JP2003226649A - Agent for preventing and treating insufficiency of melanin production - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a melanin production promoter, a tyrosinase activity promoter and a medicament for preventing and/or treating insufficiency of melanin production such as trichopoliosis and leukoplasia. <P>SOLUTION: The melanin production promoter, the tyrosinase activity promoter and the medicament for preventing and/or treating the insufficiency of the melanin production contain a plant belonging to Piperaceae or an extract thereof. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a prophylactic and / or therapeutic agent for leukoplakia, melanin deficiency diseases such as leukoplakia. More specifically, maintaining black hair,
The present invention relates to a drug having a melanin production promoting action of melanoisite, which produces melanin, which is deeply involved in skin color determination.

[0002]

2. Description of the Related Art Melanin produced by melanin-producing cells existing in the epidermis or hair is transferred to keratinocytes and supplied and dispersed in tissues as the keratinocytes proliferate and differentiate. At this time, the color tone of the tissue is determined by the quantitative and qualitative difference of melanin supplied to the tissue. Pigmentation diseases such as melanosis of the skin caused by spots, freckles, dark skin and drugs such as steroids are diseases caused by excessive deposition of melanin pigment on the skin.

On the other hand, as a result of the lack or reduction of melanin production ability, leukoplakia and vitiligo of the skin occur with age. Leukoplakia is a phenomenon in which white hair is formed in localized islands, and is mainly caused by a decrease in melanin formation due to deficiency of enzymatic activity of tyrosinase in melanocytes in hair follicles. This melanin is widely distributed in the animal and plant kingdoms, but in vertebrates, tyrosine is oxidized by tyrosinase in the cytoplasmic granular melanosomes in melanocytes, and dopa and dopaquinone are biosynthesized. Is known to be converted into indolequinone, and melanin is biosynthesized through a complicated pathway.
Such failure of the melanin production mechanism is unfavorable for beauty regardless of gender.

However, at present, most of the treatment methods for albinism are dyeing exclusively with dyes, and various hair cosmetics have been reported as therapeutic agents, but they are widely used as the fundamental therapeutic agents. There is nothing that has been applied.

Vitiligo is a type of primary eruption and refers to a plaque caused by demelanization of melanin. Vitiligo vulgaris, which has the most cases, is predisposed to systemic modulation of endocrine, autonomic dysfunction, internal organ disease, etc. This is caused by clear demarcation of melanin on a part of the skin. There is currently no suitable treatment for such vitiligo.

[0006]

Therefore, the present invention prevents and / or prevents melanin production deficiency such as leukoplakia and leukoplakia, which is excellent in melanin production promoter, tyrosinase activity promoter, and melanin production promoter effect. It was made for the purpose of providing a drug for treatment.

[0007]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that a specific plant or an extract thereof has an excellent effect of promoting melanin production and suppresses melanin deficiency diseases such as leukoplakia and leukoplakia. The present invention was completed by finding that it can be prevented and / or treated. That is, the present invention is as follows. (1) A melanin production promoter containing a plant belonging to the family Piperaceae or an extract thereof. (2) Pepper genus or peperomia (P
A melanin production promoter containing a plant belonging to the genus eperomia) or an extract thereof. (3) A tyrosinase activity promoter containing a plant belonging to the family Piperaceae or an extract thereof. (4) Pepper genus or peperomia (P
A tyrosinase activity promoter containing a plant belonging to the genus eperomia) or an extract thereof. (5) A preventive and / or therapeutic agent for melanogenesis insufficiency containing a plant belonging to the pepper (Piperaceae) family or an extract thereof. (6) Pepper genus or peperomia (P
An agent for preventing and / or treating melanogenesis deficiency, which comprises a plant belonging to the genus eperomia) or an extract thereof. (7) The above (5) or (6), wherein the dosage form is a skin external preparation
The preventive and / or therapeutic agent for the described melanin deficiency disease.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention relates to pepper (Piper).
aceae) and a plant belonging to the family Aceae) or an extract thereof as an active ingredient. Pepper
In the family Ceae), about 3,000 species of 8 genera are known, and most of them belong to the genus Pepper and the genus Peperomia.

