JP6026717B2 - Hyaluronic acid production promoter - Google Patents

Description

The present invention relates to a hyaluronic acid production promoter, and more specifically, by promoting hyaluronic acid production of human skin fibroblasts, aging delay or prevention of skin, abnormal degradation of hyaluronic acid, or a decrease in hyaluronic acid production ability The present invention relates to a hyaluronic acid production promoter that can be used for treatment and is safe for a living body.

Hyaluronic acid is a constituent component of connective tissues such as animal skin, blood vessel wall, eye vitreous, and umbilical cord, and is also a main component of joint fluid. Hyaluronic acid has a function to retain moisture in the cell gap, a function to retain cells by forming a jelly-like matrix in connective tissue, a function to maintain the lubricity and flexibility of connective tissue, and external forces such as mechanical failure It has many functions such as a resistance function.

However, hyaluronic acid in dermal fibroblasts decreases with age, resulting in aging phenomena such as fine wrinkles and rustling. In order to improve such aging skin, many cosmetics containing a moisturizing agent such as hyaluronic acid have been proposed. For example, an osmotic pressure of 200 to 600 milliosmoles containing a moisturizing agent such as 1,3-butylene glycol, polyethylene glycol, maltitol, sorbitol, fructose, glucose, polyoxyethylene methyl glucoside, hyaluronic acid, etc. There are cosmetics (see, for example, Patent Document 1). However, these cosmetics only provide a moisturizing effect on the skin surface, and do not essentially improve or prevent aging skin. In addition, vitamins and herbal medicines are used as skin cell activators to improve aging skin. For example, a fibroblast growth promoter (for example, see Patent Document 2) containing a plant extract such as sesame, sanjak, capsicum, toki, dokudami, bakmondo, etc., and a metabolism promoter (for example, patent document) containing collagen protein. 3). However, none of them have been used to treat aged skin.

Further, hyaluronic acid in the joint fluid covers the surface of the articular cartilage and is useful for smooth operation of the joint function. The amount of hyaluronic acid in fetal joint fluid is the largest, containing about 1 to 3 mg / ml, but the amount of hyaluronic acid in adult joint fluid is reduced to about 0.3 mg / ml. In the case of rheumatoid arthritis, the amount of hyaluronic acid in the joint fluid is further reduced, and at the same time, the viscosity of the joint fluid is significantly reduced. It is also known that the amount of hyaluronic acid in joint fluid decreases in purulent arthritis and gouty arthritis.

In these diseases, it is conceivable to increase the amount of hyaluronic acid in the joint fluid in order to improve the lubrication function, cover and protect the articular cartilage, suppress pain, and improve the properties of the pathological joint fluid. For example, when joint injection therapy with sodium hyaluronate is performed on patients with rheumatoid arthritis, traumatic arthritis, osteoarthritis and osteoarthritis, improvement of the above symptoms is observed (see, for example, Non-Patent Document 1). ) Has been reported. However, treatment of the above diseases is long-lasting and requires a doctor's prescription. Therefore, a drug containing a hyaluronic acid production promoter that can be easily treated in daily life has been desired.

In the healing process after burn injury, hyaluronic acid is known to increase significantly in the granulation during the period from the beginning of the granulation tissue growing from below the necrotic tissue until the entire tissue is replaced with granulation tissue. Hyaluronic acid production promoters are also expected as an early treatment for burns. In addition, it has been reported that hyaluronidase activity, which is a hyaluronic acid-degrading enzyme, is remarkably increased at sites such as gingivitis and hepatitis (for example, see Non-Patent Document 2). According to this, a decrease in the amount of hyaluronic acid due to an increase in hyaluronidase activity is considered to worsen the symptoms of gingivitis.

