WO2007075016A1 - Inhibiteur de collagenase contenant un complexe de vitamine c et d'acide poly-gamma-glutamique et utilisation correspondante - Google Patents

Inhibiteur de collagenase contenant un complexe de vitamine c et d'acide poly-gamma-glutamique et utilisation correspondante Download PDF

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WO2007075016A1
WO2007075016A1 PCT/KR2006/005684 KR2006005684W WO2007075016A1 WO 2007075016 A1 WO2007075016 A1 WO 2007075016A1 KR 2006005684 W KR2006005684 W KR 2006005684W WO 2007075016 A1 WO2007075016 A1 WO 2007075016A1
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pga
vitc
skin
conjugate
collagenase
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PCT/KR2006/005684
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English (en)
Inventor
Moon-Hee Sung
Chung Park
Jae-Chul Choi
Hiroshi Uyama
Haryoung Poo
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Bioleaders Corporation
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A61K8/676—Ascorbic acid, i.e. vitamin C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
    • A61K8/88—Polyamides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/52—Stabilizers
    • A61K2800/522—Antioxidants; Radical scavengers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74—Biological properties of particular ingredients
    • A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
    • A61K2800/782—Enzyme inhibitors; Enzyme antagonists

Definitions

  • the present invention relates to an inhibitor of collagenase, an enzyme that degrades collagen present in the dermis, the collagenase inhibitor containing a poly- gamma-glutamic acid- vitamin C complex (PGA- VitC complex or PGA- VitC conjugate), and to a composition for preventing skin wrinkles, which contains the PGA- VitC complex and can be used in drugs, cosmetics, foods and the like.
  • PGA- VitC complex poly- gamma-glutamic acid- vitamin C complex
  • PGA- VitC conjugate poly- gamma-glutamic acid- vitamin C complex
  • a composition for preventing skin wrinkles which contains the PGA- VitC complex and can be used in drugs, cosmetics, foods and the like.
  • UVA ultraviolet A
  • UVB ultraviolet B
  • UVC ultraviolet C
  • the dermal layer consists of a majority of type I collagen and a minor part of type III collagen, elastin, proteoglycan, fibronectin and the like (Bailly, C, J. Invest. Dermatol, 94:47, 1990).
  • elastin i.e., solar elastosis
  • proteoglycan increases, leading to a marked reduction in collagen, a major protein in dermis (Li, JJ. et al, Cancer Research, 56:483, 1996).
  • Collagen is a major structural protein produced in skin fibroblasts. It is an important protein present in the extracellular matrix and accounting for 30% of the total weight of body proteins and has a rigid triple-helical structure.
  • the main functions of collagen include providing mechanical firmness of the skin, cushioning connective tissue, the binding of tissue, maintenance of cell adhesion, and inducing cell division and differentiation. Particularly, it functions to impart strength and tension to the skin to protect the skin from external stimuli or forces.
  • the reduction of collagen that accounts for 90% of dermal volume was reported to have a very close connection with skin aging (Huang, C. et al, Proc. Natl. Acad. Sci. USA, 94:5826, 1997).
  • MMP matrix metalloproteinase
  • collagen has been used in cosmetics, foods, drugs and the like.
  • oligopeptides which is minimum active units of forming such collagen
  • peptides found to have a skin regeneration effect are known (US 4,665,054; WO 91/3488; WO 91/7431, etc.), but these peptides are known to have a problem in that a precipitate is formed, thus resulting in a great reduction in the stability of products.
  • Myofibroblasts are also known as ⁇ - smooth muscle actin-positive fibroblasts. When fibroblasts are irradiated with UVA, single oxygen is generated.
  • Reactive oxygen species derived from a singlet oxygen directly degrade or polymerize matrix components, which leads to the denaturation or fragmentation of the components, or cause a Maillard reaction, thus leading to the crosslinking of collagen fibers.
  • reactive oxygen species also function to activate collagenase, a collagen-degrading enzyme which is generally present in an inactive form.
  • the elimination of reactive oxygen species is effective for the prevention of skin aging through the inhibition of degradation or denaturation of matrix components or the inhibition of collagenase activity.
  • MMP matrix metalloproteinase
  • ECM extracellular matrix
  • BM base membrane
  • MT- MMP membrane-type MMP
  • MMP is a metalloproteinase having zinc in the active center thereof and is secreted in the form of zymogen from most cells, including skin keratinocyte and fibroblasts.
  • MMP is structurally modified, so that the amino-terminal end thereof is truncated and activated.
  • the activity of the activated MMP is regulated by inhibitors, such as ⁇ 2-macroglobulin or TIMPs (tissue inhibitors of metalloproteinase). Fisher et al.
  • collagenase a collagen- degrading enzyme
  • epidermal cells cells of tthe upper skin layer
  • reactive oxygen species reactive oxygen species
  • elastin an elastic fiber present between the collagen fibers
  • acts like a spring to maintain the tightness and firmness of the skin.