As a plant belonging to the genus Pepper, pepper (Piper niger) is used.
m), betel (Piper betle), beetle vine (Piper kadzura),
Hippopotamus (Piper methysticum), Pepper magnificucu
m), cubbeba (Piper cubeba), hihatsu (Piper retrofractum), and euglena (Piper postelsianu)
m), Guiana Pepper (Piper clusi
i), Piper guienes
e), Naga pepper (Piper longum), Pertata (Piper pelata), Umberlata (Piper umbellata), Javan pepper (Piper retrofractum), etc. can be mentioned. Also, Peperomia
As plants belonging to the genus ia, Aspera (Pepe)
romia asperula), Columella (Pepe)
romia columella), Dorabriformis (Peperomia dolabriformi)
s), Graveolence (Peperomia grav)
eolens), Rotundifolia (Peperom)
iarotundifolia), blue-green algae (Peperomia argyreia), and blue-green algae (Peperomia capera)
ta), sycamore (Peperomia boni)
Nisimensis), Sadasou (Satagusa: Peper)
omia japonica), hedelifolia (Pe
peromia hederifolia), Obtusifolia (Peperomia obtusifolia)
a), Magnolia eforia (Peperomia m)
agnolia efolia), Gravera (Peper)
omia glabella), Pteorata (Pep)
eromia puteolata, scandense (Peperomia scandens), botteri (Peperomia botteri), brewipes (Peperomia brevipes), class siforia (Peperomia crassia)
lia), Ebrunea (Peperomia ebur)
nea), Hederiforiea (Peperomia he
derifolia), incarna (Peperomi)
a incana), Macrossa (Peperomia)
maculosa), Marmolata (Peperom)
ia marmorata), Metallica (Pepper)
omia metallica), Nemmurarifolia (Peperomia nummularifolia)
a), Thundershie (Peperomia sande)
rsii), Titimalloydes (Peperomi)
a tithymaloides), Griseoargentea (Peperomia griseoargene)
tea), Serpense (Peperomia serp)
ens), Blue-bellied algae (Peperomia)
verschaffeltii) and the like. In the present invention, plants belonging to the genus Pepper are preferable, and among them, pepper (Pepper: Pip) is preferable.
er nigrum) and betel (ginger sauce: Piper)
betle) is preferred.

In the present invention, pepper (Pipera)
The plants belonging to the family caeae) can be used as raw or dried, but from the viewpoints of usability, formulation, etc., it is preferable to use as an extract thereof, for example, a dry powder or a solvent extract. . The plant belonging to the pepper (Piperaceae) family of the present invention means one containing all cells constituting this plant, and it is also possible to use all of this plant or a part of the part constituting the plant. The parts that make up the plant are leaves,
Examples thereof include roots, rhizomes, stems, flowers, fruits and pericarps.

In the present invention, pepper (Pipera)
An extract of a plant belonging to the family Ceae) can be obtained by a conventional method. For example, a generally known method for obtaining an extract by extracting leaves, roots, rhizomes, stems, flowers, fruits, pericarps, etc. of plants with water and / or a hydrophilic organic solvent can be used. Furthermore, from this extract, freeze drying, spray drying,
The powder can be obtained by a generally known solvent removal method such as distillation under reduced pressure. Examples of the hydrophilic organic solvent include lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol and butanol, ethers having 2 to 4 carbon atoms such as ethyl ether and methyl ether, and acetone and ethyl methyl ketone. And the ketone of 4. Of these, ethanol is particularly preferable. These solvents may be used alone or in combination of two or more kinds, or water and these hydrophilic organic solvents may be mixed and used. A preferable extraction solvent as the mixed solvent obtained by mixing with water is a hydroalcohol, and hydrous ethanol is particularly preferable. The amount of these extraction solvents used is not particularly limited. The specific method of extraction is
Known methods such as a cold immersion method, a digestion method, and a percolation method, which are used when producing an extract or a tincture, can be applied. The obtained extract may be used as it is, or may be further concentrated, diluted, or purified before use. Furthermore, by purifying these extracts or powders using column chromatography or the like, a single component or a mixture of a plurality of components can be used.

Pepperac according to the present invention
A plant belonging to the family eae) or an extract thereof can be administered orally or parenterally. The dosage form may be selected according to the target disease. The formulation to be administered orally includes tablets, capsules, powders, granules, fine granules,
A dosage form such as an oral solution can be mentioned. Further, as a preparation to be administered parenterally, injections, extracts, alcoholic drugs, suppositories, suspensions, tinctures, ointments, poultices, liniments, lotions, aerosols, plasters, etc. Mention may be made of the external preparation for skin. In the present invention, an external preparation for skin which is easy to administer is preferable. Also,
Plants belonging to the pepper (Piperaceae) family or extracts thereof are lotions, creams, lotions, emulsions, foams, foundations, packs, skin cleansers, shampoos, conditioners, hair tonics, hair liquids, hair creams, etc. You may mix | blend with the cosmetic composition of.

The formulation can be carried out by a known formulation technique, and appropriate formulation additives can be added to the formulation. Formulation additives include excipients, suspending agents, emulsifying agents, moisturizing agents (moisturizing agents), preservatives, surfactants, thickeners,
Examples thereof include dyes, fragrances, propellants, and the like, and formulation additives may be added appropriately as long as the effects of the present invention are not impaired.