Various hyaluronic acid synthesis promoting substances have been found to improve the state caused by the reduction of hyaluronic acid. For example, hyaluronic acid production promoters containing compounds having isoprenoid chains such as β-carotene and tocopherol as essential components (for example, see Patent Document 4), hyaluronic acid production promoters containing conagenin as active ingredients (for example, patent documents) 5), an aloe extract, an okra extract, a water-soluble β-1,3-glucan derivative, a yeast extract and the like, a hyaluronic acid synthesis promoter (see, for example, Patent Document 6), Biohyaluronic acid synthesis promoter containing an extract of seaweed belonging to (for example, see Patent Document 7), hyaluronic acid synthesis promoter containing a lavender extract (see, for example, Patent Document 8), Davilia family Davilia genus Known is a biological hyaluronic acid synthesis promoter (see, for example, Patent Document 9) containing an extract of seaweed belonging to Yes.

As other components having hyaluronic acid production promoting action, for example, combined use of retinol and a thermolysin degradation product of soybean protein (see, for example, Patent Document 10), protease hydrolysis product of soybean protein (see, for example, Patent Document 11). ), A combination of retinols and specific peptides having a specific amino acid sequence (see, for example, Patent Document 12), an extract of seaweed Anaaaosa (for example, see Patent Document 13), a plant extract of Mesoy (for example, Patent Document 14), extracts of seaweeds belonging to the genus Funori (for example, refer to Patent Document 15), Aosa family (Ulva) (excluding Ulva pertusa), Aosa family Aonori (Enteromorpha), Ganodermaceae (Gracilaria), Ten Gelidium (Gelidium), Milinaceae (Eucheuma), Compositae (Eisenia), Ainuweidae (Undaria), Honda (Hondaki) or Ascophyll (Ascophylla) Seaweed extract (see, for example, Patent Document 16), N-methyl-L-serine, ethanolamine, N-methylethanolamine (see, for example, Patent Document 17), mammal (human, cow, goat, sheep) And porcine whey processed under specific conditions (see, for example, Patent Document 18), collagen or gelatin degradation product of collagenase, and consisting of a collagen tripeptide represented by a specific amino acid sequence Certain collagen peptides (eg Patent Document 19), extracts of acidic xylo-oligosaccharides having a uronic acid residue in the xylo-oligosaccharide molecule (see, for example, Patent Document 20), Changbut, Marbatichichanoki, Ryukyu-chanoki and Tishanoki (see, for example, Patents) Document 21), specific phosphate derivatives (for example, see Patent Document 22), and the like are known.

On the other hand, chondroitin sulfate is usually marketed as a sodium salt as a moisturizer, feel improver, thickener, emulsion stabilizer and the like, and is used in cosmetics, quasi drugs, pharmaceuticals, foods and the like. As such, oil-in-water emulsion lotions for moisturizing the skin (see, for example, Patent Document 23) and diseases or conditions associated with collagen fibril formation such as wounds (ulcers) are treated. For example (see, for example, Patent Document 24) and elasto-viscous solutions for ophthalmic surgery that reduce the risk of increased intraocular pressure (see, for example, Patent Document 25). In addition, it is known that normal skin fibroblasts derived from connective tissue have a hyaluronic acid degrading activity, and the degradation is promoted by a chondroitin sulfate C derivative (see, for example, Patent Document 26). However, it has not been known that chondroitin sulfate itself has an action of promoting hyaluronic acid production by human skin fibroblasts.

Japanese Patent Laid-Open No. 5-170641 Japanese Patent Laid-Open No. 10-45615 Japanese Patent Laid-Open No. 7-278012 JP 2004-359573 A International Publication No. 2007/11066 JP 2004-51533 A JP 2000-136147 A JP-A-10-182402 Japanese Patent Laid-Open No. 09-176036 JP 2008-24704 A International Publication No. 2007/49400 International Publication No. 2007/148804 JP-A-6-9422 JP-A-9-87163 JP-A-8-198741 JP 7-101871 A JP-A-6-189780 JP 11-335234 A Japanese Patent Laid-Open No. 2004-123637 JP 2004-59480 A JP 2002-363087 A JP 2002-363081 A JP-A-11-180821 Special table 2007-534749 gazette Japanese Patent No. 3224815 Japanese Patent Laid-Open No. 11-80205

Okuyama Takashi et al., “Animal Component Utilization Collection, Terrestrial Animals” R & D Planning Co., Ltd., June 25, 2002, p753-755 Kakegawa Toshio et al., “Inflammation”, Volume 4 (1984) No. 4, p437-438

The object of the present invention is to promote hyaluronic acid production of human skin fibroblasts in daily life, thereby delaying or preventing skin aging, abnormal degradation of hyaluronic acid, or a decrease in hyaluronic acid production ability, for example, An object of the present invention is to provide a hyaluronic acid production promoter that can be used for the treatment of rheumatism, osteoarthritis, gingivitis and the like and is safe for the living body.

