JP4897276B2 - New glyceroglycolipid - Google Patents

Description

  The present invention relates to a novel glyceroglycolipid, a melanin production inhibitor using the same, a cosmetic, and a food.

Due to air pollution and destruction of the ozone layer, the amount of ultraviolet rays that reach the human epidermis tends to increase year by year. Along with this, skin problems such as skin spots, freckles, and darkness due to ultraviolet rays become a major problem. ing. By irradiating with ultraviolet rays, tyrosine present in the skin is oxidized by the action of tyrosinase enzyme to produce melanin pigment, and if this is produced excessively, it causes skin problems such as spots, freckles, and darkness. appear. As a method for inhibiting the production of this melanin pigment and preventing spots, freckles, and darkness, L-ascorbic acid and its derivative L-ascorbic acid glucose distribution have been conventionally applied to skin cosmetics for the purpose of skin whitening. A saccharide is blended (for example, see Patent Document 1), and a skin external preparation formulated by combining L-ascorbic acid glucose glycoside with an amino acid or a derivative thereof (for example, see Patent Document 2). Many have been proposed. However, these whitening agents do not always have a sufficient whitening effect, and if a component showing the whitening effect is blended in a concentration that allows the whitening effect, there may be problems in safety and stability.
On the other hand, the present inventor has been searching for a melanin production inhibitor for the purpose of whitening the skin, but kale which is a cruciferous vegetable or an extract thereof has an action to inhibit melanin production by ultraviolet rays. The patent application has already been filed (see Patent Document 3).

JP-A-4-18212 JP-A-4-182413 JP 2004-91396 A

  An object of the present invention is to provide a novel glyceroglycolipid, a melanin production inhibitor, a cosmetic, and a food containing the novel glyceroglycolipid.

The main configuration of the present invention is as follows.
Melanin production inhibitor characterized by containing a compound represented by the active ingredient (1) Structural Formula (I).

(However, the acyl group of 7,10-hexadecadienoic acid and the acyl group of 9,12,15-octadecatrienoic acid in the formula may be interchanged)
(2) A whitening cosmetic comprising the melanin production inhibitor according to (1) as an active ingredient.

  The glyceroglycolipid of the formula (I) used in the present invention is a novel substance, has a melanin production inhibitory action, and exhibits a whitening effect. A whitening cosmetic containing this glyceroglycolipid can be provided. A food containing the glyceroglycolipid can be provided. It was also confirmed that kale-derived glyceroglycolipid was effective.

Hereinafter, the novel glyceroglycolipid of the present invention, the melanin production inhibitor containing the same, cosmetics, and foods will be described.
First, the glyceroglycolipid of the present invention has the above chemical formula (I) (wherein the 7,10-hexadecadienoic acid acyl group and the 9,12,15-octadecatrienoic acid acyl group may be interchanged). A glyceroglycolipid having a galactose as a saccharide group and an acyl group having 16 and 18 carbon atoms containing an unsaturated bond as an acyl group.

  Specifically, the novel glyceroglycolipid of the present invention is represented by the chemical formula (I) represented by the IUPAC name as follows (wherein the 7,10-hexadecadienoic acid acyl group and 9,12,15 in the formula: -The octadecatrienoic acid acyl group may be interchanged).

IUPAC name of glyceroglycolipid of formula (I)
(9Z, 12Z, 15Z) -1-((7Z, 10Z) -hexadeca-7,10-dienoyloxy) -3-((2R, 3R, 4S, 5R, 6R) -3,4,5-trihydroxy-6 -(hydroxymethyl) -tetrahydro-2H-pyran-2-yloxy) propan-2-yl-octadeca-9,12,15-trienoate
IUPAC name of the glyceroglycolipid in which the acyl group of 7,10-hexadecadienoic acid and the 9,12,15-octadecatrienoic acid acyl group of the chemical formula (I) are interchanged
(7Z, 10Z) -1-((9Z, 12Z, 15Z)-octadeca-9,12,15-trienoyloxy) -3-((2R, 3R, 4S, 5R, 6R) -3,4,5-trihydroxy -6- (hydroxymethyl) -tetrahydro-2H-pyran-2-yloxy) propan-2-yl-hexadeca-7,10-dienoate

  The glyceroglycolipid represented by the chemical formula (I) of the present invention (wherein the acyl group of 7,10-hexadecadienoic acid and the acyl group of 9,12,15-octadecatrienoic acid in the formula may be interchanged) For example, it can be obtained separately from an ethanol extract of kale, which is a vegetable of the cruciferous plant. This kale is a cruciferous vegetable native to Southern Europe and is said to be the original species of cabbage. Its scientific name is Brassicca Oleracea L. var. acephala DC. There are various types of kale such as kitchen kale, mallow kale, bush kale, tree kale, collard, green alga kanran, and any of these can be used to produce the novel glyceroglycolipid of the present invention. The use part of kale for obtaining the novel glyceroglycolipid of the present invention is not limited to those usually used for food such as leaves, but can also use kale stems, flowers, roots, etc. It is not a thing and the cultivation method and cultivation place are not specifically limited.