The preventive and / or therapeutic agent for melanin production deficiency of the present invention may further contain known tyrosinase activity promoters, melanin production promoters and the like. As these components, for example, salamander, takasaburo, pre-pandaku, saffron, cayu rapé, guacomist,
Algodonera, Huairuru, Angarate, Pinguica, Aritasso, Zapote, Axcopaque, Unshu mandarin, Natsumikan, Orange, Hassaku, Iyokan,
Grapefruit, Yuko, Sudachi, Kabos, Ponkan, Kumquat, Kins, Malkin, Nagakin,
Botanicals such as Karachi, Coptis, Senburi, Ashitaba,
Basidiomycetes, cockle, green mussel, moss, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, oyster mushrooms, moss mushrooms, moss mushrooms, moss mushrooms, moss mushrooms, moss mushrooms, moss mushrooms, moss mushrooms Red shell, abalone,
Examples thereof include shellfish such as turban shells and baits, and extracts thereof. One or two or more of these components may be added to the plant (Piperaceae) belonging to the present invention or an extract thereof.

Pepperacea in the formulation
e) The amount of the plant belonging to the family or the extract thereof may be appropriately examined in consideration of the target disease, sex, age, symptoms, etc., but the dry solid content of the plant belonging to the family Piperaceae is included in the formulation. (In the case of extract, it is 0.0001 to 20 in terms of dry solid content of the plant used for extraction)
It is preferably blended by weight, and more preferably 0.01-10 wt%.

[0016]

EXAMPLES Next, the present invention will be further described with reference to examples, but the present invention is not limited to these examples.

Example 1 Dried leaves and / or stems of pepper and betel were crushed and then crushed with a 5-fold amount of 50% ethanol aqueous solution for 2 hours.
It was extracted twice while hot. The extract was filtered while hot, and the filtrate was concentrated under reduced pressure and freeze-dried to obtain a dry 50% ethanol aqueous solution extract.

Example 2 Melanin production promoting action in melanoma cell culture method The melanin production promoting action of the extract obtained in Example 1 was examined.

Cell culture method: B16 melanoma cultured cells derived from mouse were added with D-containing 10% fetal bovine serum (FBS).
2 × 10 ⁴ ce in MEM medium (Invitrogen)
It was adjusted to lls / well (6 well plate) and precultured in a CO ₂ incubator (5% CO ₂ ) at 37 ° C. for 24 hours. The sample is dimethyl sulfoxide (DMS
O), dissolve in a 1: 1 solution of PBS and add 10% F to this solution.
The concentration was adjusted by diluting with D-MEM medium containing BS.
Final analyte concentrations of 0.1, 1.0 and 10.0 μg
4 ml each was used. In addition,
The final concentration of DMSO was adjusted to 0.1%. As a control, dimethyl sulfoxide (DMSO), PBS 1: 1
4 μl of the same dilution of the solution was used. The culture was further continued for 3 days, and the intracellular and extracellular melanin amounts were measured by the following method.

Measurement of intracellular melanin: Cells collected by trypsin treatment were treated with Ca, Mg Free (CMF) -PBS.
After washing with 1 mol / l NaOH (400 μl)
And dissolve by heating at 80 ° C for 30 minutes, then the absorbance of this solution
The intracellular melanin amount was determined from (475 nm).

Measurement of extracellular melanin: an equal amount of 0.4 mo
1 / HEPES buffer (pH 6.8) / ethanol (9: 1, v / v) was added to adjust the pH to 7.0, and the supernatant of the medium (700 × g, 10 minutes, prepared by centrifugation at 4 ° C.) The amount of extracellular melanin was determined from the absorbance (475 nm) of (). The activation rate was calculated by the following formula. The same applies to the following examples. Activation rate (%) = 100 × (value of subject−value of control) /
Control value

The experimental results are shown in Table 1. As is clear from Table 1, the leaves and / or stems of pepper and betel showed a melanin production promoting action.

[0023]

[Table 1] Meanine amount is mean ± standard error, significant difference is *; p <0.
05, **; expressed as p <0.01.

Example 3 Tyrosinase activity promoting action The extract obtained in Example 1 was examined for tyrosinase activity promoting action.

Tyrosinase activity promotion test: DM
Dissolved in a 1: 9 solution of SO, pH 6.8 phosphate buffer,
This solution was diluted to adjust the concentration. Substrate Dopa solution (0.03%, dissolved in pH 6.8 phosphate buffer) 0.5
0.5 ml of the test solution so that the final concentration is 50, 200, 500 μg / ml (DMSO, p
0.5 ml of a 1: 9 solution of H6.8 phosphate buffer was used. ) Was added and incubated at 25 ° C. for 10 minutes.
0.5 ml of enzyme tyrosinase solution (30 U / ml, dissolved in pH 6.8 phosphate buffer) was added, and the absorbance after measuring for 5 minutes was measured at 475 nm to calculate the dopachrome production rate.