As a result of intensive studies in view of the above circumstances, the present inventors have found that chondroitin sulfate has a hyaluronic acid production promoting action, and completed the present invention. That is, this invention is a hyaluronic acid production promoter mix | blended with cosmetics with N-acetylglucosamine, Comprising: It is a hyaluronic acid production promoter of the human skin fibroblast which contains chondroitin sulfate as an active ingredient .

Since the hyaluronic acid production promoter of the present invention can promote hyaluronic acid production of human skin fibroblasts in daily life, it is a disease caused by delayed or preventing skin aging, abnormal degradation of hyaluronic acid, or a decrease in the ability to synthesize hyaluronic acid For example, it can be applied to the treatment of rheumatism, osteoarthritis, gingivitis and the like, and is safe for the living body.

It is a graph which shows the hyaluronic acid production ability of this invention. It is a graph which shows skin elasticity.

Hereinafter, the present invention will be described in detail.

Examples of chondroitin sulfate used in the hyaluronic acid production promoter of the present invention include chondroitin sulfate A, chondroitin sulfate B, chondroitin sulfate C, chondroitin sulfate D, chondroitin sulfate E, chondroitin sulfate K, and salts thereof. Examples of the chondroitin sulfate salt include alkali metal salts such as lithium salt, potassium salt, sodium salt and rubidium salt, and alkaline earth metal salts such as beryllium salt, magnesium salt, calcium salt and strontium salt. The chondroitin sulfate is obtained from animals such as sharks, sharks, squids, whales, cows and pigs, and usually parts such as cartilage and skin are used. A commercial item can be used for the chondroitin sulfate. For example, “chondroitin sulfate A. sodium salt (whale cartilage) .SSG”, “chondroitin sulfate A. sodium salt (whale cartilage) .SG”, “chondroitin sulfate A. sodium salt (sturgeon chord)”, “chondroitin sulfate B. Sodium salt (pig skin), “chondroitin sulfate C. sodium salt (shark cartilage). SSG”, “chondroitin sulfate C. sodium salt (shark cartilage). SG”, “chondroitin sulfate D sodium salt (shark cartilage)”, “Chondroitin sulfate E. sodium salt (squid cartilage)” and the like (above, Biochemical Biobusiness Co., Ltd.), chondroitin sulfate sodium derived from Aki nose nasal cartilage and mixed with chondroitin sulfate A and chondroitin sulfate C “MC marine chondroitin And “SCS Marine Chondroitin” (Both Japan Barrier Free Co., Ltd.), “Sodium Chondroitin Sulfate (Outer Base)”, “Sodium Chondroitin Sulfate (Outside Base) FDT-P” and “Sodium Chondroitin Sulfate (Outside Base) FDC-” P "(above, Biofac A / S) and the like. Among the chondroitin sulfates, chondroitin sulfate A, chondroitin sulfate C, and sodium salts thereof are preferable, but the mechanism of action is not clear at present. These chondroitin sulfates are blended in the hyaluronic acid production promoter of the present invention with one kind or two or more kinds as active ingredients. Chondroitin sulfate is a substance that has been used in cosmetics, quasi-drugs, pharmaceuticals, foods, and the like, has no safety problems, and is excellent in stability.

