JP2002068933A - Fibroblast proliferation stimulator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new and effective fibroblast proliferation stimulator having high stability, and a skin care preparation and a bath medicine formulated with the same. SOLUTION: An extract of a plant of the genus Opuntia and/or a pressed juice of a plant of the genus Opuntia is found to have a high human-derived normal fibroblast proliferation stimulating action. Consequently, the extract and/or the pressed juice is applied as a fibroblast proliferation stimulator and is used also for a skin care preparation and a bath medicine.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

FIELD OF THE INVENTION The present invention relates to a novel and safe fibroblast growth promoter containing a prickly pear plant extract, and various skin external preparations and bath preparations in the pharmaceutical, quasi-drug or cosmetic fields. It is related to application to

[0002]

2. Description of the Related Art In aging skin, qualitative and quantitative changes occur in collagen fibers, elastin fibers, and acidic mucopolysaccharides, which are dermal matrix components, as fibroblast activity decreases. Collagen fibers become rigid due to the formation of abnormal aging crosslinks and lose their inherent elasticity. The elastin fiber denatures and disintegrates, and elastin having a different amino acid composition is produced compensatory and the dysfunction proceeds.
As a result, the skin loses its flexibility, causing wrinkles and sagging. In these physiologically aged skins, a decrease in proliferation ability and a decrease in physiological functions are observed, whereas in photoaged skin, fibroblasts increase and proliferate, and collagen production ability is also enhanced. One of the most distinctive features of this striking control is the thickness of the skin, where photoaged facial skin is strongly thickened,
Physiologically aged skin inside the forearm usually thins gradually. When considering the aging of human skin, it is necessary to use defense methods corresponding to these conflicting phenomena. However, enzymes are complexly functioning in vivo, and elucidation of the mechanism is an issue to be solved. To date, α-hydroxy acids, retinoic acids, fibroblast proliferating agents, female hormone-like substances, and the like have been developed and used for the purpose of preventing such skin aging. Chlorella water extract (JP-A-9-40523), sesame, sanyak,
Extracts include red pepper, red clover, pterodactyla, Bakumondou extract (JP-A-10-45615), almond, dandelion, Chinese elderberry, Sengoku, assembly, Souhakuhi, Tonin, carrot, hop, Mukuge, Yokuinin (JP-A-10-36279).

[0003]

However, the conventional fibroblast growth promoters have not yet yielded effective results when formulated into an actual product based on the concentration used and the growth rate. In view of these circumstances, the present inventors screened plant extracts and components that are useful as fibroblast proliferation promoters, and showed a high fibroblast proliferation promoting effect on prickly pear plant extracts and squeezed juice. The present inventors have further found that an antiaging effect can be expected from a skin external preparation and a bath containing the same and that they are safe in dermatology, and have completed the present invention.

[0004] Accordingly, an object of the present invention is to provide a highly safe external skin preparation and bath agent which has an excellent effect as a fibroblast proliferation promoter and is effective in preventing skin aging by blending these. To provide.

[0005]

The plant of the genus Prickly Pear used in the present invention is Cactaceae (Cactacea).
e) is classified into the prickly genus (Opuntia),
Ounta ficus-in
dica) (= Cactus fiction-indic
a) (= Cactus opuntia), Kimbusen (Opuntiatuna) (= Cactus pol)
yanthos), Tanshiniwa (Opuntia v)
ulgaris) (= Opuntia monacan
tha) (= Cactus urumbeba), (O
puntia streptacantha) can be cited as a typical example. Incidentally, it has not been known at all that the plant of the genus Cactus has a fibroblast proliferation promoting effect.

[0006] Other cactus genus, for example, Brassipuntia, Cephalovereus, Ceraceus, Epiphyta (Epiphy)
llum), genus Cactus (Harris)
ia), the genus Hylocereus, the genus Cactus (Melocactus), the genus Perilla, the genus Perescia, the genus Rhipsa (Rhipsa)
lis), a similar fibroblast proliferation promoting effect can be expected.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION A prickly pear extract used in the present invention refers to a stem, stem,
Leaves, flowers, fruits and roots may be used as needed, raw or dried as is or crushed, and the preparation method is not particularly limited. For example, various appropriate solvents may be used at room temperature or under heating. And the method of extraction.