  • collagen is degraded by collagenase, and elastin is also crosslinked between chains to reduce skin elasticity, and thus the skin loses tightness and firmness and becomes wrinkled (FIG. 1).
  • retinoic acid Fisher, GJ. et al., Photochem. Photobiol, 69:154
  • TGF trans-forming growth factor
  • JP 8-231370 a protein derived from animal placenta
  • betulinic acid JP 8-208424
  • chlorella extracts JP 9-40523; JP 10-36283
  • the retinoic acid has limitations on its use, because it is unstable, causes severe irritation upon application to the skin and shows side effects, such as erythema, and has a stability problem.
  • the chlorella extracts and the like have an insignificant effect, and thus it is considered that these substances cannot provide the effect of improving skin functions by substantially promoting collagen synthesis of skin.
  • indole-3 -acetic acid is a plant growth hormone causing a tree to bear fruit or to bloom and is a substance, which regenerates the skin and inhibits collagenase that reduces the elasticity of the skin. It has received much attention as a raw material for functional cosmetics, but is known as a substance which is weak against light and heat and thus difficult to stabilize.
  • collagenase activity inhibitors including doxycycline, cartilage-derived molecules, Rb (retinoblastoma) (Chun, SJ. et al, Arthritis & Rheumatism, 50:78, 2004), triterpenoid-based substances, oleanolic acid, corsepin and its derivatives (Korean Patent Registration No. 10-0514315), and collagenase inhibitor containing ursolinic acid (Korean Patent Registration No. 10-0155613), are known, but mostly have problems in that they show side effects or are unstable in vivo.
  • composition useful for skin elasticity improvement and wrinkle prevention which has an inhibitory effect against collagenase, a collagen-degrading enzyme which is one major cause of skin aging, and, at the same time, can maintain stable activity in vivo and has no side effects.
  • PGA poly-gamma-glutamic acid
  • the present inventors have made extensive efforts to solve the above- described problems occurring in the prior art and, as a result, found that the poly- gamma-glutamic acid- VitC conjugate (complex) can maintain the elasticity of the skin by inhibiting collagenase degrading collagen, the component of skin connective tissue, and has high skin compatibility and an excellent moisture- absorbing effect, a moisturizing effect and a sustained-release effect, thereby completing the present invention.
  • Another object of the present invention is to provide a composition for preventing skin wrinkles, which contains the PGA- VitC conjugate as an active ingredient.
  • the present invention provides a collagenase inhibitor containing the PGA- VitC conjugate. Also, the present invention provides a composition for preventing skin wrinkles, which contains the PGA- VitC conjugate as an active ingredient.
  • FIG. 1 is a schematic diagram showing that collagen in the dermis of young skin is degraded by collagenase to become the dermis of old skin.
  • FIG. 2 is a graphic diagram showing the collagenase inhibitory effect of the PGA- VitC conjugate according to the present invention.
  • PGA-VC conjugate
  • PGA- VitC conjugate VC: VitC
  • PGA PGA- VitC conjugate
  • VC mixture a mixture of PGA and VitC.
  • FIG. 3 is a photograph showing the results of agarose gel analysis conducted to examine inhibition of MMP-I by PGA- VitC conjugate according to the present invention using cDNA prepared through reverse transcription-PCR (RT-PCR).
  • FIG. 4 is a bar graph showing the results of active MMP-I ELISA analysis conducted to examine inhibition of MMP-I by PGA- VitC conjugate according to the present invention.
  • the present invention relates to a collagenase inhibitor containing a PGA- VitC conjugate, and a composition for preventing skin wrinkles, containing the collagenase inhibitor.
  • the PGA preferably has a molecular weight of 1-15,000 kDa.
  • collagenase is preferably MMP-I, but the scope of the present invention is not limited thereto.
  • the composition for preventing skin wrinkles is preferably a cosmetic, pharmaceutical or food composition
  • the PGA- VitC conjugate is preferably contained in an amount of 0.01-60 wt%, and more preferably 0.1-50 wt%, based on the total weight of the composition, but the scope of the present invention is not limited thereto.
  • the inventive composition for preventing skin wrinkles, which inhibits collagenase can be used in the form of general external drug or functional cosmetic formulations.
  • the inventive composition can be formulated using general diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants and surfactants, or formulated by adding it to various cosmetic bases.
  • Liquid formulation for skin application include suspensions, internal solutions, and emulsions, and contain general simple diluents, including water and liquid paraffin, and various excipients, for example, wetting agents, flavoring agents, aromatics and preservatives, as well as various cosmetic bases.