The chondroitin sulfate can be obtained, for example, by the following production method. First, cartilage or skin derived from salmon, shark, squid, whale, cow or pig is appropriately used as a raw material. Next, this raw material is subjected to a conventional enzyme treatment to remove proteins. Furthermore, a decalcification process, a degreasing process, a decoloring process, and a deodorizing process by a conventional method are performed. Trichloroacetic acid is added to the solution obtained through each treatment step, and the final concentration of trichloroacetic acid is adjusted to 10%. The prepared solution is allowed to stand at 4 ° C. for 24 hours and then centrifuged, and the resulting supernatant is dialyzed for 24 hours. 10% sodium acetate is added to the dialyzed internal solution, ethanol is added to the resulting supernatant, and the resulting precipitate is recovered as a crude extract of chondroitin sulfate. The crude extract of chondroitin sulfate is purified using a gel filtration chromatography support “Sephadex G-25” (GE Healthcare Japan, Inc.), and the resulting purified product is recovered as a powder.

As the dosage form of the hyaluronic acid production promoter of the present invention, in addition to the powder obtained by the above production method example, it can be made into a liquid using an appropriate solvent or made into a granule using an appropriate excipient. What is formulated into a state that is easy to be used may be used.

In addition to the above chondroitin sulfate, which is an active ingredient, the hyaluronic acid production promoter of the present invention includes components that are usually used in cosmetics or pharmaceuticals or components that are allowed to be used depending on the purpose, and that are effective for the present invention. It can mix | blend suitably within the range which is not impaired.

In cosmetics or pharmaceuticals containing the hyaluronic acid production promoter of the present invention, the compounding amount of chondroitin sulfate as an active ingredient is 0.005% by mass (hereinafter referred to as wt) in terms of dry solids based on the total amount of cosmetics or pharmaceuticals. %) And 10 wt% or less are preferable from the viewpoint of the effect and cost. In particular, 0.005 wt% or more and 5 wt% or less is preferable when blended and applied to a product.

When the hyaluronic acid production promoter of the present invention is blended with cosmetics or pharmaceuticals, as other components to be blended with the hyaluronic acid production promoter, all can be used as long as they can be normally used in cosmetics or pharmaceuticals, respectively. It chooses suitably according to.

The form of the cosmetic product of the present invention is not particularly limited, but basic cosmetics such as creams, emulsions, lotions, essences, facial cleansing agents, cleansing materials, packs, makeup cosmetics such as lipsticks, foundations, and eye colors, It can be used as toiletries such as body soaps, soaps, shampoos, rinses, conditioners, and hair cosmetics such as hair setting agents. Since the cosmetic containing the hyaluronic acid production promoter of the present invention is excellent in the effect of preventing skin aging, a cosmetic for external use is preferred.

Examples of the cosmetics include excipients and fragrances generally used in cosmetics, fats and oils, surfactants, moisturizers, whitening agents, pH adjusters, binders, polyhydric alcohols, essential oils. And various cosmetic ingredients such as fragrances, thickeners, preservatives, antioxidants, ultraviolet absorbers, pigments, plant pulverized products and herbal medicines, inorganic salts and inorganic acids, cleaning agents, emulsifiers, and the like. .

The cosmetic product of the present invention is preferably formulated with N-acetylglucosamine among hygienic acid production promoters containing chondroitin sulfate as an active ingredient among the cosmetic components that can be blended. When both are used in combination, the effect of improving aged skin such as skin elasticity becomes more excellent. The blending amount of N-acetylglucosamine is not particularly limited, but is preferably 0.005 wt% or more and 10 wt% or less in terms of dry solid content based on the total amount of cosmetics in view of the effect and cost.

The dosage form of the pharmaceutical product of the present invention is not particularly limited, and it may be an oral administration formulation or a parenteral administration formulation. Specifically, aerosols, liquids, extracts, elixirs, capsules (hard capsules, soft capsules, microcapsules), granules, pills, ophthalmic ointments, transdermal preparations, suspensions, emulsions, seats Preparation (vaginal preparation), powder, spirits, tablets (plain tablet, coated tablet, special tablet), syrup, soaking agent, decoction, injection (water-soluble injection, water-insoluble injection), patch, tincture Agents, eye drops, lozenges, ointments, poultices, fragrances, liniments, limonades, flow extracts, lotions and the like. These preparations can be produced by a conventional method in the pharmaceutical technical field, for example, by the method described in the Japanese Pharmacopoeia. These preparations can be safely administered to mammals including humans.