The glyceroglycolipid represented by the chemical formula (I) of the present invention (wherein the acyl group of 7,10-hexadecadienoic acid and the acyl group of 9,12,15-octadecatrienoic acid in the formula may be interchanged) In the extract obtained by extracting kale leaves, stems, flowers, roots and the like with ethanol having a concentration of 50 to 100% by mass, preferably 70 to 100% by mass, it is contained in a large amount in the fat-soluble fraction. ing.
Specifically, for example, ethanol is added to a dry matter such as kale leaves, stems, flowers and roots at a concentration of 50 to 100% by mass, preferably 70 to 100% by mass, and immersed at room temperature for several hours to several tens of hours. -The extract is obtained by stirring, and the extract is concentrated by heat evaporation or the like. The extract obtained here can be extracted again using an aqueous ethanol solution having a concentration of 70% by mass and dichloromethane, and separated into two layers by a separatory funnel. The obtained dichloromethane fraction can be concentrated by heat evaporation, etc., extracted again with normal hexane and methanol, and separated into two layers by a separatory funnel. The obtained normal hexane layer was concentrated by heating evaporation and the like, and fractionated and purified by silica gel column chromatography and reverse phase HPLC to obtain the chemical formula (I) (7,10-hexadecadienoic acid acyl in the formula). Group and 9,12,15-octadecatrienoic acid acyl group may be interchanged) to obtain a glyceroglycolipid.

  Next, in the melanin production inhibitor of the present invention, the glyceroglycolipid thus obtained may be used as an active ingredient as it is, or the chemical formula (I) (wherein 7, A kale extract containing a glyceroglycolipid represented by 10-hexadecadienoic acid acyl group and 9,12,15-octadecatrienoic acid acyl group may be used as an active ingredient. In the melanin production inhibitor containing the novel glyceroglycolipid of the present invention, any of various components that do not impair the effects of the present invention and do not adversely affect them can be used as auxiliaries, media or carriers.

The novel glyceroglycolipid of the present invention has a skin melanogenesis inhibitory action. Ultraviolet rays oxidize tyrosine present in the skin by the action of the tyrosinase enzyme to produce melanin pigment. If this is excessively produced, it causes skin problems such as spots, freckles, and darkness. By using the novel glyceroglycolipid of the present invention, the production of melanin in the skin when irradiated with ultraviolet rays is suppressed, and such skin troubles are effective for improvement.
A cosmetic or external preparation for skin containing the novel glyceroglycolipid of the present invention can be produced. Such cosmetics include oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, metals Contains sequestering agents, polymers such as water-soluble polymers, thickeners, powder components, UV absorbers, UV blockers, moisturizers such as hyaluronic acid, perfumes, pH adjusters, desiccants, etc. be able to. In addition, vitamins, skin activators, blood circulation promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, anticancer agents, whitening agents, bactericides, and other medicinal and physiologically active ingredients may be included. it can.
Cosmetics include skin lotions such as lotions, emulsions, creams, packs, makeup base lotions, makeup creams, emulsions or creams, or ointment-type foundations, lipsticks, eye colors, cheek colors, etc. Cosmetics for body use such as makeup cosmetics, hand creams, leg creams, body lotions, and the like, and bathing agents, oral cosmetics, hair cosmetics and the like can be used. As the cosmetics containing the novel glyceroglycolipid of the present invention, for example, emulsions, cosmetics, face creams, hand creams, lotions, essences and the like are preferable from the functional aspect.
Such a cosmetic containing the novel glyceroglycolipid of the present invention can be produced according to a conventional method. The addition amount of the novel glyceroglycolipid of the present invention in cosmetics is not particularly limited, but as an example, 0.001 to 20 mass%, preferably 0.01 to 10 mass% of the total cosmetic weight. % Is appropriate.