Specific examples of the extraction solvent include water, lower monohydric alcohols such as methanol and ethanol, glycerin,
Liquid polyhydric alcohols such as propylene glycol and 1,3-butylene glycol; lower alkyl esters such as ethyl acetate; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; vegetable oils such as olive oil and jojoba oil; One or a mixture of two or more of these can be used. Among them, it is preferable to use water or a water-soluble solvent, in particular, one or a mixed solvent of water, ethanol, glycerin, propylene glycol, and 1,3-butylene glycol.

[0009] The compressed juice of Prickly Pear genus plant used in the present invention refers to the stem, stem,
A sap obtained by pressing leaves, flowers, fruits, and roots.

As the extract / pressed juice, the extract obtained as described above may be used as it is, or may be further concentrated or filtered as necessary. Also, extracts,
The pressed juice can be fractionated and purified by a conventional method, for example, a column chromatography method, a countercurrent distribution method or the like, and used.

Further, the extract / squeezed juice may be prepared by concentrating or freeze-drying the above extract, preparing a powder or paste, and formulating it as appropriate.

The prickly pear plant extract / squeezed juice of the present invention can be used as it is as a fibroblast growth promoting agent, and can also be incorporated into an external preparation for skin and a bathing agent. Absent. Although it depends on the type and quality of the product to be compounded and the degree of expected effect, it is preferably 0.0001 to 10.0% by weight (hereinafter simply referred to as “%”) in terms of dry solid content, particularly 0.001%. ~ 3.
0% is more preferable.

[0013] The fibroblast proliferation promoter, skin external preparation and bath preparation of the present invention are added to the essential components of the pear cactus plant extract / squeezed juice, and if necessary, in a range that does not impair the effects of the present invention. And pharmaceuticals, quasi-drugs, cosmetics, and other components used in pharmaceuticals and additives. Specific examples of these additional components are as follows.

As the surfactant, higher fatty acid soap,
Anionic surfactants such as alkyl sulfate salts, polyoxyethylene alkyl ether sulfate salts, acyl N-methyl taurine salts, alkyl ether phosphate ester salts, N-acyl amino acid salts, alkyl trimethyl ammonium chloride, dialkyl dimethyl ammonium chloride, chloride Cationic surfactants such as benzalkonium, amino alcohol fatty acid organic silicone resin, etc., amphoteric surfactants such as alkyl dimethyl amino acetate betaine, alkyl amide dimethyl amino acetate betaine, 2-alkyl-N-carboxy-N-hydroxyimidazolinium betaine Agent, polyoxyethylene alkyl ether, polyoxyethylene branched alkyl ether, sorbitan ester, polyoxyethylene sorbitan ester, glycerin fatty acid ester, polyoxy Ji Ren glycerol fatty acid ester,
Nonionic surfactants such as polyoxyethylene cholesterol ester, polyoxyethylene hydrogenated castor oil fatty acid ester, glycerin alkyl ether, polyoxyethylene glycerin alkyl ether, and polyglycerin fatty acid ester; natural interfaces such as lecithin, lanolin, cholesterol and saponin Activators and the like. As fats and oils,
Olive oil, camellia oil, macadamian nut oil, castor oil, sesame oil, safflower oil, peanut oil, vegetable oils such as pistachio oil, animal oils such as mink oil, egg yolk oil, jojoba oil,
Waxes such as carnaubaroe, candelilla wax, beeswax and lanolin, liquid paraffin, paraffin, petrolatum, hydrocarbons such as ceresin, microcrystalline wax, squalane, higher fatty acids such as lauric acid, myristic acid, stearic acid, isostearic acid, Higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, 2-octyldodecanol and jojoba alcohol; esters such as isopropyl myristate, 2-octyldodecyl myristate, cetyl 2-ethylhexanoate, and diisostearyl malate Kind,
Silicone oils such as methylpolysiloxane and methylphenylpolysiloxane; Polyhydric alcohols include ethylene glycol, polyethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin,
Polyglycerin, glucose, maltose, fructose, xylitol, sorbitol, maltotriose, erythritol and the like. Examples of the thickener include sodium alginate, xanthan gum, quince seed extract, guar gum, locust bean gum, sodium hyaluronate, collagen, casein, sodium carboxymethylcellulose, carboxyvinyl polymer and the like.

