顕花植物である。びわの葉は、伝統的漢方薬として知られている。また、日本国内でも、びわの葉は、びわ茶として飲用されている。びわは、数多くの化学薬理学研究によって、肥満抑制、腫瘍抑制などが確かめられている。そこで、びわの葉のスキンケアに対する効用を調べた。 Loquat, Eriobotrya
It is a flowering plant of the Rosaceae known as japonica. Biwa leaves are known as traditional Chinese medicine. In Japan, Biwa leaves are also drunk as Biwa tea. Biwa has been confirmed to suppress obesity and tumors by numerous chemical pharmacological studies. Therefore, we investigated the utility of loquat leaves for skin care.
[評価対象]
びわの葉の抽出物である18種類のトリテルペノイドについてスキンケアに対する効用に関する評価を行った。この18種類のトリテルペノイドとは、(1)ウルソル酸(ursolic acid)、(2)オレアノール酸(oleanolic acid)、(3)ウバオール(uvaol)、(4)ベツリン酸(betulinic acid)、(5)マスリン酸メチル(methyl maslinate)、(6)コロソリン酸メチル(methyl corosolate)、(7)マスリン酸(maslinc acid)、(8)コロソリン酸(corosolic acid)、(9)3-エピコロソリン酸(3-epicorosolic acid)、(10)ポモル酸(pomolic acid)、(11)トルメン酸(tormentic acid)、(12)オイスカフ酸(euscaphic acid)、(13)3-オキソウルス-12-エン-28-オイック酸(3-oxours-12-en-28-oic
acid)、(14)6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸(6β,19-dihydroxyurs-12-en-3-oxo-28-oic acid)、(15)2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸(2α,19α-dihydroxy-3-oxo-12-ursen-28-oic
acid)、(16)ヒプタジエニック酸(hyptadienic acid)、(17)3-O-トランス-p-クマロイルトルメンチック酸(3-O-trans-p-coumaroyltormentic acid)、(18)3-O-シス-p-クマロイルトルメンチック酸(3-O-cis-p-coumaroyltormentic
acid)、である。 [Target]
Eighteen types of triterpenoids that are loquat leaf extracts were evaluated for their effects on skin care. These 18 types of triterpenoids are (1) ursolic acid, (2) oleanolic acid, (3) uvaol, (4) betulinic acid, and (5) muslin. Methyl maslinate, (6) methyl corosolate, (7) maslinc acid, (8) corosolic acid, (9) 3-epicorosolic acid acid), (10) pomolic acid, (11) tormentic acid, (12) euscaphic acid, (13) 3-oxours-12-en-28-oic acid (3) -oxours-12-en-28-oic
acid), (14) 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid, (15) 2α , 19α-Dihydroxy-3-oxo-12-ursen-28-oic acid (2α, 19α-dihydroxy-3-oxo-12-ursen-28-oic
acid), (16) hyptadienic acid, (17) 3-O-trans-p-coumaroyltormentic acid, (18) 3-O- Cis-p-coumaroyltormentic acid (3-O-cis-p-coumaroyltormentic
acid).
びわの葉の抽出物である18種類のトリテルペノイドについてスキンケアに対する効用に関する評価を行った。この18種類のトリテルペノイドとは、(1)ウルソル酸(ursolic acid)、(2)オレアノール酸(oleanolic acid)、(3)ウバオール(uvaol)、(4)ベツリン酸(betulinic acid)、(5)マスリン酸メチル(methyl maslinate)、(6)コロソリン酸メチル(methyl corosolate)、(7)マスリン酸(maslinc acid)、(8)コロソリン酸(corosolic acid)、(9)3-エピコロソリン酸(3-epicorosolic acid)、(10)ポモル酸(pomolic acid)、(11)トルメン酸(tormentic acid)、(12)オイスカフ酸(euscaphic acid)、(13)3-オキソウルス-12-エン-28-オイック酸(3-oxours-12-en-28-oic
acid)、(14)6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸(6β,19-dihydroxyurs-12-en-3-oxo-28-oic acid)、(15)2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸(2α,19α-dihydroxy-3-oxo-12-ursen-28-oic
acid)、(16)ヒプタジエニック酸(hyptadienic acid)、(17)3-O-トランス-p-クマロイルトルメンチック酸(3-O-trans-p-coumaroyltormentic acid)、(18)3-O-シス-p-クマロイルトルメンチック酸(3-O-cis-p-coumaroyltormentic
acid)、である。 [Target]
Eighteen types of triterpenoids that are loquat leaf extracts were evaluated for their effects on skin care. These 18 types of triterpenoids are (1) ursolic acid, (2) oleanolic acid, (3) uvaol, (4) betulinic acid, and (5) muslin. Methyl maslinate, (6) methyl corosolate, (7) maslinc acid, (8) corosolic acid, (9) 3-epicorosolic acid acid), (10) pomolic acid, (11) tormentic acid, (12) euscaphic acid, (13) 3-oxours-12-en-28-oic acid (3) -oxours-12-en-28-oic
acid), (14) 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid, (15) 2α , 19α-Dihydroxy-3-oxo-12-ursen-28-oic acid (2α, 19α-dihydroxy-3-oxo-12-ursen-28-oic
acid), (16) hyptadienic acid, (17) 3-O-trans-p-coumaroyltormentic acid, (18) 3-O- Cis-p-coumaroyltormentic acid (3-O-cis-p-coumaroyltormentic
acid).
ここで、括弧で閉じた数字は、参照用の化合物(トリテルペノイド)の番号である。たとえば、以下で化合物7と表現した場合、マスリン酸を示す。
Here, the numbers enclosed in parentheses are reference compound (triterpenoid) numbers. For example, when expressed as compound 7 below, maslinic acid is indicated.
それぞれの化合物の構造は以下の通りである。
The structure of each compound is as follows.