As a food composition, the novel glyceroglycolipid of the present invention can be used as it is, as a food for health, or as a food material, with various nutritional components added or contained in food or drink. For example, suitable auxiliaries such as starch, lactose, maltose, vegetable oil powder, cocoa butter powder, stearic acid and the like can be added.
As a food composition, it may be formed into an edible form, for example, granular, granular, tablet, capsule, paste, etc. and used for edible, and various foods such as ham, sausage, etc. It may be used by adding it to food, processed fish foods such as kamaboko, chikuwa, bread, confectionery, butter, powdered milk, fermented dairy products, or it may be used by adding to beverages such as water, fruit juice, milk, soft drinks, etc. .
The blending amount of the novel glyceroglycolipid of the present invention into the food composition is appropriately set depending on the type and state of the food composition.

  Next, the present invention will be described in more detail with reference to examples. The following production examples, examples and formulation examples show preferred examples of the present invention and are not limited thereto. Further, “%” in each example is based on mass.

Production of novel glyceroglycolipids The present invention will now be described in more detail by way of examples. The following production examples, examples and formulation examples show preferred examples of the present invention and are not limited thereto. Further, “%” in each example is based on mass.

1) Extraction of Kale fat soluble fraction Crush the dried Kale leaf, add 3L of 100% ethanol to 500g of Kale pulverized product, extract for 1 hour with heating, and concentrate this extract to 1L using an evaporator. did. Further, purified water was added so that the ethanol concentration became 70%, dichloromethane was added, and the mixture was separated with a separatory funnel. Further, the fraction eluted with dichloromethane was concentrated to dryness, dissolved in n-hexane, methanol was added, and the mixture was separated with a separatory funnel. The obtained n-hexane layer was concentrated and dried to obtain 12.4 g of “lipophilic fraction A” of the kale ethanol extract. The fractionation flow is shown in FIG.

2) Separation of fat-soluble fraction A by silica gel column chromatography The obtained fat-soluble fraction A (12.4 g) was used for separation and purification by the following method and conditions.
-Preparative column: Wako silica gel 60 0.040-0.063mm
Inner diameter 6cm, length 60cm
Eluent: Dichloromethane, methanol eluent composition of dichloromethane: methanol = 100: 0 → 100: 1 → 100: 2 → 100: 10 → 5: 1 → 3: 1 → 0: 100 in order to the column Washed away.
・ Preparation
The components were eluted from the eluent composition dichloromethane: methanol = 5: 1 and fractionated into 4 fractions B to E. Among them, the first fraction B eluted with dichloromethane: methanol = 5: 1 was used. 5.8 g of fraction B was obtained.

3) Separation of fraction B by silica gel column chromatography The obtained fraction B (5.8 g) was used for separation and purification by the following method and conditions.
-Preparative column: Wako silica gel 60 0.040-0.063mm
Inner diameter 6cm, length 60cm
Eluent: n-hexane, ethyl acetate, methanol The composition of the eluent is n-hexane: ethyl acetate: methanol = 10: 1: 0 → 5: 1: 0 → 2: 1: 0 → 1: 1: 0 → 1: 1: 0.1 → 1: 1: 0.2 → 1: 2: 0.2 → 1: 2: 0.5 → 0: 1: 1 → 0: 0: 1 Washed away.
・ Preparation
The components were eluted from the eluent of n-hexane: ethyl acetate: methanol = 2: 1: 0, and the first fraction F was obtained. Fractions were fractionated into 8 fractions from fraction F to fraction M. Fraction K eluting with n-hexane: ethyl acetate: methanol = 1: 2: 0.2 → 1: 2: 0.5 was used. 2.0 g of fraction K was obtained.

4) Separation and purification of fraction K by high performance liquid chromatography A portion (160 mg) of fraction K was separated and purified by the following method and conditions.
·Equipment configuration:
Preparative column: SHISEIDO CAPCELL PAK C18 UG-120 10.0 × 250mm
Detector: Shimadzu SPD-M10AVP UV215nm
・ Eluent:
Flow rate: 4.7mL / min
Eluent: Water: acetonitrile = 10: 90
-Fraction Fraction W (9 mg) was obtained from the fraction having a retention time of 26 minutes to 27 minutes. This fraction W is referred to as “sample (a)”.
FIG. 1 shows a flow chart for fractionating a sample (a) from an ethanol extract of kale.

5) Determination of structure of glyceroglycolipid With respect to the sample (a) collected in the above 4), the structure of the compound was determined using an LC / MS analyzer and an NMR analyzer.
LC / MS analyzer, conditions and equipment configuration:
HPLC system: PE Series 200
MS system: PE-Sciex API 150 (+ / (-)-ESI, Fast-Switching-Mode)
Analysis column: Merck Select B 250 × 4 mm, 5 μm
・ Eluent:
Flow rate: 1 mL / min
Eluent: A ... 0.5 mM ammonium formate, 0.1% formic acid aqueous solution
B: 0.5 mM ammonium formate, 0.1% formic acid in acetonitrile: methanol = 1: 1 solution Gradient: shown in Table 1.