As the ultraviolet absorber, 2-hydroxy-
Benzophenone derivatives such as 4-methoxybenzophenone and 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, paraaminobenzoic acid derivatives such as paraaminobenzoic acid and octyl paradimethylaminobenzoate, octyl paramethoxycinnamate, mono-diparamethoxycinnamate
Methoxycinnamic acid derivatives such as glyceryl 2-ethylhexanoate, and salicylic acid derivatives such as homomenthyl salicylate. Preservatives include benzoate, salicylate, sorbate, dehydroacetate, paraoxybenzoate, benzalkonium chloride, hinokitiol, resorcinol, ethanol and the like. Examples of antioxidants include tocophenol, ascorbic acid, butylhydroxyanisole, dibutylhydroxytoluene, and gallic esters. Examples of the chelating agent include sodium ethylenediaminetetraacetate, sodium polyphosphate, and citric acid.

Furthermore, drugs having physiologically active effects such as antibacterial, cell activation, moisturizing, suppression of sebum secretion, inflammation, blood circulation promotion, astringency, antioxidation, whitening, etc., plant extracts, and extractable fractions and purified products thereof. They can be used together.

The agent system for the fibroblast proliferation promoter, skin external preparation and bath agent of the present invention may be any of a soluble system, an emulsion system, a powder dispersion system, a liquid dispersion system and the like.
Capsules, powders, granules, solids, liquids, gels, foams, sheets and the like may be used.

[0018] More specifically, medicinal preparations for internal and external use, basic cosmetics such as lotions, emulsions, creams and packs, shampoos, rinses, hair treatments, hair creams, pomades, hair sprays, hair dressings, hair dyes And cosmetics such as hair cosmetics, foundations, lipsticks, blushers, eye shadows, eye liners, bath agents, toothpastes, deodorants, hygiene articles, wet tissues, and the like.

[0019]

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

(Preparation Example 1) Prickly Pear Plant Opu
Purified water was added to 100 g of dried powder of the stem node of nia streptacantha, extracted at 80 ° C. for 5 hours, filtered with a filter aid, and filtered to obtain about 1 kg of extract A (2.4% solids). .

(Production Example 2) Opu cactus plant Opu
10 v / v% ethanol was added to 100 g of a freeze-dried powder of the fruit of nia streptacantha,
After extraction at 70 ° C. for 5 hours, filtration and concentration and drying, about 25 g of powder extract B is produced.

(Production Example 3) Prickly Pear Plant Opu
Purified water was added to 100 g of powder obtained by drying and pulverizing the fruit of N. ficus-indica, extracted at 70 ° C. for 8 hours, filtered, and about 1 kg of extract C (solid content: 3.1)
%).

(Production Example 4) Prickly Pear Plant Opu
Purified water was added to 1 kg of freshly prepared stem stalks of nia tuna in about 2 cm square, extracted at 70 ° C. for 8 hours, filtered with a filter aid, and extracted with about 1 kg of extract C (solid content 2%). .3%).

(Production Example 5) Prickly Pear Plant Opu
nia streptacantha raw stem 1k
g is squeezed and freeze-dried to obtain a powdered juice A2
Make 0 g.

