JP2005343882A - Hyaluronic acid production promoter, aquaporin synthesis promoter and composition of skin care preparation for external use - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a hyaluronic acid production promoter that is useful for preventing skin aging or treating diseases due to abnormal decomposition of hyaluronic acid by promoting hyaluronic acid production ability by cells and yet has slight influence on the human body and is safe, and a composition of a skin care preparation for external use, which fundamentally ameliorates skin function by increasing hyaluronic acid synthesis ability which cells themselves originally have and keeps skin firmness and moisture and prevents or ameliorates dry skin, tanned skin and aged skin by promoting aquaporin synthesis. <P>SOLUTION: The hyaluronic acid production promoter and the aquaporin synthesis promoter comprise an extract or essential oil obtained from Crocus sativus of the genus Crocus of the family Iridaceae. The composition of the skin care preparation for external use comprises the extract or essential oil obtained from the extract or essential oil obtained from Crocus sativus of the genus Crocus of the family Iridaceae, and N-acetylglucosamine. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a hyaluronic acid production promoter that promotes hyaluronic acid production in cultured cells or living bodies, and also relates to a hyaluronic acid production promoter that can be added to cosmetics, pharmaceuticals, etc. to enhance the hyaluronic acid production ability of the skin. The present invention also relates to an aquaporin synthesis promoter that is expected to have excellent moisture retention, rough skin improvement, aging prevention and the like by promoting aquaporin synthesis of skin cells, and the present invention further promotes the production of hyaluronic acid. Skin that can prevent or improve wrinkles, dry skin, tanned skin, and aging skin by containing N-acetylglucosamine, a substance having an action and aquaporin synthesis promoting action, and maintaining skin firmness and moisture The present invention relates to an external preparation composition.

  Hyaluronic acid retains moisture in the interstitial space, retains cells based on the formation of a jelly-like matrix in the tissue, maintains tissue lubricity and flexibility, resists external forces such as mechanical failure, and And has many functions such as prevention of bacterial infection (see Non-Patent Document 1).

  On the other hand, the hyaluronic acid staining intensity between epidermal cells decreases due to aging (see Non-Patent Document 2), and almost no hyaluronic acid is detected in the optical elasticity fibrosis due to ultraviolet irradiation (Non-Patent Documents 3 to 4). As a result, it is thought to cause dryness, firmness, loss of elasticity, and an increase in wrinkles. In order to improve such a state, a method of keeping the skin surface moisturized by applying a cosmetic containing hyaluronic acid has been taken, but since hyaluronic acid, which is a polymer, does not penetrate the skin, it is fundamentally Improvement cannot be expected. Therefore, it is expected to develop a substance that fundamentally improves the skin function by enhancing the hyaluronic acid synthesizing ability inherent in the cell itself.

  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 concentration of hyaluronic acid in normal human joint fluid is about 2.3 mg / mL, but in the case of rheumatoid arthritis, the concentration of hyaluronic acid in joint fluid decreases to about 1.2 mg / mL and at the same time the viscosity of joint fluid (See Non-Patent Document 5).

  In addition, it is known that hyaluronic acid content decreases in pyogenic arthritis and gouty arthritis as in the case of rheumatoid arthritis (see Non-Patent Document 6).

  In the above 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 rheumatoid arthritis patients are given joint injection therapy with sodium hyaluronate, the above improvement has been observed (see Non-Patent Document 7).

  Similarly, in the case of traumatic arthropathy, osteoarthritis and osteoarthritis, the above-described improvement effect has been reported by the joint injection therapy with hyaluronic acid (see Non-Patent Document 8).

  However, treatment of the above diseases is long-lasting and requires a doctor's prescription. Therefore, an ointment or gel 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. (See Non-Patent Document 9), hyaluronic acid production promoters are also expected as an early treatment for burns.

  Drugs that promote the production of hyaluronic acid in human cells include cytokines such as insulin-like growth factor-1 and epidermal growth factor (see Non-patent Document 10) and interleukin-1 (see Non-Patent Document 11), or phorbol esters ( Non-patent document 12) is known, but none of them can be used as a cosmetic, bathing agent or pharmaceutical product.

  In addition to recent studies showing that increasing hyaluronic acid has an excellent effect on the skin, aquaporin (hereinafter abbreviated as AQP) present in skin cells plays an important role in skin physiology. Became clear.