Analysis conditions for ¹ H-NMR Measurement was performed at a resonance frequency of 400 MHz. The solvent was deuterated methanol and used for chemical shift standards ( ¹ H-NMR 3.30 ppm).
The results of LC / MS analysis and NMR analysis of each sample extracted and separated are shown below. In Table 2, “Gal” represents galactose and “FA” represents fatty acid.
Sample (a):
The LC / MS and NMR data are shown below.
LC / MS
(+)-ESI: 766 [M + NH ₄ ] ⁺ , 587 [M-Gal + H] ⁺
(−)-ESI: 793 [M + HCOO] ⁻ , 747 [M−H] ⁻ , 277 [C ₁₈ H ₂₉ O ₂ ] ⁻ , 251 [C ₁₆ H ₂₇ O ₂ ] ^−
1 H-NMR
The spectral assignments and chemical shifts are shown in Table 2.

  Sample (a) is a novel glyceroglycolipid represented by chemical formula (I), or a 7,10-hexadecadienoic acid acyl group and 9,12,15-octadecatrienoic acid acyl group in chemical formula (I). It is a new glyceroglycolipid that has been replaced, or a mixture thereof, and shows the spectral data as described above. By LC / MS and NMR analysis, it was not possible to determine the bonding positions of the two types of fatty acyl groups.

Melanin Production Inhibition Evaluation Test Using mouse-derived cultured B16 melanoma cells as cells producing melanin, the cells are cultured in Eagle's MEM medium supplemented with fetal bovine serum to a final concentration of 10%, and the cells are 1 × 10 ^3. Cells / well were seeded in 96-well plates and cultured in a CO ₂ incubator for 5 days. Thereafter, the medium was replaced with the medium to which the test sample was added, and further cultured for 3 days under the same conditions. After completion of the culture, the cells are washed, the cells are solubilized with sodium dodecyl sulfate (SDS), and the absorbance at 475 nm and 260 nm is measured, which are designated as S475 and S260. The melanin inhibition rate was calculated by the following equation using the absorbance at 475 nm and 260 nm of cells cultured in a medium not added with the test sample as C475 and C260.

As the test sample, the chemical formula (I) obtained in Example 1 (wherein the 7,10-hexadecadienoic acid acyl group and the 9,12,15-octadecatrienoic acid acyl group in the formula are interchanged). Sample (a), which is a glyceroglycolipid represented by the formula (1), was added to the medium at 12.5 μg / mL, 25.0 μg / mL, and 50.0 μg / mL, respectively. As a positive control, kojic acid, which is widely used as a whitening component, was used, and 12.5 μg / mL, 25.0 μg / mL, and 50.0 μg / mL were added to the medium.
The results of these melanin production inhibition evaluation tests are shown in FIG. As shown in FIG. 2, the sample (a) of the novel glyceroglycolipid of the present invention showed a melanin production inhibitory effect in the same manner as the positive control kojic acid.

  Next, formulation examples as cosmetics, which are specific usage forms of the novel glyceroglycolipid of the present invention, are shown below.

Formulation Example 1: Toner Lotion Chemical Formula (I) obtained in Example 1 above (wherein 7,10-hexadecadienoic acid acyl group and 9,12,15-octadecatrienoic acid acyl group are interchanged) A lotion having the following composition was prepared by a conventional method using the glyceroglycolipid represented by (sample (a)).

Formulation Example 2: Cream Using the novel glyceroglycolipid (sample (a)) obtained in Example 1 above, a cream having the following composition was produced by a conventional method.

Formulation Example 3: Cosmetic Pack Using the novel glyceroglycolipid (sample (a)) obtained in Example 1 above, a cosmetic pack having the following composition was produced by a conventional method.

Formulation example 4: Melanin production inhibitor (tablet)
Using the novel glyceroglycolipid (sample (a)) obtained in Example 1 above, tablets of the composition for inhibiting melanin production having the following composition were produced by a conventional method.

The flowchart which fractionates a sample (a) from the ethanol extract of a kale. The graph which shows the result of the evaluation test of melanin production suppression of the sample (a) obtained in Example 1. FIG.

Claims (2)

Melanin production inhibitor characterized by containing a compound represented by as an active ingredient in Structural Formula (I).

(However, the acyl group of 7,10-hexadecadienoic acid and the acyl group of 9,12,15-octadecatrienoic acid in the formula may be interchanged) A whitening cosmetic comprising the melanin production inhibitor according to claim 1 as an active ingredient.