The pharmacologically acceptable carrier is not particularly limited, and various carrier substances known as pharmaceutical materials can be used. The carrier substance is not particularly limited, and examples of solid carriers include excipients, lubricants, binders, and disintegrants. In addition to the carrier substance, formulation additives such as preservatives, colorants, natural pigments, sweeteners and the like can be used as necessary. These substances include lactose, hydroxypropylmethylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, ethylcellulose, corn starch, crystalline cellulose, carmellose calcium, anhydrous silicic acid, synthetic aluminum silicate, magnesium stearate, talc, etc. Although it can illustrate concretely, it is not restricted to these.

The carrier substance in the liquid preparation is not particularly limited, and examples thereof include a solvent, a solubilizing agent, a suspending agent, a tonicity agent, a buffering agent, and the like. Furthermore, formulation additives such as preservatives, colorants, water-insoluble lake pigments, sweeteners and the like can be used as necessary. These substances include isotonic agents such as mannitol, sodium chloride, glucose, sorbitol, glycerol, xylitol, fructose, maltose, mannose, stabilizers such as sodium sulfite, preservatives such as benzyl alcohol and methyl parahydroxybenzoate. In addition to the above, a solubilizer, a soothing agent, a pH adjuster and the like can be specifically exemplified, but are not limited thereto.

Hereinafter, the present invention will be specifically described by illustrating examples and comparative examples. In addition, this invention is not limited at all by these Examples.

(Extraction and preparation of chondroitin sulfate)
The salmon nasal cartilage was subjected to an enzyme treatment by a conventional method to remove proteins. Next, the decalcification process, the degreasing process, the decoloring process, and the deodorizing process by the conventional method were performed. Trichloroacetic acid was added to the solution obtained through each treatment step to adjust the final concentration of trichloroacetic acid to 10%. The prepared solution was allowed to stand at 4 ° C. for 24 hours and then centrifuged, and the resulting supernatant was dialyzed for 24 hours. 10% sodium acetate was added to the dialyzed internal solution, ethanol was added to the resulting supernatant, and the resulting precipitate was recovered as a crude extract of chondroitin sulfate. The crude extract of chondroitin sulfate was purified using a gel filtration chromatography carrier “Sephadex G-25” (GE Healthcare Japan, Inc.), and the resulting purified product was recovered as a powder. The collected chondroitin sulfate was used for the following evaluation tests.

(Evaluation Test 1: Measurement of hyaluronic acid production ability)
The number of normal human fibroblasts (Kurashiki Boseki Co., Ltd.) was adjusted to 1 × 10 ⁵ cells / ml in a MEM medium containing 10% non-immobilized FBS. The prepared cells were seeded at 0.5 ml each in a 24-well plate (Nippon Becton Dickinson Co., Ltd.) and statically cultured at 37 ° C. in a 5% CO ₂ gas phase for 48 hours.

Chondroitin recovered in powder form in a MEM medium containing 0.25% non-immobilized FBS so that the final concentration of chondroitin sulfate is 0.2 mg / ml, 0.4 mg / ml, and 0.8 mg / ml. Sulfuric acid was prepared. The culture supernatant of the well plate that was allowed to stand for 48 hours was removed by aspiration, and 0.5 ml of each prepared chondroitin sulfate was added to each well (4 wells at each concentration). Wells to which chondroitin sulfate was not added were used as controls (4 wells). After standing culture for 72 hours, the culture supernatant was collected, and the hyaluronic acid content in the culture supernatant was measured with a commercially available hyaluronic acid measurement kit (Seikagaku Corporation).

The obtained data was subjected to data significance test using t-test. * (P <0.05) is shown at 5% significance level. The results are shown in FIG. In addition, the amount of hyaluronic acid at each concentration is a relative value when the amount of hyaluronic acid in the control is 100%.

As is apparent from the results shown in FIG. 1, chondroitin sulfate promoted hyaluronic acid production in a concentration-dependent manner.