  AQP is also called a water channel, and is known as a membrane protein that selectively permeates water. So far, 11 types of AQP genes (AQP0 to AQP10) have been discovered in humans, and among them, AQP3, AQP7, and AQP9 permeate not only water but also nonionic small molecules such as glycerol and urea. In addition, when looking at the distribution of AQP expression in human tissues, it is clear that there are multiple AQPs in most organs.

Regarding the expression of AQP in the epidermis, recent expression studies by the present inventors have reported the expression of AQP3, AQP5, and AQP9 genes in cultured human keratinocytes and model skin epidermis. Among them, since expression of AQP3 in cultured human keratinocytes increases due to hyperosmotic stress, AQP3 is considered to be one important factor that controls the water environment of keratinocytes (see Non-Patent Document 13). In addition, in the skin of AQP3 gene-deficient mice, a decrease in stratum corneum water content, skin viscoelasticity, and barrier function recovery was observed, indicating that AQP3 is deeply involved in skin physical properties and skin physiological functions (Non-Patent Documents). 14). Therefore, the promotion of aquaporin production can be expected to suppress skin dryness, prepare a beam, and strengthen the barrier function. However, there is no known aquaporin synthesis promoter having a sufficient aquaporin synthesis promoting action, high safety, and good applicability to skin and the like.
“BIO INDUSTRY”, 1991, Vol. 8, p. 346 “J. Invest. Dermatol”, 1994, Vol. 102, p. 385 “Skin (increased by 5)”, 1997, Vol. 51, p. 53 “Nagoya Med. J.”, 1997, Vol. 41, p. 27 “Arthritis Rheumatism”, 1967, Vol. 10, p. 357 “Connective tissue”, 1984, Kanehara Publishing, p. 481 “Inflammation”, 1991, Vol. 11, p. 16 “Connective tissues and diseases”, 1980, Kodansha, p. 246 “Connective tissues and diseases”, 1980, Kodansha, p. 153 “Biochimica Biophysica Acta”, 1989, Vol. 1014, p. 305 “Journal of Japan Society of Obstetrics and Gynecology”, 1989, Volume 41, p. 1943 “Experimental Cell Research”, 1983, 148, p. 377 “Biochimica et Biophysica Acta”, 2001, vol. 1522, p. 82-88 “Journal of Dermatological Science”, 2002, Vol. 29, p. 143

  Therefore, the object of the present invention is to promote the hyaluronic acid production ability by cells, and thus can be used for the prevention of skin aging or the treatment of diseases accompanied by abnormal decomposition of hyaluronic acid, and it is safe and has little influence on the human body. It is in providing a hyaluronic acid production promoter.

  Another object of the present invention is to provide an aquaporin synthesis promoter that can be used for the purpose of improving moisture retention on skin, improving rough skin, and preventing aging by promoting aquaporin synthesis of skin cells. .

  In addition, the present invention can fundamentally improve the skin function by enhancing the hyaluronic acid synthesizing ability inherent in the cell itself, and further maintain the firmness and moisture of the skin by promoting aquaporin synthesis of the skin cell. Then, it aims at providing the skin external preparation composition which can prevent or improve a wrinkle, dry skin, sunburn skin, and aging skin.

  The above object is achieved by a hyaluronic acid production promoter and an aquaporin synthesis promoter characterized by containing an extract or an essential oil obtained from Crocus sativus, and further, Crocus sativus), an external preparation composition containing N-acetylglucosamine, an extract or essential oil obtained from skin, maintaining skin firmness and moisturizing, dry skin, tanned skin, aging It has been found that the effect of preventing or improving the skin is excellent.

  When the hyaluronic acid production promoter of the present invention [an extract or essential oil obtained from Crocus sativus] is added to a human skin fibroblast culture system, hyaluronic acid production is promoted in a concentration-dependent manner. (See Test Example-1 below). Therefore, the hyaluronic acid production promoter of the present invention acts on fibroblasts and can promote hyaluronic acid production in connective tissues such as skin, which are pathologically or physiologically reduced. In addition, an extract obtained from Crocus sativas exhibits an aquaporin synthesis promoting effect at a very low concentration (see Test Example 2 below). Also, by combining the hyaluronic acid production promoter and N-acetylglucosamine in combination, the amount of hyaluronic acid produced is increased, and aquaporin synthesis is further promoted to maintain excellent skin firmness and moisture. Thus, the effect of preventing or improving wrinkles, dry skin, tanned skin, and aging skin can be obtained.