  • the inventive composition for preventing skin wrinkles which contains the PGA-
  • VitC conjugate as an active ingredient can be used to prepare drugs, such as ointments for external skin application, lotions, such as astringent lotion, milk lotion and nourishing milk lotion, cosmetics, such as moisturizing cosmetics, cleansing foam, body cosmetics, lotions, creams, essence, and packs, and foods, including functional beverages, such as drinks, capsules or powder.
  • drugs such as ointments for external skin application
  • lotions such as astringent lotion, milk lotion and nourishing milk lotion
  • cosmetics such as moisturizing cosmetics, cleansing foam, body cosmetics, lotions, creams, essence, and packs
  • foods including functional beverages, such as drinks, capsules or powder.
  • functional beverages such as drinks, capsules or powder.
  • inventive composition can be prepared in any formulation, as long as it inhibits collagenase to provide an advantageous effect on skin elasticity improvement and wrinkle prevention.
  • the PGA used in the present invention is a viscous polymer consisting of D,L- glutamic acid bound to ⁇ -glutamyl, and is produced from Bacillus sp. strains isolated from, for example, Chungkookjang (Korean traditional fermented soybean food prepared using ricestraw), Natto (Japanese traditional fermented soybean food) and Kinema (Nepalese traditional fermented soybean food).
  • the PGA-VitC conjugate is an eatable, aqueous, anionic, biodegradable polymer material, which can be used as a moisture absorber, a moisturizer and a raw material for cosmetics.
  • the inventive PGA- VitC conjugate is a highly useful natural polymer, which can be used as low-water- soluble plastics, healthy foods having osteoporosis therapeutic effects, agents for reducing water contamination, biodegradable fibers or films and molded films, polymers for drug carriers, and the like.
  • a water-soluble powdery PGA- VitC conjugate was prepared by isolating PGA from Bacillus subtilis var chungkookjang (KCTC 0697BP) and treating the PGA with a gel containing VitC dissolved therein.
  • the effect of the PGA- VitC conjugate on collagenase inhibition was analyzed and, as a result, it could be seen that the effect of the PGA- VitC conjugate on collagenase inhibition was excellent compared to the cases where each of PGA, VitC and a mixture of PGA and VitC was added. Furthermore, the PGA- VitC conjugate had an excellent antioxidant effect, a sustained-release effect and a skin compatibility effect.
  • PGA was prepared according to the method described in the previous patent (WO 04/007593) filed in the name of the present inventors. That is, a culture broth of Bacillus subtilis var chungkookjang (KCTC 0697BP) was inoculated into a fermentor containing a basal medium to provide a PGA-containing sample solution. Then, the sample solution was left to stand to remove polysaccharides from the fermented solution, and the PGA precipitate was added with distilled water to dissolve. Next, protease was added to the solution and left to stand in incubator to degrade extracellular proteins present in the PGA sample, followed by dialyzing in a sufficient amount of distilled water to remove free glutamic acid, and concentrating to provide pure PGA.
  • KCTC 0697BP Bacillus subtilis var chungkookjang
  • the PGA thus obtained had an average molecular weight of 13,000 kDa, more than 95% of the PGA molecules had a molecular weight ranging from 3,000 to 15,000 kDa, and the PGA had a molecular weight distribution ranging from 1 kDa to 15,000 kDa.
  • a PGA- VitC conjugate (complex) was prepared according to the method descried in the previous patent (Korean Patent Registration No. 10-0498812) owned by the present inventors. That is, the PGA was placed in a dry test tube and dissolved in DMSO (dimethyl sufoxide), followed by stirring, thus obtaining transparent liquid. To the obtained transparent liquid, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) and N-hydroxysuccinimide were added, and stirred at room temperature, and then a DMSO solution containing VitC dissolved therein was added thereto, thus forming a gel.
  • DMSO dimethyl sufoxide
  • the PGA- VitC conjugate synthesized in Example 1 was dissolved at a concentration of 1%, and mixed with 400 unit/mL collagenase at collagenase concentrations of 0.002%, 0.003%, 0.004%, 0.006% and 0.008%, followed by reaction at 37 "C for 20 minutes. Meanwhile, 25 mg of bovine collagen was added to 0.5 mL of 0.05M TES (N-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid) buffer (pH 7.0) and then allowed to react at 37 ° C for 15 minutes. The collagen reaction solution was mixed with the collagenase reaction solution, and then allowed to react at 37 °C for 5 hours.
  • TES N-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid
  • Collagenase activity was calculated as the concentration of L-leucine liberated from collagen, and collagenase inhibition caused by PGA was calculated as the relative ratio of the enzymatic activity of collagenase, which reacted with PGA, to the enzymatic activity of collagenase, which did not react with PGA (Mandl, I. et ah, J. Clin. Invest, 32:1323, 1953).