(Evaluation test 2: mRNA measurement of hyaluronic acid synthase)
The number of normal human fibroblasts (Kurashiki Boseki Co., Ltd.) was adjusted to 1 × 10 ⁵ cells / ml in a MEM medium containing 10% non-immobilized FBS. 2 ml of the prepared cells were seeded in three 6-well plates (Nippon Becton Dickinson Co., Ltd.), and statically cultured at 37 ° C. in a 5% CO ₂ gas phase for 48 hours.

The final concentration of chondroitin sulfate is 0.1 mg / ml, 0.2 mg / ml, 0.4 mg / ml, and 0.8 mg / ml in MEM medium containing 0.25% non-immobilized FBS. Then, chondroitin sulfate recovered as a powder was prepared. The culture supernatant of the well plate that had been allowed to stand for 48 hours was removed by aspiration, and 2 ml of each adjusted chondroitin sulfate was added to each well (3 wells at each concentration). Wells to which chondroitin sulfate was not added were used as controls (3 wells).

After further stationary culture for 72 hours, the culture supernatant of the well plate was removed by aspiration, and 0.5 ml of RNA stabilization solution (Life Technologies Inc.) was added to each well. The cell solution was taken from each well with a scraper, and the total RNA was extracted with a commercially available total RNA purification kit (Qiagen “RNeasy Mini Kit”). At the time of extraction, the RNA amount of each cell solution was measured with a spectrophotometer, and the total RNA was adjusted so that the total RNA concentration was 200 μg / ml.

Using this total RNA as a template, the mRNA expression levels of GAPDH and HAS2 (Hyaluronan Synthase 2) as internal standards were measured. Detection is performed using a commercially available RT-PCR reagent kit (Takara Bio Inc. “SYBR Prime Script RT-PCR kit II (Perfect Real Time)”) using a commercially available real-time PCR apparatus (Cepheid “Smart Cycler”). The RT-PCR reaction was performed. The expression level of HAS2 is calculated based on the total RNA value, and the correction value is calculated with the value of GAPDH. Further, when the correction value without addition of the sample is set to 100, the expression level (%) of HAS2 mRNA expression by chondroitin sulfate addition is calculated. Calculated.

HAS2 mRNA expression promotion rate (%) = A / B × 100
A: Correction value when chondroitin sulfate is added B: Represents the correction value when no sample is added.

As is clear from Table 1, the addition of chondroitin sulfate was confirmed to promote the expression level of HAS2 mRNA. Thus, it was found that chondroitin sulfate acts on the gene of hyaluronic acid synthase HAS2 and promotes hyaluronic acid.

Examples of cosmetics and pharmaceuticals containing the hyaluronic acid accelerator of the present invention are shown below. The composition is expressed in wt%. In the following examples, “SCS marine chondroitin” (Nihon Barrier Free Co., Ltd.) was used as sodium chondroitin sulfate.

(Example 1) Toner lotion wt%
Sodium chondroitin sulfate 0.1
N-acetylglucosamine 0.05
Ethanol 7.5
Placenta extract 0.05
D-pantothenyl alcohol 0.1
Dipotassium glycyrrhizinate 0.1
Phenoxyethanol 0.3
Polyoxyethylene sorbitan monolaurate (20E.O.) 0.3
Polyoxyethylene hydrogenated castor oil 0.05
Edetate disodium 0.05
Concentrated glycerin 3.5
1,3-butylene glycol 2.5
Polyglycerin 0.5
Xanthan gum 0.5
Sorbit liquid 5.0
Carboxyvinyl polymer 0.1
Purified water to 100

(Example 2) Latex wt%
Sodium chondroitin sulfate 0.1
D-pantothenyl alcohol 0.1
Dipotassium glycyrrhizinate 0.1
Liquid paraffin 10.0
Concentrated glycerin 5.0
Dipropylene glycol 10.0
Sorbit liquid 5.0
White beeswax 0.5
Liquid paraffin 1.5
Lipophilic glyceryl monostearate 0.1
Stearic acid 0.3
Methyl polysiloxane 1.0
Vaseline 1.0
Cholesterol 0.02
Bentonite 0.1
N-stearoyl-L-glutamate sodium 0.3
Xanthan gum 0.2
Behenyl alcohol 0.3
Phenoxyethanol 0.2
Sodium hyaluronate (2) 0.05
Water-soluble collagen (F) 0.1
Vaginal ovary extract 0.1
Paraoxybenzoic acid ester 0.1
N-acetylglucosamine 0.05
Purified water to 100