  Hereinafter, the configuration of the present invention will be described in detail.

  Crocus sativus used in the present invention is a plant distributed in the northern hemisphere and gives purple flowers in autumn. The extract or essential oil obtained from this plant may be obtained from the whole flower or the female head. The extract can be obtained by a solvent extraction method, a liquefied gas extraction method such as liquefied carbon dioxide gas or liquefied butane gas, or a supercritical gas extraction method. Solvents used here include water or alcohols such as methanol, ethanol and isopropyl alcohol, polyhydric alcohols such as ethylene glycol, propylene glycol and 1,3-butylene glycol, ketones such as acetone, and esters such as ethyl acetate. , Ethers such as diethyl ether, aromatic compounds such as hexane, pentane and benzene can be selected from one kind alone or a mixture of two or more kinds. The essential oil can be obtained by the above method or steam distillation.

  The extract or essential oil obtained from Crocus sativus used in the present invention may be an extraction solvent solution or a steam distillate, or may be concentrated, dried and filtered by a conventional method. Re-extraction or the like may be performed.

  The hyaluronic acid production promoter of the present invention can be applied to cultured cells or living cells by itself to promote hyaluronic acid production. However, the hyaluronic acid production promoter of the present invention can be applied to skin external preparation compositions such as cosmetics and pharmaceutical compositions. You may mix | blend and use.

  The form of the hyaluronic acid production promoter, aquaporin synthesis promoter and the skin external preparation composition containing the hyaluronic acid production promoter of the present invention may be any of a liquid, a solid or a semisolid, and preferably an ointment, gel, cream, spray Agents, patches, lotions, packs, emulsions, powders, toiletries, sprouting agents, perfumes, bathing agents and the like can be mentioned, and the skin to be applied can be expected to be applied to all skin on the human body including the scalp.

  The hyaluronic acid production promoter and aquaporin synthesis promoter of the present invention can be combined with N-acetylglucosamine to increase hyaluronic acid production and promote aquaporin synthesis, resulting in excellent skin. A skin external preparation composition capable of preventing or improving wrinkles, dry skin, tanned skin, and aging skin while maintaining the firmness and moisture is obtained.

  N-acetylglucosamine used in the present invention is not limited to any product such as a synthetic product, a fermentation product, a degradation product obtained from chitin degradation such as crabs and shrimp.