(14)6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸(6β,19-dihydroxyurs-12-en-3-oxo-28-oic
acid)
(14) 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid (6β, 19-dihydroxyurs-12-en-3-oxo-28-oic
acid)
acid)
acid)
(15)2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸(2α,19α-dihydroxy-3-oxo-12-ursen-28-oic
acid)
(15) 2α, 19α-dihydroxy-3-oxo-12-ursen-28-oic acid (2α, 19α-dihydroxy-3-oxo-12-ursen-28-oic
acid)
acid)
acid)
(17)3-O-トランス-p-クマロイルトルメンチック酸(3-O-trans-p-coumaroyltormentic acid)
(17) 3-O-trans-p-coumaroyltormentic acid
(18)3-O-シス-p-クマロイルトルメンチック酸(3-O-cis-p-coumaroyltormentic acid)
(18) 3-O-cis-p-coumaroyltormentic acid
[メラミン生成抑制]
びわの葉から抽出された18種類のトリテルペノイドのメラニン抑制効果を評価するため、マウスのB16メラノーマ細胞を用いた試験を行った。マウスのB16メラノーマ細胞は、理研バイオリソースセンター細胞バンクから入手した。濃度0.09mg/mlのテオフィリンおよび10%(v/v)のウシ胎児血清を補充したイーグル最小必須培地で細胞生存を維持した。サンプルの処理に先立ち、細胞濃度1×105(細胞/ウェル)の培地中で24時間、B16メラノーマ細胞をインキュベートした。 [Inhibition of melamine production]
In order to evaluate the melanin inhibitory effect of 18 types of triterpenoids extracted from loquat leaves, a test using mouse B16 melanoma cells was performed. Mouse B16 melanoma cells were obtained from the RIKEN BioResource Center cell bank. Cell survival was maintained in Eagle's minimum essential medium supplemented with 0.09 mg / ml theophylline and 10% (v / v) fetal calf serum. Prior to sample processing, B16 melanoma cells were incubated for 24 hours in medium at a cell concentration of 1 × 10 5 (cells / well).
びわの葉から抽出された18種類のトリテルペノイドのメラニン抑制効果を評価するため、マウスのB16メラノーマ細胞を用いた試験を行った。マウスのB16メラノーマ細胞は、理研バイオリソースセンター細胞バンクから入手した。濃度0.09mg/mlのテオフィリンおよび10%(v/v)のウシ胎児血清を補充したイーグル最小必須培地で細胞生存を維持した。サンプルの処理に先立ち、細胞濃度1×105(細胞/ウェル)の培地中で24時間、B16メラノーマ細胞をインキュベートした。 [Inhibition of melamine production]
In order to evaluate the melanin inhibitory effect of 18 types of triterpenoids extracted from loquat leaves, a test using mouse B16 melanoma cells was performed. Mouse B16 melanoma cells were obtained from the RIKEN BioResource Center cell bank. Cell survival was maintained in Eagle's minimum essential medium supplemented with 0.09 mg / ml theophylline and 10% (v / v) fetal calf serum. Prior to sample processing, B16 melanoma cells were incubated for 24 hours in medium at a cell concentration of 1 × 10 5 (cells / well).
24時間のインキュベートの後、ウェル当たりの濃度を0.2%未満としたジメチルスルホキシド(DMSO)中に試験サンプルを溶解させ、各ウェルに添加した。サンプル数は3(n=3)である。アルブチンを対照サンプルとして使用した。
After incubation for 24 hours, the test sample was dissolved in dimethyl sulfoxide (DMSO) with a concentration per well of less than 0.2% and added to each well. The number of samples is 3 (n = 3). Arbutin was used as a control sample.
B16メラノーマ細胞をさらに48時間インキュベートし、その後、培地を再度各試料を含む新鮮な培地と交換した。さらに24時間後、メラニン含有量と細胞生存率をそれぞれ405nmおよび570nmでマイクロプレートリーダーを用いて測定した。細胞生存率は、MTT(3-(4,5-ジメチルチアゾール-2-イル)-2,5ジフェニルブロマイド)法を用いて測定した。
B16 melanoma cells were further incubated for 48 hours, after which the medium was again replaced with fresh medium containing each sample. After another 24 hours, melanin content and cell viability were measured using a microplate reader at 405 nm and 570 nm, respectively. Cell viability was measured using the MTT (3- (4,5-dimethylthiazol-2-yl) -2,5 diphenyl bromide) method.
このようにして様々な濃度について、びわの葉から抽出された16種類の化合物のメラニン合成に対する阻害効果および細胞生存性の細胞傷害性効果を試験した。生存細胞の50%に死を引き起こす細胞毒性濃度CC50の値、および、B16メラノーマ細胞のメラニン合成の50%阻害を引き起こす濃度IC50の値を、対応する照射量反応曲線から計算した。CC50およびIC50の値を表1に示す。
Thus, for various concentrations, the inhibitory effect on melanin synthesis and the cytotoxic effect on cell viability of 16 compounds extracted from loquat leaves were tested. The value of the cytotoxic concentration CC 50 causing death in 50% of viable cells and the concentration IC 50 causing 50% inhibition of melanin synthesis in B16 melanoma cells were calculated from the corresponding dose response curves. The values for CC 50 and IC 50 are shown in Table 1.
細胞死の誘導に対するメラニン生成の阻害に関する化合物の相対的有効性、すなわち、CC50の値/IC50の値を選択指数(S.I.:Selectivity Index)として定義し、表1に併せて示した。メラニン合成が阻害される状況としては、メラニン生成が単に阻害される状況の他、B16メラノーマ細胞が死に至る状況がある。CC50は、B16メラノーマ細胞が死に至る、すなわち、細胞傷害性効果の大きさを示したものである。IC50は、細胞傷害性効果を含むメラニン合成に対する阻害効果の大きさを示したものである。メラニン抑制効果の観点から、選択的にメラニン生成を阻害する、すなわち、より高い濃度でなければ細胞傷害性が生じず、より低い濃度であってもメラニン生成を阻害する、高い選択指数(S.I.>1)を有することが好ましい。
The relative efficacy of compounds with respect to the inhibition of melanogenesis against the induction of cell death, ie the value of CC 50 / IC 50 is defined as the selectivity index (SI) and is shown together in Table 1. It was. Situations where melanin synthesis is inhibited include situations where B16 melanoma cells die, in addition to situations where melanin production is simply inhibited. CC 50 indicates that the B16 melanoma cells die, that is, the magnitude of the cytotoxic effect. IC 50 shows the magnitude of the inhibitory effect on melanin synthesis including cytotoxic effects. From the viewpoint of the melanin-suppressing effect, it selectively inhibits melanin production, that is, it does not cause cytotoxicity unless it is at a higher concentration, and inhibits melanin production even at a lower concentration (S. I.> 1).
表1に示す評価結果によれば、8種類のトリテルペノイド、すなわち、化合物1、2、4、5、6、7、8、11は、低い細胞毒性を持ちつつ、高いメラニン生成阻害特性を有する(S.I.>1)ことがわかった。特に、化合物7のマスリン酸は、S.I.の値が1.9と最高であった。この値は、美白成分として知られるアルブチンと同程度である。
According to the evaluation results shown in Table 1, eight types of triterpenoids, that is, compounds 1, 2, 4, 5, 6, 7, 8, and 11 have high melanin production inhibiting properties while having low cytotoxicity ( S.I.> 1) was found. In particular, the maslinic acid of compound 7 is S.I. I. The value of was 1.9, the highest. This value is comparable to that of arbutin known as a whitening component.