(Example 3) Cream wt%
Sodium chondroitin sulfate 0.1
D-pantothenyl alcohol 1.0
Dipotassium glycyrrhizinate 0.5
Liquid paraffin 4.0
White beeswax 0.5
Cholesterol 1.0
Methylcyclopolysiloxane 2.0
Dipropylene glycol 7.0
Lipophilic glyceryl monostearate 0.3
Sodium cetyl sulfate 1.0
Polyethylene glycol monostearate 1.0
Sodium hyaluronate (2) 0.5
Water-soluble collagen (F) 0.1
Phenoxyethanol 0.5
Edetate disodium 0.05
N-acetylglucosamine 0.05
Purified water to 100

(Example 4) Nutritional cream wt%
Sodium chondroitin sulfate 1.0
Stearic acid 2.5
Stearyl alcohol 8.0
Squalane 5.0
Octyldodecanol 7.0
Glyceryl monostearate 4.0
Preservative Appropriate amount of perfume Appropriate amount of glycerin 6.0
Sorbitol 13.0
Potassium lactate 0.5
Purified water to 100

(Example 5) Lip treatment wt%
Sodium chondroitin sulfate 0.1
Candelilla Row 12.0
Solid paraffin 8.0
Beeswax 5.0
Carnavalo 2.0
Lanolin 14.0
Mannit 15.0
Isopropyl myristate 10.0
Perfume proper amount antioxidant proper amount castor oil to 100

(Example 6) Latex wt%
Sodium chondroitin sulfate 0.05
D-pantothenyl alcohol 0.1
Dipotassium glycyrrhizinate 0.1
Concentrated glycerin 5.0
Dipropylene glycol 10.0
Sorbit liquid 5.0
White beeswax 0.5
Liquid paraffin 1.5
Lipophilic glyceryl monostearate 0.1
Stearic acid 0.3
Methyl polysiloxane 1.0
Vaseline 1.0
Cholesterol 0.02
Bentonite 0.1
N-stearoyl-L-glutamate sodium 0.3
Xanthan gum 0.2
Behenyl alcohol 0.3
Phenoxyethanol 0.2
Sodium hyaluronate (2) 0.05
Water-soluble collagen (F) 0.1
Vaginal ovary extract 0.1
Paraoxybenzoic acid ester 0.1
N-acetylglucosamine 0.04
Purified water to 100

(Example 7) Pack wt%
Sodium chondroitin sulfate 0.5
Talc 25.0
Propylene glycol 6.0
Potassium chloride 0.1
Perfume proper amount kaolin to 100

(Example 8) Nutritional cream wt%
Sodium chondroitin sulfate 1.0
Stearic acid 2.0
Stearyl alcohol 8.0
Reduced lanolin 2.0
Squalane 8.0
Octyldodecanol 6.0
Preservatives Appropriate perfumes Appropriate propylene glycol
Glycerin 3.0
Lactic acid 0.3
Sodium citrate 0.5
Purified water to 100

(Example 9) Slimming Massage Gel wt%
Sodium chondroitin sulfate 2.0
Glycerin 60.0
Polyethylene glycol 200 14.0
Polyvinyl alcohol 0.2
Purified water to 100

(Example 10) Slimming massage soap wt%
Chondroitin sulfate sodium 5.0
Lauric acid 10.0
Potassium hydroxide 2.8
Coconut oil fatty acid potassium 30.0
Edetate disodium 0.15
Methylcellulose 0.5
Aminopropyl betaine laurate solution 2.0
Glycerin 8.0
Perfume proper amount purified water to 100

(Example 11) Massage gel wt%
Sodium chondroitin sulfate 0.5
Caffeine 0.1
Carboxyvinyl polymer 0.5
Polyvinyl alcohol 3.0
Glycerin 35.0
Polyethylene glycol 400 45.0
Ethanol appropriate amount Preservative appropriate amount purified water to 100

The production methods of Examples 1 to 11 were in accordance with ordinary cosmetic methods. Both cosmetics have improved skin wrinkles, moisture content, elasticity, and so on.