  In addition to the above, the hyaluronic acid production promoter, aquaporin synthesis promoter and skin external preparation composition of the present invention include tar pigments, colored pigments such as iron oxide, preservatives such as paraben and phenoxyethanol, dimethylpolysiloxane. , Silicone oils such as methylphenylpolysiloxane, cyclic silicon, hydrocarbons such as paraffin and petrolatum, olive squalane, rice squalane, rice germ oil, jojoba oil, castor oil, safflower oil, olive oil, macadamia nut oil, sunflower oil, etc. Vegetable oils, waxes such as beeswax, owl, carnauba wax, octyldodecyl myristate, cetyl palmitate, isostearyl isostearate, ester oils such as isopropyl myristate, lower alcohols such as ethanol, cetanol, behenyl alcohol, stearyl alcohol, Higher alcohols such as chain-branched aliphatic alcohols, sterols and derivatives such as cholesterol, phytosterols, branched fatty acid cholesterol esters, macadamia nut fatty acid phytosteryl esters, processing oils such as hardened oils, stearic acid, myristic acid, isostearic acid, olein Higher fatty acids such as acids, iso-type long-chain fatty acids and anteiso-type long-chain fatty acids, terpenes such as limonene and hydrogenated bisabolol, glyceryl tricapryl / caprate, glyceryl 2-ethylhexanoate, glyceryl triiso-type long-chain fatty acid, tripalmitin Triglycerides such as glyceryl acid, sodium cetyl sulfate, anionic surfactants such as N-stearoyl-L-glutamate, polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, Nonionic surfactants such as reoxyethylene polyhydric alcohol fatty acid ester, polyoxyethylene hydrogenated castor oil, polyhydric alcohol fatty acid ester, polyglycerin fatty acid ester, modified silicon and sucrose ester, and cationic properties such as tetraalkylammonium salt Surfactant, amphoteric surfactant such as betaine type, sulfobetaine type, sulfoamino acid type, natural surfactant such as lecithin, lysophosphatidylcholine, ceramide, cerebroside, pigments such as titanium oxide, zinc oxide, dibutyl Antioxidants such as hydroxytoluene, sodium chloride, magnesium chloride, sodium sulfate, potassium nitrate, sodium sulfate, sodium metasilicate, calcium chloride and other inorganic salts, sodium citrate, potassium acetate, sodium oxalate, sodium aspartate Organic acids such as sodium lactate, dichloroacetic acid, mevalonic acid, glycyrrhizic acid and their salts, ethanolamine hydrochloride, ammonium nitrate, arginine hydrochloride, diisopropylamine salts, organic amines such as urea, decarboxycarnosine and their salts, edetic acid, etc. Chelating agent, xanthan gum, carboxyvinyl polymer, carrageenan, pectin, alkyl-modified carboxyvinyl polymer, thickener such as agar, neutralizing agent such as potassium hydroxide, diisopropanolamine, triethanolamine, hydroxymethoxybenzophenone sulfonate UV absorbers such as dipropylene glycol, 1,3-butylene glycol, glycerin, propylene glycol, sorbitol, malbitol, diglycerin, raffinose and other polyhydric alcohols, various alcohols Vitamins such as anoic acid, ascorbic acid, biotin, and tocopherol, and vitamin derivatives such as ascorbic acid sulfate ester salt, ascorbic acid phosphoric acid ester salt, and nicotinic acid tocopherol may be appropriately blended within the scope of achieving the object of the present invention it can.

  Furthermore, epidermal hyaluronic acid production promoters such as retinoids, keratinization promoters such as diisopropylamine dichloroacetic acid, niacin, mevalonic acid, hot spring water, sodium metasilicate, β-hydroxy-γ-aminobutyric acid, mevalonic acid, etc. The effect of preventing rough skin and wrinkles can be further enhanced by appropriately adding a moisturizer such as a barrier enhancer, hyaluronic acid, glucosamine, glucuronic acid, glycerol, urea, polyhydric alcohol, and the like.

  The content of extract or essential oil obtained from Crocus sativus used in the present invention in hyaluronic acid production promoter, aquaporin synthesis promoter and skin external preparation composition varies depending on its form. However, it is preferably 0.000001 to 0.01% (W / W) in terms of dry solid content, more preferably 0.00001 to 0.00. 001% (W / W). When the content is in the range of 0.000001 to 0.01% (W / W), the hyaluronic acid production effect and the aquaporin synthesis promoting effect are sufficiently obtained, and the touch at the time of use is good. The dosage form can be kept stable. That is, if the content is less than 0.000001%, the effect may not be sufficiently exhibited, and even if the content exceeds 0.01%, the effect commensurate with the content may not be obtained. However, in the case of a thing diluted at the time of use like a bath agent, the content can be further increased.

  When hyaluronic acid or aquaporin is produced in cultured cells, in the case of an extract or essential oil obtained from Crocus sativus, 0.0001% in terms of dry solids in the cell culture solution (W / W) or more is preferably contained, and more preferably 0.001% to 0.1%.

  The content of N-acetylglucosamine used in the present invention in the external preparation for skin is preferably 0.001 to 10% (W / W), particularly preferably based on the total amount of the external preparation for skin. Is 0.01 to 5% (W / W).

  Prior to the examples, test examples showing the effects of the present invention will be described. In addition, the preparation method and measurement method of the reagent used for each test are as follows.

-Cultured epidermal cells As human normal epidermal keratinocytes, commercially available products (Epidercell / newborn foreskin: manufactured by Kurabo Industries) were used.

-Epidermal cell culture medium Based on MCDB153HAA (purchased from Wako Pure Chemical Industries), hydrocortisone (0.5 μmol / L), ethanolamine (0.1 mmol / L), phosphoethanolamine (0.1 mmol / L), K-GM medium supplemented with insulin (5 μg / mL) and EGF (epidermal growth factor: 10 ng / mL) was prepared. For culture, 4 μL of BPE (bovine pituitary extract, purchased from Kurabo Corp.) was added to 1 mL of the medium to prepare a culture medium for epidermal cell culture.