活性な化合物の化学構造の不活性な化合物との比較により、以下のことがわかる。ベータ型水酸基をアルファ型水酸基と置き換えた化合物9、12、ケトンと置き換えた化合物13、14、15、および、他の置換基で置き換えた化合物17、18の活性が減少していることが認められることから、C-3位のベータ型水酸基は、メラニン生成阻害活性に重要な役割を果たしている。また、ベータ型の水酸基が存在する場合、化合物(3)は、C-17位のCOOHがCH2OHに還元されることのみにより活性を失うことから、カルボキシル基は、部分的に活性に寄与している可能性がある。化合物5、6に見られるように、カルボキシル基のメチルエステル化は、活性をわずかに低下させる。
Comparison of the chemical structure of the active compound with the inactive compound reveals the following. It can be seen that the activity of compounds 9 and 12 in which the beta-type hydroxyl group is replaced with the alpha-type hydroxyl group, compounds 13 and 14 and 15 in which the ketone is replaced, and compounds 17 and 18 in which the other hydroxyl group is replaced is reduced. Therefore, the beta hydroxyl group at the C-3 position plays an important role in the melanogenesis inhibitory activity. In addition, when a beta-type hydroxyl group is present, the compound (3) loses its activity only by reducing COOH at the C-17 position to CH 2 OH, so that the carboxyl group partially contributes to activity. There is a possibility. As seen in compounds 5 and 6, methyl esterification of the carboxyl group slightly reduces the activity.
このように、びわの葉から抽出されるトリテルペノイドの一部、すなわち、化合物1、2、4、5、6、7、8、11は、メラニン生成を阻害することがわかる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、トルメン酸からなる群から選択される化合物、またはその塩、溶媒和物または水和物は、自然界起源の安全なメラニン抑制剤として用いることができる。
Thus, it can be seen that some of the triterpenoids extracted from loquat leaves, that is, compounds 1, 2, 4, 5, 6, 7, 8, and 11 inhibit melanin production. That is, among triterpenoids extracted from loquat leaves, a compound selected from the group consisting of ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolic acid, maslinic acid, corosolic acid, tolmenic acid, or a salt thereof Solvates or hydrates can be used as safe melanin inhibitors of natural origin.
人の肌が紫外線に過度に晒されるとメラニンおよび異常な色素沈着が過剰に生成される。びわの葉から抽出されるトリテルペノイドの一部は、メラニン生成抑制効果を有するため、肌の美白に対して効果がある。また、びわの葉から抽出されるトリテルペノイドの一部によって、反応性炎症、シミ、肝斑、そばかすなどの様々な皮膚の損傷につながる反応種の過剰生成を抑制することができる。したがって、びわの葉から抽出されるトリテルペノイド、特に、ウルソル酸、オレアノール酸、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、トルメン酸はメラニン生成抑制すなわち美白の機能を有するスキンケア用剤として用いることができる。
• Excessive melanin and abnormal pigmentation are generated when human skin is exposed to UV rays. Some of the triterpenoids extracted from loquat leaves have an effect of suppressing melanin production, and are therefore effective for skin whitening. In addition, some of the triterpenoids extracted from loquat leaves can suppress the excessive generation of reactive species that lead to various skin damages such as reactive inflammation, spots, liver spots, and freckles. Therefore, triterpenoids extracted from loquat leaves, especially ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolic acid, maslinic acid, corosolic acid, and tolmenic acid, are used for skin care with a function of inhibiting melanin production, that is, whitening. It can be used as an agent.
[抗菌]
びわの葉から抽出された18種類のトリテルペノイドの抗菌特性評価用の試験菌としては、プロピオニバクテリウム・アクネスを用いた。試験菌の単一コロニーを採取し、培地に加え、37℃で30時間培養した。次いで、これを細菌懸濁液に加え、105CFU/mlの細菌濃度に調製した。 [antibacterial]
Propionibacterium acnes was used as a test bacterium for evaluating the antibacterial properties of 18 types of triterpenoids extracted from loquat leaves. A single colony of the test bacterium was collected, added to the medium, and cultured at 37 ° C. for 30 hours. This was then added to the bacterial suspension and adjusted to a bacterial concentration of 10 5 CFU / ml.
びわの葉から抽出された18種類のトリテルペノイドの抗菌特性評価用の試験菌としては、プロピオニバクテリウム・アクネスを用いた。試験菌の単一コロニーを採取し、培地に加え、37℃で30時間培養した。次いで、これを細菌懸濁液に加え、105CFU/mlの細菌濃度に調製した。 [antibacterial]
Propionibacterium acnes was used as a test bacterium for evaluating the antibacterial properties of 18 types of triterpenoids extracted from loquat leaves. A single colony of the test bacterium was collected, added to the medium, and cultured at 37 ° C. for 30 hours. This was then added to the bacterial suspension and adjusted to a bacterial concentration of 10 5 CFU / ml.
各化合物は、DMSOに溶解した。ソルビン酸を対照サンプルとして使用した。さらに、DMSOに溶解した各化合物を37℃で30時間インキュベートした。
Each compound was dissolved in DMSO. Sorbic acid was used as a control sample. Furthermore, each compound dissolved in DMSO was incubated at 37 ° C. for 30 hours.
その後、細菌増殖を波長630nmでマイクロプレートリーダーで測定した。最小阻止濃度(MIC)および最小殺菌濃度(MBC)は標準的なブロス微量希釈法により評価した。
Thereafter, bacterial growth was measured with a microplate reader at a wavelength of 630 nm. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were assessed by standard broth microdilution methods.
表2は、化合物濃度が50μg/mlまたは100μg/mlにおけるプロピオニバクテリウム属アクネ菌に対する18種類のトリテルペノイドの抗菌活性を示す。いずれのデータも平均値と標準偏差(n=3)を示した。表2には、活性な化合物の最小阻止濃度(MIC)および最小殺菌濃度(MBC)を併せて示した。対照サンプルであるソルビン酸の抗菌活性は、濃度が400μg/mlのときに、104.4±2.5(%)であった。
Table 2 shows the antibacterial activity of 18 types of triterpenoids against Propionibacterium acne at a compound concentration of 50 μg / ml or 100 μg / ml. All the data showed an average value and a standard deviation (n = 3). Table 2 also shows the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the active compounds. The antibacterial activity of sorbic acid as a control sample was 104.4 ± 2.5 (%) when the concentration was 400 μg / ml.