The experimental results are shown in Table 2. As is clear from Table 2, betel (leaves and stems) exhibited an action of promoting tyrosinase activity.

[0027]

[Table 2] Melanin amount is mean ± standard error, significant difference is **; p <
It is expressed as 0.01.

Example 4 Dopachrome's autoxidation promoting action in the melanin production process The dopachrome's autoxidation promoting action of the extract obtained in Example 1 was examined.

Autoxidation acceleration test: DMSO, p
It was dissolved in a 1: 9 solution of H6.8 phosphate buffer, and this solution was diluted to adjust the concentration. Substrate dopa solution (0.03
%, PH 6.8 dissolved in phosphate buffer) 0.5 ml 25
After incubating at 10 ° C. for 10 minutes, 0.5 ml of the enzyme tyrosinase solution (30 U / ml, dissolved in pH 6.8 phosphate buffer) was added and further incubated for 5 minutes. After that, the final concentration is 200, 500, 1000 μg / ml
Test solution 0.5 ml (as a control, DMS
0.5 ml of a 1: 9 solution of O, pH 6.8 phosphate buffer was used. ) Was added and incubated for 60 minutes. Then, stop the reaction with 0.2 ml of 1 mol / l hydrochloric acid,
A sedimentation was obtained by centrifugation at 3,000 rpm for 15 minutes. This precipitation is further performed once with 1 ml of 6 mol / l hydrochloric acid,
After washing with 2 ml of distilled water by centrifugation once, it was dissolved in 2 ml of solene (made by Packard), and when it was difficult to dissolve, it was forcibly dissolved with a sonicator. The absorbance of the solution was measured at 400 nm, and the amount of melanin was calculated from the absorbance curve of standard melanin.

The experimental results are shown in Table 3. As is clear from Table 3, the leaves and / or stems of pepper and betel showed an action of promoting autoxidation of dopachrome in the process of melanin production.

[0031]

[Table 3] Melanin amount is mean ± standard error, significant difference is **; p <
It is expressed as 0.01.

Example 5 Emollient Cream An emollient cream having the following composition was produced by a conventional method.

[0033] (Composition) (wt%) Stearic acid 2.0 Stearyl alcohol 7.0 Reduced lanolin 2.0 Squalene 5.0 Octyldecanol 6.0 Polyoxyethylene cetyl ether 3.0 Lipophilic type monooxystearate glycerin 2.0 Fragrance 0.3 Preservative Appropriate amount Antioxidant Appropriate amount Propylene glycol 5.0 Dry extract obtained in Example 1 1.0 Purified water amount of 100

Example 6 Emollient Lotion An emollient lotion having the composition shown below was produced by a conventional method.

[0035] (Composition) (wt%) Stearic acid 0.2 Cetanol 1.5 Vaseline 3.0 Lanolin alcohol 2.0 Liquid paraffin 10.0 Polyoxyethylene monooleate 2.0 Fragrance 0.3 Glycerin 3.0 Propylene glycol 5.0 Triethanolamine 1.0 Dry extract obtained in Example 1 1.0 Purified water amount of 100

[0036]

EFFECT OF THE INVENTION Pepper according to the present invention
The plants belonging to the family Ceae) and extracts thereof have an excellent melanin production promoting effect, and can prevent and / or treat melanin deficiency diseases such as leukoplakia and leukoplakia.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 43/00 111 A61P 43/00 111 F term (reference) 4C083 AA111 AC012 AC022 AC072 AC122 AC182 AC242 AC392 AC402 AC542 AD512 CC02 CC04 CC05 DD31 EE12 EE24 4C088 AB36 AC01 BA08 CA04 MA16 MA28 MA63 NA11 NA14 ZA89 ZB21 ZC02

Claims (7)

[Claims] 1. A melanin production promoter comprising a plant belonging to the family Piperaceae or an extract thereof. 2. A melanin production promoter containing a plant belonging to the genus Pipeer or the genus Peperomia or an extract thereof. 3. A tyrosinase activity promoter containing a plant belonging to the family Piperaceae or an extract thereof. 4. A tyrosinase activity promoter containing a plant belonging to the genus Pepper or the genus Peperomia or an extract thereof. 5. A preventive and / or therapeutic agent for melanogenesis deficiency, which comprises a plant belonging to the family Piperaceae or an extract thereof. 6. A preventive and / or therapeutic agent for melanogenesis, which comprises a plant belonging to the genus Pipeer or the genus Peperomia or an extract thereof. 7. The preventive and / or therapeutic agent for melanogenesis deficiency according to claim 5 or 6, wherein the dosage form is an external preparation for skin.