Pronase solution 200 μg / mL Pronase (manufactured by Calbiochem-Bering Corporation, derived from Streptomyces griseus), 0.1 mol / L sodium chloride and 0.02% sodium azide-containing 0.5 mol / L Tris-HCl buffer (pH 8.0) ).

(Test Example 1) Hyaluronic acid production promoting action Test compound: Saffron extract [Saffron powder obtained from Crocus sativus was immersed in 99% ethanol, insoluble matter was filtered, and ethanol was removed (yield 18.9%)], and N-acetylglucosamine

2. Test method:
Cell culture The number of normal human epidermal cells was adjusted to 2.0 × 10 ⁴ cells / mL with a culture medium for epidermal cell culture, and 1.0 mL was seeded on a 24-well plate (Falcon). The culture was performed at 37 ° C. in an atmosphere of 95% (V / V) air-5% (V / V) carbon dioxide gas, and the sample was added on the fourth day of the culture. After further culturing for 3 days, the culture supernatant was recovered and the amount of hyaluronic acid in the supernatant was determined.

-Measurement of hyaluronic acid production amount Pronase solution 0.07mL was added to culture supernatant 0.63mL, and after leaving still at 37 degreeC for 18 hours, it heat-processed at 100 degreeC for 10 minutes, and inactivated pronase. Next, the amount of hyaluronic acid was measured from 0.01 mL of the reaction solution using a hyaluronic acid plate “Chugai” (manufactured by Chugai Diagnostics Science Co., Ltd.) according to the attached instructions. In addition, the sample addition tested in each group n = 3, and the result used the average value of each.

3. Test results: The amount of hyaluronic acid produced in the culture supernatant when various concentrations of saffron extract were added was measured (Table 1).
As a result, it was found that the amount of hyaluronic acid produced by epidermal cells increased in a dose-dependent manner by adding saffron extract.
Moreover, it turned out that hyaluronic acid production amount rises further by adding saffron extract and N-acetylglucosamine simultaneously. (Table 2)

Test Example 2 Aquaporin Synthesis Promotion Action Test compound: Saffron extract [Saffron powder obtained from Crocus sativus was immersed in 99% ethanol, insoluble matter was filtered, and then ethanol was removed (yield 18.9%)]

2. Test method:
-Sample addition and preparation of epidermal cell extract The number of human normal epidermal keratinocytes was adjusted to 1.0 × 10 ⁵ cells / mL with a culture medium for epidermal cell culture, and 3. 60 mm Petri dish (manufactured by Falcon) was used. 0 mL each was seeded. The culture was performed at 37 ° C. in an atmosphere of 95% (V / V) air-5% (V / V) carbon dioxide gas, and the sample was added on the fifth day of culture and further cultured for one day. After culturing, the medium is removed, washed with PBS, PBS is added again, and the cells are collected in a 1.5 mL tube with a cell scraper (manufactured by Sumitomo Bakelite), and the PBS is removed by centrifugation. Next, 500 μL of an extraction buffer [prepared by adding a complete miniprotease inhibitor cocktail tablet (Roche Diagnostics) to PBS] was added to the precipitate, followed by sonication [Handy Sonic UR-20P (TOMY SEIKO) )], And after centrifugation, the crude membrane fraction is further resuspended in elution buffer [SDS (sodium dodecyl sulfate) is prepared to a final concentration of 0.1% in the extraction buffer], and this is subjected to cell extraction. Liquid.

Western blotting (1) SDS polyacrylamide gel electrophoresis Cell extract 8 μL, 4 × concentrated SDS Sample Buffer (manufactured by Invitrogen) 4 μL, 10 × concentrated Sample Reding Agent (manufactured by Invitrogen) 1.6 μL, water 2.4 μL After mixing and heating at 70 ° C. for 10 minutes, it was subjected to electrophoresis. Electrophoresis was performed using 4-12% Bis-Tris Gel (manufactured by Invitrogen) according to the attached instructions of NuPAGE Gel system (manufactured by Invitrogen).