12種類の化合物、すなわち、化合物1、2、6、7、8、10、11、12、14、16、17、および、18は、それらの最大溶解度でプロピオニバクテリウム属アクネ菌に対して活性を有する。化合物1、7、8および12については、MICが50μg/ml、MBCがそれぞれ100μg/ml、150μg/ml、100μg/ml、および、50μg/mlにおいて、プロピオニバクテリウム属アクネ菌に対する高い活性を示した。なお、トリテルペノイドは、5から8までの範囲のlogP値を有する低い親水性を示す。一般的に、親水性が低い場合には抗菌活性も低い傾向があるが、化合物1、2、6、7、8、10、11、12、14、16、17、および、18は、高い抗菌活性を示している。
Twelve kinds of compounds, namely compounds 1, 2, 6, 7, 8, 10, 11, 12, 14, 16, 17, and 18, are at their maximum solubility against Propionibacterium acne. Has activity. Compounds 1, 7, 8 and 12 show high activity against Propionibacterium acne at MIC of 50 μg / ml and MBC of 100 μg / ml, 150 μg / ml, 100 μg / ml and 50 μg / ml, respectively. Indicated. Triterpenoids exhibit low hydrophilicity with log P values ranging from 5 to 8. In general, antibacterial activity tends to be low when the hydrophilicity is low, but compounds 1, 2, 6, 7, 8, 10, 11, 12, 14, 16, 17, and 18 have high antibacterial activity. Shows activity.
このように、びわの葉から抽出されるトリテルペノイドの一部、すなわち、化合物1、2、6、7、8、10、11、12、14、16、17、18は、プロピオニバクテリウム属アクネ菌に対する抗菌活性を持つことがわかる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、ヒプタジエニック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸からなる群から選択される化合物、またはその塩、溶媒和物または水和物は、自然界起源の安全な抗菌剤として用いることができる。
Thus, some of the triterpenoids extracted from loquat leaves, that is, compounds 1, 2, 6, 7, 8, 10, 11, 12, 14, 16, 17, 18 are propionibacterium acne. It can be seen that it has antibacterial activity against bacteria. That is, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, tolmenic acid, oiscafic acid, 6β, 19-dihydroxyurs-12-en-3 A compound selected from the group consisting of -oxo-28-oic acid, hypotadienic acid, 3-O-trans-p-coumaroyl tormic acid, 3-O-cis-p-coumaroyl tormentic acid, or The salt, solvate or hydrate can be used as a safe antibacterial agent of natural origin.
プロピオニバクテリウム属アクネ菌(P.アクネ)は、毛包脂腺ユニットに主に位置する嫌気性グラム陽性細菌であり、にきびの発症に重要な役割を果たしている。したがって、びわの葉から抽出されるトリテルペノイドの一部は、にきびの発症の抑制に効果がある。よって、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、ヒプタジエニック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸は、にきびの発症を抑制する抗菌剤としての機能を有するスキンケア用剤として用いることができる。
Propionibacterium acne (P. acne) is an anaerobic gram-positive bacterium mainly located in the pilosebaceous unit and plays an important role in the development of acne. Therefore, some of the triterpenoids extracted from leaf leaves are effective in suppressing the development of acne. Therefore, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomol acid, tolmenic acid, oiscafic acid, 6β, 19-dihydroxyurs-12-en-3 -Oxo-28-Oic acid, Hyptadienic acid, 3-O-trans-p-coumaroyl tormic acid, 3-O-cis-p-coumaroyl tormentic acid are antibacterial agents that suppress the development of acne It can be used as a skin care agent having the function of
[抗アレルギー]
びわの葉から抽出された18種類のトリテルペノイドの抗アレルギー効果を評価するため、IgE刺激されたラット好塩基球性白血病(RBL-2H3)細胞からのβヘキソサミニダーゼの放出をチェックした。RBL-2H3細胞は理研バイオリソースセンター細胞バンクから入手した。 [Anti-allergy]
To evaluate the antiallergic effects of 18 triterpenoids extracted from loquat leaves, the release of β-hexosaminidase from IgE-stimulated rat basophilic leukemia (RBL-2H3) cells was checked. RBL-2H3 cells were obtained from the RIKEN BioResource Center cell bank.
びわの葉から抽出された18種類のトリテルペノイドの抗アレルギー効果を評価するため、IgE刺激されたラット好塩基球性白血病(RBL-2H3)細胞からのβヘキソサミニダーゼの放出をチェックした。RBL-2H3細胞は理研バイオリソースセンター細胞バンクから入手した。 [Anti-allergy]
To evaluate the antiallergic effects of 18 triterpenoids extracted from loquat leaves, the release of β-hexosaminidase from IgE-stimulated rat basophilic leukemia (RBL-2H3) cells was checked. RBL-2H3 cells were obtained from the RIKEN BioResource Center cell bank.
RBL-2H3細胞をCO2濃度が5%の加湿空気インキュベーター中、37℃で、10%のFBSおよび1%の抗生物質-抗真菌剤を含むEMEM培地中で維持した。細胞濃度は、2×105細胞/ウェルとした。24時間のインキュベーションの後、培地に1mlの0.5μg/抗DNP(ジニトロフェノール)のマウス免疫グロブリンE(IgEを補充した。さらにこの培養菌を24時間インキュベートした後、培地を、100μlの甲状腺緩衝液で置き換えた。
RBL-2H3 cells were maintained in EMEM medium containing 10% FBS and 1% antibiotic-antimycotic at 37 ° C. in a humidified air incubator with a CO 2 concentration of 5%. The cell concentration was 2 × 10 5 cells / well. After 24 hours of incubation, the medium was supplemented with 1 ml of 0.5 μg / anti-DNP (dinitrophenol) mouse immunoglobulin E (IgE. After further incubation of the culture for 24 hours, the medium was added with 100 μl of thyroid buffer. Replaced with liquid.
0.5%未満の濃度の1μlのDMSOにサンプルを溶解させ、さらに細胞プレートを30分間再度インキュベートした。次いで、甲状腺緩衝液を含むサンプルを、50μg/mlのDNP-BSAを含有する甲状腺緩衝液の100μlで置換した。1時間のインキュベーションの後に、上清の50μlを各ウェルから回収し、それぞれを2Mの基質溶液(p-nitrophenyl-N-acetyl-β-glucosaminide)50μlと混合した。
The sample was dissolved in 1 μl DMSO at a concentration of less than 0.5%, and the cell plate was further incubated for 30 minutes. The sample containing thyroid buffer was then replaced with 100 μl of thyroid buffer containing 50 μg / ml DNP-BSA. After 1 hour incubation, 50 μl of the supernatant was collected from each well and each was mixed with 50 μl of 2M substrate solution (p-nitrophenyl-N-acetyl-β-glucosaminide).