(Test Example 1) Evaluation of the action of promoting the proliferation of fibroblasts
An evaluation test for promoting the proliferation of fibroblasts was carried out using the MTT reduction method. (TIM Mosmann; Journal
of Immunological Methods
p55-63, 1983)

(Test method) 10% FBS (fetal calf serum;
Human-derived normal skin fibroblasts (Clonetics) were seeded at a density of 3 × 10 ³ cells / well in a 96-well plate by using MEM (Gibco) containing MEM (purchased from Nikkei) at 37 ° C., 5% CO _2. For 72 hours. After removing the medium, the cells are washed with PBS (-) (Nissui Pharmaceutical), replaced with MEM containing 0.5% FBS to which each concentration of the test sample is added, and cultured at 37 ° C. and 5% CO _{2 for} 48 hours. At this time, the blank was 0.5 without the test sample.
% MEM containing FBS. After exchanging the medium every 48 hours and culturing for 2 days, 4 days, and 6 days, the absorbance difference at 570 nm and 620 nm was measured by the MTT reduction method. The cell growth rate of the added group was measured.

[0027]

[Table 1]

(Test Results) As shown in Table 1, the extract of Prickly Pear cactus and the pressed juice of Prickly Pear Cactus of the present invention were found to have a fibroblast proliferation promoting effect.

(Test Example 2) Panel Test A 50-year-old woman aged 30 to 55 years (average age: 40.5 years) was subjected to the following external composition containing a fibroblast growth promoter and a control external composition. The product was applied to the face twice a day (morning and evening) for three consecutive months and the sensory evaluation was shown in Table 2. Sensory evaluation was made on four items: moisture, texture, wrinkles, spots and freckles. As a control, the same amount of purified water as the fibroblast proliferation promoter of the present invention was used.

External composition for skin used in panel test 1.0% of fibroblast proliferation promoter of the present invention of Production Example 1 (or 0.1% of fibroblast proliferation promoter of the present invention of Production Example 5) glycerin 5. 0% Ethanol 5.0% Paraoxybenzoate 0.2% Purified water 84.8%

As is evident from Table 2, the composition for external use on skin of the present invention showed high efficacy, and the effect was derived from the fibroblast proliferation promoter of the present invention.

[0032]

[Table 2]

(Test Example 3) Acute toxicity test Sprague-Drawle of 5 males and 5 females fasted
y CD rats, males weighing 205-222 g, females 21
At 5 to 221 g, 8 to 12 weeks of age were used. The test sample is
The extract shown in Production Example 1 was used. Each rat was administered once by oral gavage at 2000 mg / kg based on the body weight after fasting at the time of administration. Rats were observed for death, or for obvious signs of toxicity, 30 minutes, 1 hour and 4 hours after administration, and once a day thereafter for 14 days. Body weight before administration on day 0, 7 days and 1
On day 4, individual recordings were made. At the end of the test, the rats were sacrificed by cervical dislocation and gross pathological examination was performed. No death or general toxicity symptoms were observed in the test results. Also, body weight by individual showed the expected weight gain in all rats throughout the study period. No abnormalities were observed at necropsy. By these, Sprague
-Acute oral 50% in DrawleyCD rats
The mortality (LD ₅₀ ) was confirmed to be greater than 2000 mg / kg body weight. Thus, it is clear that the fibroblast proliferation promoter of Production Example 1 is excellent in safety.

Test Example 4 Skin Safety Test The extract shown in Production Example 1 was used as a test solution. The primary skin irritation test was performed using three New Zealand White rabbits, and as a result, the primary skin irritation index was 0.0. The continuous skin irritation test was performed on white Du of 3 males and 3 females.
Using nkin Hartley guinea pigs, 14
As a result of repeated administration for consecutive days, the maximum weekly average stimulation index was 0.0, and it was determined to be non-stimulated. The eye irritation test was performed on the eyes of three New Zealand White rabbits, and the maximum group average score was 0.0, which was determined to be non-irritant.
The skin sensitization test was performed using Adju using white guinea pigs.
By the vant / Patch method, one test animal and one control animal were used.
The test was performed with 0 mice, and the expression rate was 0/10, and it was confirmed that the animals had no sensitization. Phototoxicity and photosensitization tests
When performed using five Dunkin Hartley guinea pig females each, it did not cause any signs of phototoxicity or photosensitization. From these skin safety results,
It is clear that the fibroblast proliferation promoter of Production Example 1 is extremely safe in terms of skin safety.