(2) Transfer of protein to PVDF membrane Transfer was performed using Xcell II Blot Module (manufactured by Invitrogen) according to the attached instructions. Immobilon (Millipore) was used as the PVDF membrane.

(3) Immunological detection The PVDF membrane after transfer was soaked in blocking reagent (reagent in which skim milk was dissolved in 5% concentration in PBS containing 0.05% Tween-20) for 30 minutes at room temperature, and then 3000 with blocking reagent. The mixture was reacted with human aquaporin-3 antibody diluted 2-fold at 4 ° C. for 24 hours. The human aquaporin-3 antibody was synthesized from the serum of a rabbit immunized with a C-terminal peptide of human aquaporin-3 amino acid sequence, and the dilution factor was set in advance to obtain an appropriate detection signal. . Next, the PVDF membrane was washed by shaking in PBS containing 0.05% Tween-20, and diluted with 2000% PBS containing 0.05% Tween-20 and anti-rabbit IgG antibody (manufactured by DAKO) at room temperature for 45 minutes. Reacted. Finally, the PVDF membrane was washed with shaking in PBS containing 0.05% Tween-20, and a signal was detected using SuperSignal (manufactured by PIERCE) according to the attached instructions. The results are shown in FIG. Tween-20 is a trade name of Uniqema Americas.

3. Test results Compared to the sample-free group (Comparative Experimental Example 1), a saffron extract 0.001% added group (Experimental Example 1), a 0.003% added group (Experimental Example 2), and a 0.01% added group (Experimental) In Example 3), a significant increase in detected signal intensity was observed. As is clear from this, it was confirmed that the saffron extract according to the present invention has an aquaporin synthesis promoting effect at an extremely low concentration.

  Examples of the present invention will be given below. In addition, the value in a table | surface shows the mass%. The saffron extract used in the examples is the same as that used in the above test examples.

Examples 1-4 (cream)
A cream was prepared with the composition shown below.

Preparation method: Component (A) was uniformly mixed and dissolved at 80 ° C., and then component (B) was mixed and dissolved therein (mixed solution I).
Separately, component (D) was uniformly mixed and dissolved at 80 ° C., and then component (C) was mixed and dissolved therein (mixed solution II).
Next, the liquid mixture II was gradually added to the liquid mixture I, and it cooled to 30 degreeC, fully stirring, and obtained the cream.

Examples 5-8 (Lotion)
A lotion was prepared with the composition shown below.

Preparation method: Each component was mixed and dissolved to prepare a lotion.

Examples 9 to 10 (baths)

Preparation method: Each component was mixed to prepare a bath agent. In addition, this bath agent is diluted about 3000 times at the time of use.

Examples 11-14 (ointment)

Preparation method: Each component of the above (B) is mixed in a hot water bath while heating to 80 ° C., and this is gradually added to the mixture of each component of (A) heated to 80 ° C. with stirring. It was.
Next, after vigorously stirring (2500 rpm) for 2.5 minutes with a homogenizer (manufactured by Tokuyukika Kogyou), each component was sufficiently emulsified and dispersed, and then gradually cooled with stirring to obtain an ointment.

Examples 18-21 (gel)

Preparation method: (A) was swollen with a part of water (D), component (C) was dissolved with the remaining water (D), and then both were uniformly mixed (mixed solution I).
Component (B) was mixed and dissolved uniformly (mixture II).
The mixture I was added to the mixture I and dispersed to obtain a gel.

Examples 19-22 (hair tonic)

Preparation method: After dissolving the fragrance with the fragrance solubilizer, the mixture was dissolved in ethanol while stirring at room temperature, and the component (B) was sequentially added and dissolved (mixed solution I).
The component (C) was dissolved, added to the mixture I with stirring, and leveled, and then filtered to obtain a hair tonic.

  It is a figure which shows the aquaporin synthesis promotion effect of the saffron extract in a human normal epidermal keratinocyte.

Claims (3)

  The hyaluronic acid production promoter characterized by containing the extract or essential oil obtained from Crocus sativas.   An aquaporin synthesis promoter characterized by containing an extract or essential oil obtained from Crocus sativus.   A skin external preparation composition comprising an extract or essential oil obtained from Crocus sativas and N-acetylglucosamine.

Images