室温において、混合物をシェーカー上で3時間インキュベートした。最後に、100mMの炭酸水素ナトリウムを停止液として100μlを添加した。その後、吸光度を405nmで読み取った。前述と同様のMTT試験により、細胞生存率を測定した。化合物濃度は25μg/mlとした。クェルセチンを対照サンプルとして使用した。
The mixture was incubated for 3 hours on a shaker at room temperature. Finally, 100 μl was added using 100 mM sodium bicarbonate as a stop solution. Thereafter, the absorbance was read at 405 nm. Cell viability was measured by the same MTT test as described above. The compound concentration was 25 μg / ml. Quercetin was used as a control sample.
細胞生存率および抗アレルギー活性の試験結果を表3に示す。いずれのデータも平均値と標準偏差(n=3)を示した。対照サンプルであるクェルセチンの抗アレルギー活性は、72.5±3.0(%)であった。表3における「*」はスチューデントt検定により危険率0.05で対照サンプルとの有意差が見られた化合物、「**」はスチューデントt検定により危険率0.01で対照サンプルとの有意差が見られた化合物を示している。
Table 3 shows the test results of cell viability and antiallergic activity. All the data showed an average value and a standard deviation (n = 3). The antiallergic activity of quercetin as a control sample was 72.5 ± 3.0 (%). “*” In Table 3 is a compound in which a significant difference from the control sample was observed with a student t test at a risk factor of 0.05, and “**” was a significant difference from a control sample in a student t test with a risk factor of 0.01. Indicates a compound in which.
表3に示すように、3種類のトリテルペノイド、すなわち、化合物1、9、および、12は、非細胞毒性濃度においてβ-ヘキソサミニダーゼの放出を大幅に抑制した。化合物7は、わずかな細胞傷害性を持つものの66%の抑制率を示した。
As shown in Table 3, the three types of triterpenoids, ie, compounds 1, 9, and 12, significantly suppressed the release of β-hexosaminidase at non-cytotoxic concentrations. Compound 7 had 66% inhibition although it had slight cytotoxicity.
さらに、化合物1、7、9、および、12について、ヘキソサミニダーゼ放出阻害を引き起こす濃度IC50値を測定し、それぞれ39.5μM、22.86μM、25.6μM、および、15.8μMであることを見出した。
In addition, for compounds 1, 7, 9, and 12, the concentration IC 50 values that cause hexosaminidase release inhibition are measured and are 39.5 μM, 22.86 μM, 25.6 μM, and 15.8 μM, respectively. I found out.
化合物1、7、9、および、12は、12-O-テトラデカノイルホルボール-13-アセテート(TPA:12-O-tetradecanoylphorbol-13-acetate)がマウスに引き起こす炎症、および、TPA誘導性のエプスタイン-バーウイルス早期抗原(EBV-EA)活性化に対して大幅な阻害活性を有することがわかる。
Compounds 1, 7, 9, and 12 are those that cause inflammation caused by 12-O-tetradecanoylphorbol-13-acetate (TPA) in mice and TPA-induced It can be seen that it has significant inhibitory activity against the activation of Epstein-Barr virus early antigen (EBV-EA).
このように、びわの葉から抽出されるトリテルペノイドの一部、すなわち、化合物1、7、9、12は、抗アレルギー性を持つことがわかる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、マスリン酸、3-エピコロソリン酸、オイスカフ酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物は、自然界起源の安全な抗アレルギー剤として用いることができる。
Thus, it can be seen that some of the triterpenoids extracted from loquat leaves, that is, compounds 1, 7, 9, and 12 have anti-allergic properties. That is, among triterpenoids extracted from loquat leaves, a compound selected from the group consisting of ursolic acid, maslinic acid, 3-epicorosolic acid, oiscafic acid, or a salt, solvate or hydrate thereof, It can be used as a safe antiallergic agent of natural origin.
アレルギーや炎症は、体(皮膚、鼻、肺、消化管等)の様々な標的臓器での環境物質に対する過剰反応として現れる。皮膚は、環境との間の主要なインタフェースであるため、不適切な免疫活性は、炎症、皮膚発疹、かゆみや湿疹などとして発症する。したがって、びわの葉から抽出されるトリテルペノイドの一部は、抗アレルギー、抗炎症に効果がある。よって、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、マスリン酸、3-エピコロソリン酸、オイスカフ酸は、抗アレルギーや抗炎症の機能を有するスキンケア用剤として用いることができる。
Allergies and inflammation appear as excessive reactions to environmental substances in various target organs of the body (skin, nose, lungs, digestive tract, etc.). Since the skin is the main interface between the environment, inappropriate immune activity develops as inflammation, skin rash, itching and eczema. Therefore, some of the triterpenoids extracted from loquat leaves are effective for anti-allergy and anti-inflammation. Therefore, among triterpenoids extracted from loquat leaves, ursolic acid, maslinic acid, 3-epicorosolic acid, and oiscafic acid can be used as skin care agents having antiallergic and antiinflammatory functions.
[コラーゲン生成促成]
びわの葉から抽出された18種類のトリテルペノイドのコラーゲン生成促成効果を評価するため、成人正常ヒト皮膚線維芽細胞を用いた試験を行った。成人正常ヒト皮膚線維芽細胞(NHDF-AD)は、Lonza社から入手した。 [Collagen production promotion]
In order to evaluate the collagen formation promoting effect of 18 kinds of triterpenoids extracted from loquat leaves, a test using adult normal human skin fibroblasts was performed. Adult normal human dermal fibroblasts (NHDF-AD) were obtained from Lonza.
びわの葉から抽出された18種類のトリテルペノイドのコラーゲン生成促成効果を評価するため、成人正常ヒト皮膚線維芽細胞を用いた試験を行った。成人正常ヒト皮膚線維芽細胞(NHDF-AD)は、Lonza社から入手した。 [Collagen production promotion]
In order to evaluate the collagen formation promoting effect of 18 kinds of triterpenoids extracted from loquat leaves, a test using adult normal human skin fibroblasts was performed. Adult normal human dermal fibroblasts (NHDF-AD) were obtained from Lonza.
成人正常ヒト皮膚線維芽細胞を10%のFBSおよび1%の抗生物質-抗真菌剤を含むダルベッコ改変イーグル培地(DMEM)中で維持した。培養細胞が接着面いっぱいに広がった状態(コンフルエンス)に成長した後、細胞密度は、5×103/ウェルとした。
Adult normal human skin fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) containing 10% FBS and 1% antibiotic-antimycotic. The cell density was 5 × 10 3 / well after the cultured cells grew to a state where the entire adhesion surface was spread (confluence).