(Test Example 5) Patch test 30 men from 20 to 51 years old (average age 25.2 years)
A total of 60 healthy volunteers, each consisting of 30 women, were used. As the test substance, the extract of Preparation Example 1 and the external composition for skin used in the panel test were used, and physiological saline was used as a control. In the test, a 24-hour blockage application test was performed. Fin chamber for human body sticking test (diameter 11m)
m, Taisho Pharmaceutical Co., Ltd.).
Immediately after the application of L, it was stuck to the back of the subject and left for 24 hours. Then, the fin chamber was removed 24 hours later, and the skin reaction was observed 30 minutes later and the next day (48 hours later) according to the criteria.

The criterion was in accordance with the criterion of the Japanese Patch Test Study Group. No response ・ ・ ・ ・ ・ (−) Slight erythema ・ ・ ・ ・ ・ (±) Obvious erythema ・ ・ ・ ・ ・ ・ ・ (+) Erythema + swelling ・ ・ ・ ・ ・ (++) Erythema + swelling + papule or small Bubble ・ ・ ・ ・ ・ (++++) Large bubble ・ ・ ・ ・ ・ (++++)

Table 3 shows the test results. No example showing a positive reaction (+ or more) was found. In addition, the case where slight erythema (±) occurred was almost the same frequency as the case using physiological saline in each sample group. Therefore, the skin irritation was considered to be low, and it was clarified that there was no problem in terms of safety when blended into an external preparation for skin.

[0038]

[Table 3]

(Example 1) Cream The following components (1) to (10) and, separately, the following components (11) to (15) are heated and dissolved at 75 ° C. to obtain solution A and solution B, respectively. The solution A was added to the solution A, emulsified, cooled to 50 ° C. while stirring, and the component (16) was added to prepare a cream. (Components) (% by weight) (1) Jojoba oil 3.0% (2) Squalane 2.0% (3) Methylpolysiloxane 0.5% (4) Stearyl alcohol 0.5% (5) Cetyl alcohol 5% (6) glyceryl tri (capryl / caprate) 12.5% (7) glyceryl monostearate 5.0% (8) 1.5% diglyceryl monostearate (9) decaglyceryl monostearate 0% (10) Propyl parahydroxybenzoate 0.1% (11) Xanthan gum 0.1% (12) Fibroblast proliferation promoter of the present invention (Production Example 1) 1.0% (13) 1,3-butylene glycol 2.5% (14) Methyl paraoxybenzoate 0.2% (15) Purified water 67.5% (16) Fragrance 0.1%

Example 2 Lotion The following components (5) to (8) were mixed and dissolved to obtain a solution A. Separately, the following components (1) to (4) and (9) were mixed and dissolved. The solution (B) was used, and the solution A and the solution B were uniformly mixed to obtain a lotion. (Components) (% by weight) (1) Quince seed extract 8.0% (2) Glycerin 3.0% (3) 1,3-butylene glycol 5.0% (4) Fibroblast proliferation promoter of the present invention ( Production Example 4) 2.0% (5) Polyoxyethylene sorbitan laurate 1.2% (6) Ethyl alcohol 5.0% (7) Preservative 0.2% (8) Fragrance 0.1% (9 ) Purified water 75.5%

Example 3 Emulsion The following components (1) to (10), and separately from this, the following component (1)
Solution 1) to (14) and (16) were heated and dissolved at 75 ° C. to obtain solution A and solution B, respectively, and solution B was added to solution A, emulsified, and cooled to 50 ° C. with stirring to obtain component (15). Was added to prepare an emulsion. (Ingredients) (% by weight) (1) Jojoba oil 1.0% (2) Squalane 2.0% (3) Behenyl alcohol 1.0% (4) Tri (caprylic / capric acid) glyceryl 2.0% (5) Tetraglycerin condensed ricinoleic acid 0.1% (6) Propylene glycol monooleate 0.5% (7) Glyceryl monostearate 1.0% (8) Hexaglyceryl monomyristate 1.0% (9) Monomyristate Decaglyceryl 0.5% (10) Propyl parahydroxybenzoate 0.1% (11) Quince seed extract 5.0% (12) Fibroblast growth promoter of the present invention (Production Example 2) 0.1% (13) 1,3-butylene glycol 3.0% (14) Methyl paraoxybenzoate 0.1% (15) Fragrance 0.1% (16) Purified water 82.5%