(Example 12) Granules wt%
Sodium chondroitin sulfate 10.0
Starch 30.0
Lactose 49.0
Crystalline cellulose to 100

The above respective parts by weight were uniformly mixed and prepared by a conventional method to prepare granules. As sodium chondroitin sulfate, “sodium chondroitin sulfate (external group) FDT-P” (BiofacA / S) was used. When 2 g of 1 capsule (200 mg of chondroitin sulfate sodium salt) was taken 3 times a day after meals to osteoarthritic patients, osteoarthritic pain was relieved after 6 months.

(Example 13) Tablet wt%
Sodium chondroitin sulfate 10.0
Crystalline cellulose 1.5
Vitamin C 20.0
Fragrance 1.0
Guam gum 0.1
Sucrose fatty acid ester 1.5
Powdered sugar to 100

Each said weight part was mixed uniformly, it produced by the conventional method, and was set as the tablet. As sodium chondroitin sulfate, “sodium chondroitin sulfate (external group) FDT-P” (BiofacA / S) was used. When 1 tablet 1500 mg (150 mg chondroitin sulfate sodium) was taken every night before going to bed, pain in the bones and joints was relieved after 6 months.

(Example 14) Soft capsule wt%
Sodium chondroitin sulfate 20.0
Vitamin E 20.0
Wheat germ oil 15.0
Glycerin fatty acid ester 5.0
Beeswax 2.0
Perilla oil to 100

The above ingredients were mixed and filled into a ceratin film made of gelatin and glycerin to obtain a soft capsule according to a conventional method. In addition, "sodium chondroitin sulfate (external group) FDC-P" (BiofacA / S) was used as sodium chondroitin sulfate. When 1 capsule 500 mg (100 mg of chondroitin sulfate sodium salt) was taken after meals in the morning and evening, the pain in the bone and joints was relieved after 6 months.

(Example 15) Hard capsule wt%
Chondroitin sodium sulfate 15.0
Sodium ascorbate 18.0
Dextrin 24.0
Glycerin fatty acid ester 0.5
Powdered sugar to 100

The above components were mixed and filled into a capsule container made of gelatin to obtain a hard capsule according to a conventional method. When 1 capsule 500 mg (sodium chondroitin sulfate 75 mg) was taken every night before going to bed, pain in patients with rheumatoid arthritis was relieved after 6 months.

(Comparative Examples 1-3)
The lotions, emulsions and creams of Comparative Examples 1 to 3 were obtained in the same manner as in Examples 1 to 3 except that sodium chondroitin sulfate and N-acetylglucosamine were not blended.

(Practical test: skin elasticity)
It divided into the P group which uses the lotion, milky lotion, and cream of Examples 1-3, and the Q group which uses the cosmetics, milky lotion, and cream of Comparative Examples 1-3. For the P group and Q group, 18 men, 10 men and 8 women, continued for 6 weeks. The elasticity of the skin before and after continuous use was measured using a skin viscoelasticity measuring apparatus (Courage + Khazaka Electronic GmbH “Cumeter MPA580”).

The obtained data was subjected to data significance test using t-test. * (P <0.05) is shown at 5% significance level. The results are shown in FIG. In addition, the value of each group is a relative value when the value before continuous use is set to 100.

As is clear from the results shown in FIG. 2, the skin elasticity of both the P group and the Q group was improved by continuous use for 6 weeks compared with that before continuous use, but no significance was observed in the Q group. The P group showed higher elasticity than the Q group, and was also significant.

Claims (1)

A hyaluronic acid production promoter to be blended in cosmetics containing N-acetylglucosamine, which contains chondroitin sulfate as an active ingredient, and a hyaluronic acid production promoter for human skin fibroblasts.