24時間のインキュベーションの後、培地を、サンプル溶液1μlおよび0.5%FBSを補充したDMEMの99マイクロリットルの混合物で置換した。DMSO濃度は、培地中0.5%とした。培養の72時間後、培地中のI型コラーゲン生成量をACEL社のELISAキット用いて測定した。コラーゲン産生量は、同じELISAキットで測定することにより作成した標準曲線を用いて計算した。残りの細胞はMTT評価に用いた。100μMのアスコルビン酸を対照サンプルとして使用した。
After 24 hours of incubation, the medium was replaced with a 99 microliter mixture of DMEM supplemented with 1 μl of sample solution and 0.5% FBS. The DMSO concentration was 0.5% in the medium. After 72 hours of culture, the amount of type I collagen produced in the medium was measured using an ELISA kit manufactured by ACEL. Collagen production was calculated using a standard curve created by measuring with the same ELISA kit. The remaining cells were used for MTT evaluation. 100 μM ascorbic acid was used as a control sample.
それぞれの化合物について、1、5、および、10μg/mlという3種類の化合物濃度におけるコラーゲン生成活性(CP)および細胞生存率(CV)をそれぞれ求めた。コラーゲン生成活性(CP)および細胞生存率(CV)の値は、対照サンプルの場合に対する割合(%)として求めた。次に、細胞生存率(%)あたりの平均のコラーゲン生成活性(%)の平均を表すCP/CV比(>1)を、細胞当たりのコラーゲン生成の刺激活性の指標として求めた。
For each compound, collagen production activity (CP) and cell viability (CV) at three compound concentrations of 1, 5, and 10 μg / ml were determined. Collagen producing activity (CP) and cell viability (CV) values were determined as a percentage of the control sample. Next, a CP / CV ratio (> 1) representing the average of collagen production activity (%) average per cell viability (%) was determined as an index of the stimulation activity of collagen production per cell.
試験結果のコラーゲン生成刺激活性値(CP/CV比)を表4および図1に示す。
The collagen production stimulating activity values (CP / CV ratio) of the test results are shown in Table 4 and FIG.
表4および図1に示すように、いくつかのトリテルペノイドは、細胞あたりの皮膚線維芽細胞のコラーゲン生成を著しく増大させる。びわの葉から抽出される18種類のトリテルペノイドのうち、化合物1、2、3、4、6、7、8、10、13、14の10種類は10μg/mlの濃度でI型コラーゲンを生成した。化合物6、8は、対照サンプルよりも高い、3.4および3.6という特に強力な刺激活性を示している。他の化合物は、中等度の促進効果を示している。
As shown in Table 4 and FIG. 1, some triterpenoids significantly increase collagen production of dermal fibroblasts per cell. Of 18 types of triterpenoids extracted from loquat leaves, 10 types of compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, and 14 produced type I collagen at a concentration of 10 μg / ml. . Compounds 6, 8 show particularly strong stimulating activity of 3.4 and 3.6, which is higher than the control sample. Other compounds show a moderate promoting effect.
びわの葉から抽出される18種類のトリテルペノイドのうち、化合物1、3、4、5、6、7、8、9、10、11、12、13、14、15、17、18の16種類の化合物は、5μg/mlの濃度で、コラーゲン生成の増大を引き起こしている。化合物1、4については、対照サンプルよりも高い、3.8および3.0という値を示している。
Among 18 types of triterpenoids extracted from loquat leaves, 16 types of compounds 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18 The compound causes an increase in collagen production at a concentration of 5 μg / ml. Compounds 1 and 4 show higher values of 3.8 and 3.0 than the control sample.
びわの葉から抽出される18種類のトリテルペノイドのうち、化合物4、8、9、10、13、14の6種類の化合物については、1μg/mlという低い濃度においてもコラーゲン生成活性を有している。
Of the 18 types of triterpenoids extracted from loquat leaves, 6 types of compounds 4, 8, 9, 10, 13, and 14 have collagen producing activity even at a low concentration of 1 μg / ml. .
このように、びわの葉から抽出されるトリテルペノイドの一部、すなわち、化合物1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、17、18は、コラーゲン生成促成特性を持つことがわかる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、3-エピコロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、3-オキソウルス-12-エン-28-オイック酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物は、自然界起源の安全なコラーゲン生成促成剤として用いることができる。
Thus, some of the triterpenoids extracted from loquat leaves, ie, compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17 , 18 have collagen generation-promoting properties. In other words, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl masphosphate, methyl corosolic acid, maslinic acid, corosolic acid, 3-epicorosolic acid, pomolic acid, tolmenic acid, Oiskafic acid, 3-oxourus-12-en-28-oic acid, 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid, 2α, 19α-dihydroxy-3-oxo-12-ursen- A compound selected from the group consisting of 28-oic acid, 3-O-trans-p-coumaroyl tormic acid, 3-O-cis-p-coumaroyl tormentic acid, or a salt or solvate thereof The product or hydrate can be used as a safe collagen production promoter of natural origin.
高齢ヒトの皮膚は、コラーゲン、エラスチン、ヒアルロン酸などの皮膚細胞外マトリックスの著しい減少により、真皮-表皮接合部が平坦化し、機械的張力および弾性が減少する。びわの葉から抽出されるトリテルペノイドの一部は、コラーゲン生成に効果があり、その結果、皮膚のしわの発生を抑制することができる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、3-エピコロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、3-オキソウルス-12-エン-28-オイック酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸、はコラーゲン生成を促成し肌の老化を抑制するコラーゲン生成促成剤の機能を有するスキンケア用剤として用いることができる。
In the elderly human skin, the dermal-epidermal junction is flattened and mechanical tension and elasticity are reduced due to a marked decrease in the extracellular matrix of skin such as collagen, elastin and hyaluronic acid. Some of the triterpenoids extracted from loquat leaves are effective in producing collagen, and as a result, the generation of wrinkles in the skin can be suppressed. In other words, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl masphosphate, methyl corosolic acid, maslinic acid, corosolic acid, 3-epicorosolic acid, pomolic acid, tolmenic acid, Oiskafic acid, 3-oxourus-12-en-28-oic acid, 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid, 2α, 19α-dihydroxy-3-oxo-12-ursen- 28-Oic acid, 3-O-trans-p-coumaroyl tormentic acid, 3-O-cis-p-coumaroyl tormentic acid, stimulates collagen production and suppresses skin aging It can be used as a skin care agent having the function of an agent.