(Example 4) Cleansing gel The following components (1) to (3), and separately from this, the following component (4)
(6) and (8) are heated and dissolved at 70 ° C. to obtain solution A and solution B, respectively, and solution B is added to solution A and stirred until uniform. While stirring, the mixture was cooled to 50 ° C., and the component (7) was added to prepare a cleansing gel. (Components) (% by weight) (1) Hexaglyceryl monomyristate 20.0% (2) Liquid paraffin 59.7% (3) Paraoxybenzoate 0.3% (4) Fibroblast proliferation promoter of the present invention (Production Example 3) 1.0% (5) Concentrated glycerin 5.0% (6) Sorbitol 5.0% (7) Flavor 0.1% (8) Purified water 8.9%

(Example 5) Packing agent The following components (1) to (4) and (9) are heated and dissolved at 70 ° C. to obtain a solution A, and (6) to (8) are mixed to obtain a solution B. B
After mixing the liquid with the liquid A, the mixture was cooled and (5) was uniformly dispersed to prepare a pack. (Components) (% by weight) (1) Polyvinyl alcohol 20.0% (2) Glycerin 5.0% (3) Kaolin 6.0% (4) Fibroblast proliferation promoter of the present invention (Production Example 1) 0% (5) Benzalkonium chloride 20.0% (6) Ethanol 20.0% (7) Methyl parahydroxybenzoate 0.2% (8) Fragrance 0.1% (9) Purified water 27.7%

Example 6 Shampoo The following components (1) to (5) and (13) were heated and mixed at 70 ° C. to make them uniform, and then (6) to (12) were added and mixed uniformly. A shampoo was prepared. (Ingredients) (% by weight) (1) N-coconut oil fatty acid glutamate triethanol 25.0% (2) lauric acid diethanolamide 5.0% (3) potassium myristate 5.0% (4) ethylene glycol distearate 2.0% (5) Polyethylene glycol 400 15.0% (6) Silicon oil 1.0% (7) Fibroblast growth promoter of the present invention (Production Example 1) 1.0% (8) Chlorxylenol 1% (9) Vitamin E 0.1% (10) Olive leaf extract 2.0% (11) Methyl parahydroxybenzoate 0.2% (12) Fragrance 0.3% (13) Purified water 43.3%

Example 6 Bath Agent The following components were uniformly mixed to prepare a bath agent. (Ingredient) (% by weight) (1) Dry sodium sulfate 40.0% (2) Sodium bicarbonate 57.3% (3) Fragrance 2.0% (4) Light anhydrous silicic acid 0.3% (5) bottles Invention fibroblast proliferation promoter (Production Example 2) 0.2% (6) Yellow No. 202 (1) 0.2%

[0046]

EFFECTS OF THE INVENTION It is clear that the extract of prickly pear and / or the compressed juice of prickly pear of the present invention has a high human normal fibroblast proliferation promoting action, and the results of various safety tests show High safety was recognized. In addition, the external preparation for skin formulated from the panel test is effective in keeping the skin moist and preventing rough skin, keeping the texture of the skin, preventing wrinkles and preventing skin aging, and it is effective for all forms of skin external preparation Is applicable.

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Claims (4)

[Claims] An agent for promoting fibroblast proliferation, which comprises a prickly pear plant extract or / and a prickly pear plant press. 2. The plant of the genus Prickly pear is Opuntia.
The agent for promoting fibroblast proliferation according to claim 1, which is streptacantha Lemaire. 3. An external preparation for skin and a bath preparation comprising the fibroblast proliferation promoter according to claim 1. 4. An external preparation for skin and a bath preparation, comprising the fibroblast proliferation promoter according to claim 2.