[ヒアルロン酸生成]
びわの葉から抽出された18種類のトリテルペノイドのヒアルロン酸生成促進効果を評価するため、成人正常ヒト皮膚線維芽細胞を用いた試験を行った。成人正常ヒト皮膚線維芽細胞(NHDF-AD)は、Lonza社から入手した。 [Hyaluronic acid production]
In order to evaluate the hyaluronic acid production promoting effect of 18 kinds of triterpenoids extracted from loquat leaves, a test using adult normal human skin fibroblasts was performed. Adult normal human dermal fibroblasts (NHDF-AD) were obtained from Lonza.
びわの葉から抽出された18種類のトリテルペノイドのヒアルロン酸生成促進効果を評価するため、成人正常ヒト皮膚線維芽細胞を用いた試験を行った。成人正常ヒト皮膚線維芽細胞(NHDF-AD)は、Lonza社から入手した。 [Hyaluronic acid production]
In order to evaluate the hyaluronic acid production promoting effect of 18 kinds of triterpenoids extracted from loquat leaves, a test using adult normal human skin fibroblasts was performed. Adult normal human dermal fibroblasts (NHDF-AD) were obtained from Lonza.
成人正常ヒト皮膚線維芽細胞を10%のFBSおよび1%の抗生物質-抗真菌剤を含むダルベッコ改変イーグル培地(DMEM)中で維持した。培養細胞が接着面いっぱいに広がった状態(コンフルエンス)に成長した後、細胞密度は、5×103/ウェルとした。
Adult normal human skin fibroblasts were maintained in Dulbecco's Modified Eagle Medium (DMEM) containing 10% FBS and 1% antibiotic-antimycotic. The cell density was 5 × 10 3 / well after the cultured cells grew to a state where the entire adhesion surface was spread (confluence).
72時間の培養後、培地中のヒアルロン酸の量は、バイオトレーディングパートナーズ社のQnEヒアルロン酸ELISAキットを用いて測定した。ヒアルロン酸産生量は、定量標準曲線から計算した。細胞生存率は、MTT法により測定された。アスコルビン酸を対照サンプルとして使用した。
それぞれの化合物について、1、5、および、10μg/mlという3種類の化合物濃度におけるHA/CV値をそれぞれ求めた。 After 72 hours of culturing, the amount of hyaluronic acid in the medium was measured using a QnE hyaluronic acid ELISA kit manufactured by Bio Trading Partners. Hyaluronic acid production was calculated from a quantitative standard curve. Cell viability was measured by the MTT method. Ascorbic acid was used as a control sample.
For each compound, HA / CV values at three compound concentrations of 1, 5 and 10 μg / ml were determined.
それぞれの化合物について、1、5、および、10μg/mlという3種類の化合物濃度におけるHA/CV値をそれぞれ求めた。 After 72 hours of culturing, the amount of hyaluronic acid in the medium was measured using a QnE hyaluronic acid ELISA kit manufactured by Bio Trading Partners. Hyaluronic acid production was calculated from a quantitative standard curve. Cell viability was measured by the MTT method. Ascorbic acid was used as a control sample.
For each compound, HA / CV values at three compound concentrations of 1, 5 and 10 μg / ml were determined.
試験結果のHA/CV値を表5および図2に示す。
The HA / CV values of the test results are shown in Table 5 and FIG.
ここで、HA/CV値とは、ヒアルロン酸生成促進率であり、ヒアルロン酸濃度の平均値(%)/細胞生存率の平均値(%)である。ヒアルロン酸濃度(HA)および細胞生存率(CV)の値は、対照サンプルの場合に対する割合(%)として求めた。
Here, the HA / CV value is a hyaluronic acid production promotion rate, and is an average value of hyaluronic acid concentration (%) / average value of cell viability (%). Hyaluronic acid concentration (HA) and cell viability (CV) values were determined as a percentage of the control sample.
表5および図2に示すように、いくつかのトリテルペノイドは、細胞あたりの皮膚線維芽細胞のヒアルロン酸生成を著しく増大させる。びわの葉から抽出される18種類のトリテルペノイドのうち、化合物1、2、3、4、6、7、8、10、13、18の10種類の化合物は、10μg/mlの濃度で、ヒト皮膚線維芽細胞におけるヒアルロン酸生成の強力な刺激効果を発揮していた。化合物8、10は、対照サンプルを超える5.9および5.8という刺激活性値を示した。化合物6も高い刺激活性値を示している。
As shown in Table 5 and FIG. 2, some triterpenoids significantly increase hyaluronic acid production of dermal fibroblasts per cell. Of the 18 types of triterpenoids extracted from loquat leaves, 10 types of compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, 18 have a concentration of 10 μg / ml in human skin. It exerted a strong stimulating effect of hyaluronic acid production in fibroblasts. Compounds 8, 10 showed stimulating activity values of 5.9 and 5.8 over the control sample. Compound 6 also shows a high stimulating activity value.
5μg/mlの濃度では、びわの葉から抽出される18種類のトリテルペノイドのうち、化合物1、3、4、6、7、8の6種類の化合物は、ヒト皮膚線維芽細胞におけるヒアルロン酸生成の刺激効果を発揮していた。化合物1は、比較例を超える4.7という刺激活性値を有している。
At a concentration of 5 μg / ml, among the 18 types of triterpenoids extracted from loquat leaves, 6 types of compounds 1, 3, 4, 6, 7, and 8 are capable of producing hyaluronic acid in human skin fibroblasts. It was stimulating. Compound 1 has a stimulating activity value of 4.7 exceeding the comparative example.
低濃度(1μg/mlで)では、びわの葉から抽出される18種類のトリテルペノイドのうち、化合物4の1種類の化合物がヒアルロン酸生成促進効果を示した。
At a low concentration (at 1 μg / ml), one compound of compound 4 out of 18 triterpenoids extracted from loquat leaves showed hyaluronic acid production promoting effect.
このように、びわの葉から抽出されるトリテルペノイドの一部、すなわち、化合物1、2、3、4、6、7、8、10、13、18は、ヒアルロン酸生成促進特性を持つことがわかる。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、3-オキソウルス-12-エン-28-オイック酸、3-O-シス-p-クマロイルトルメンチック酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物は、自然界起源の安全なヒアルロン酸生成促進剤として用いることができる。
Thus, it can be seen that some of the triterpenoids extracted from loquat leaves, that is, compounds 1, 2, 3, 4, 6, 7, 8, 10, 13, 18 have hyaluronic acid production promoting properties. . That is, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, 3-oxours-12-en-28-oic acid, A compound selected from the group consisting of 3-O-cis-p-coumaroyl tormentic acid, or a salt, solvate or hydrate thereof, should be used as a safe hyaluronic acid production accelerator of natural origin. Can do.
若々しい肌は、水分を多く含んでおり、膨圧、弾力性と柔軟性を維持している。ヒアルロン酸は、水分を保持する高い能力を持ち、皮膚の水分量に大きな影響を与える。びわの葉から抽出されるトリテルペノイドの一部は、コラーゲン生成に効果があり、その結果、若々しい皮膚の維持に効果がある。つまり、びわの葉から抽出されるトリテルペノイドのうち、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、3-オキソウルス-12-エン-28-オイック酸、3-O-シス-p-クマロイルトルメンチック酸、は肌の水分量を高める保湿剤としての機能を有するスキンケア用剤として用いることができる。
The youthful skin contains a lot of moisture and maintains swell pressure, elasticity and flexibility. Hyaluronic acid has a high ability to retain moisture and greatly affects the amount of moisture in the skin. Some of the triterpenoids extracted from loquat leaves are effective in producing collagen and, as a result, are effective in maintaining youthful skin. That is, among the triterpenoids extracted from loquat leaves, ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, 3-oxours-12-en-28-oic acid, 3-O-cis-p-coumaroyl tormentic acid can be used as a skin care agent having a function as a moisturizing agent for increasing the moisture content of the skin.
Claims (10)
- びわの葉から抽出されるトリテルペノイドを有効成分として含有するメラニン抑制剤。 Melanin inhibitor containing triterpenoids extracted from loquat leaves as active ingredients.
- 前記びわの葉から抽出されるトリテルペノイドは、ウルソル酸、オレアノール酸、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、トルメン酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物を含むことを特徴とする請求項1に記載のメラニン抑制剤。 The triterpenoid extracted from the leaves of the loquat is a compound selected from the group consisting of ursolic acid, oleanolic acid, betulinic acid, methyl maslinate, methyl corosolic acid, maslinic acid, corosolic acid, tolmenic acid, or a salt thereof, The melanin inhibitor according to claim 1, comprising a solvate or a hydrate.
- びわの葉から抽出されるトリテルペノイドを有効成分として含有する抗菌剤。 An antibacterial agent containing triterpenoids extracted from loquat leaves as active ingredients.
- 前記びわの葉から抽出されるトリテルペノイドは、ウルソル酸、オレアノール酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、ヒプタジエニック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸からなる群から選択される化合物、またはその塩、溶媒和物または水和物を含むことを特徴とする請求項3に記載の抗菌剤。 Triterpenoids extracted from the leaves of the loquat are ursolic acid, oleanolic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, tolmenic acid, oiscafic acid, 6β, 19-dihydroxyurs-12-en-3-oxo -28-Oic acid, Hiptadienic acid, 3-O-trans-p-coumaroyl tormic acid, compound selected from the group consisting of 3-O-cis-p-coumaroyl tormentic acid, or a salt thereof The antibacterial agent according to claim 3, comprising a solvate or a hydrate.
- びわの葉から抽出されるトリテルペノイドを有効成分として含有する抗アレルギー剤。 An antiallergic agent containing triterpenoids extracted from loquat leaves as active ingredients.
- 前記びわの葉から抽出されるトリテルペノイドは、ウルソル酸、マスリン酸、3-エピコロソリン酸、オイスカフ酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物を含むことを特徴とする請求項5に記載の抗アレルギー剤。 The triterpenoid extracted from the loquat leaf contains a compound selected from the group consisting of ursolic acid, maslinic acid, 3-epicorosolic acid, oiscafic acid, or a salt, solvate or hydrate thereof. 6. The antiallergic agent according to claim 5,
- びわの葉から抽出されるトリテルペノイドを有効成分として含有するコラーゲン生成促成剤。 Collagen formation promoter containing triterpenoid extracted from loquat leaves as an active ingredient.
- 前記びわの葉から抽出されるトリテルペノイドは、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、マスリン酸メチル、コロソリン酸メチル、マスリン酸、コロソリン酸、3-エピコロソリン酸、ポモル酸、トルメン酸、オイスカフ酸、3-オキソウルス-12-エン-28-オイック酸、6β,19-ジヒドロキシウルス-12-エン-3-オキソ-28-オイック酸、2α,19α-ジヒドロキシ-3-オキソ-12-ウルセン-28-オイック酸、3-O-トランス-p-クマロイルトルメンチック酸、3-O-シス-p-クマロイルトルメンチック酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物を含むことを特徴とする請求項7に記載のコラーゲン生成促成剤。 The triterpenoids extracted from the leaves of the loquat are ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl masphosphate, methyl corosolic acid, maslinic acid, corosolic acid, 3-epicorosolic acid, pomol acid, tolumic acid, oiscafic acid , 3-oxourus-12-ene-28-oic acid, 6β, 19-dihydroxyurs-12-en-3-oxo-28-oic acid, 2α, 19α-dihydroxy-3-oxo-12-ursen-28- Oic acid, 3-O-trans-p-coumaroyl tormentic acid, a compound selected from the group consisting of 3-O-cis-p-coumaroyl tormentic acid, or a salt, solvate thereof or The collagen production promoting agent according to claim 7, comprising a hydrate.
- びわの葉から抽出されるトリテルペノイドを有効成分として含有する保湿剤。 Moisturizer containing triterpenoids extracted from loquat leaves as active ingredients.
- 前記びわの葉から抽出されるトリテルペノイドは、ウルソル酸、オレアノール酸、ウバオール、ベツリン酸、コロソリン酸メチル、マスリン酸、コロソリン酸、ポモル酸、3-オキソウルス-12-エン-28-オイック酸、3-O-シス-p-クマロイルトルメンチック酸、からなる群から選択される化合物、またはその塩、溶媒和物または水和物を含むことを特徴とする請求項9に記載の保湿剤。
Triterpenoids extracted from the leaves of the loquat are ursolic acid, oleanolic acid, ubaol, betulinic acid, methyl corosolic acid, maslinic acid, corosolic acid, pomolic acid, 3-oxours-12-en-28-oic acid, 3- The humectant according to claim 9, comprising a compound selected from the group consisting of O-cis-p-coumaroyl tormentic acid, or a salt, solvate or hydrate thereof.
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CN108478481A (en) * | 2018-04-20 | 2018-09-04 | 珠海市自然之旅生物技术有限公司 | A kind of preservative and its preparation method and application containing Chinese cassia tree |
CN108478481B (en) * | 2018-04-20 | 2022-09-02 | 珠海市自然之旅生物技术有限公司 | Preservative containing cinnamon and preparation method and application thereof |
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