  • FIG. 2 The results of measurement of collagenase inhibition caused by the PGA- VitC conjugate are shown in FIG. 2. As shown in FIG. 2, collagenase was strongly inhibited when the PGA- VitC conjugate was used, compared to when each of PGA, VitC and a mixture of PGA and VitC was used. Particularly, when the PGA-VitC conjugate was used at a concentration of 0.5 mM, collagenase inhibition reached about 90%.
  • Example 3 Cell assay for collagenase inhibition by PGA- VitC conjugate
  • a test on the degradation of collagen by each sample of PGA sample, VitC, a mixture of PGA and VitC and PGA- VitC conjugate was conducted in the following manner using human dermal fibroblasts by reverse transcription-PCR (RT-PCR), western blot analysis, and active MMP-I ELISA analysis.
  • Human dermal fibroblasts were cultured in a DMEM (dulbecco's modified eagle's medium) containing 10% FBS (fetal bovine serum), in a 5% CO 2 incubator at 37 ° C .
  • the cells used in the test were for 2-8 passages (1 passage is a period during which lxl ⁇ 5 cells grow into 1x10 6 cells).
  • 1 x 10 6 cells were inoculated into a 60 mm culture dish in the respective conditions and cultured for 18 hours before use in the test.
  • PGA poly-gamma- glutamic acid
  • VitC 0.05% PGA + 0.05% VitC
  • PGA- VitC conjugate each contained in 2 niL of PBS (phosphate buffered saline) buffer
  • PBS phosphate buffered saline
  • the media were collected and subjected to active MMP-I ELISA, and the cells were collected, from which mRNA was isolated and subjected to RT-PCR. Also, protein lysates were collected and subjected to western blot.
  • RT-PCR analysis of MMP-I inhibition For RT-PCR, the culture broth was removed from the cells inoculated into the 60 mm culture dish, and the cells were repeatedly pipetted with 1 mL of TRIZOL (Invitrogen Corp.) solution. Then, the solution in the culture dish was collected into a 1.5 mL tube and allowed to react at room temperature for 5 minutes. 0.2 mL of chloroform was added to the reaction sample, and the mixture was strongly vortexed and then allowed to react at room temperature for 5 minutes. After the reaction, the reaction solution was centrifuged at 4 ° C and 13,000 rpm for 15 minutes, and the supernatant was isolated.
  • TRIZOL Invitrogen Corp.
  • the isolated supernatant was collected into a 1.5 mL fresh tube, 0.5 mL of isopropyl alcohol was added thereto to mix, and the mixture was allowed to react at room temperature for 10 minutes. After the reaction, the reaction solution was centrifuged again at 4°C and 13,000 rpm for 10 minutes, and the supernatant was removed. 1 mL of 75% ethanol (EtOH) was added thereto and vortexed, and the solution was centrifuged at 4 ° C and 13,000 rpm for 20 minutes. The supernatant was removed, and the precipitate was dried at room temperature for 10 minutes. Then, DEPC (diethyl pyrocarbonate) solution water was dropped onto the RNA precipitate, and the RNA was quantified using spectrophotometry.
  • EtOH 75% ethanol
  • RNA 2 ⁇ g of the quantified RNA was placed in a PCR tube, 1 ⁇ L of an oligo-dT random primer was added into the tube, and DEPC distilled water was added thereto to a final volume of 15 ⁇ L.
  • the above-prepared mixture was allowed to react at 70 °C for 5 minutes, and then 10 ⁇ L of a buffer mixture, 2 ⁇ L of M-MLV (moloney murine leukemia virus) 5 x reaction buffer, 1.25 ⁇ L of 10 mM dNTP, 10 units of RNaseOUT (Invitrogen Corp.).
  • M-MLV moleukemia virus
  • MMP matrix metalloproteinase
  • PCR reaction was performed in the following conditions: pre-denaturation at 95 °C for 5 min, and then 30 cycles, each consisting of 40 sec at 95 ° C , 40 sec at 55 °C and 80 sec at 72 ° C , followed by extension at 72 ° C for 5 min.
  • the constructed cDNA was electrophoresed on 1.0% agarose gel containing ethidium bromide (EtBr), and the density of each band was measured using a densitometry (see FIG. 3).
  • EtBr ethidium bromide
  • Cl represents a control group non-irradiated with UVA
  • C2 represents a control group irradiated with UVA.
  • ⁇ — actin antibody As shown in FIG.
  • An active MMP-I ELISA test was carried out using the prepared culture broth and human active MMP-I fluorescent assay kit of R&D system (FlMOO).
  • a working standard solution and a sample solution were previously prepared and 100 ⁇ L of assay diluent RD1-64 was added into each well.
  • 150 ⁇ L of each of the working standard solution and the sample solution was added into each well, and the mixture was allowed to react at room temperature for 3 hours. Then, the reaction solution was removed, and each well was washed four times with 400 ⁇ L of wash buffer using a multi-channel pipette.
  • Free radical removing ability of the PGA- VitC conjugate prepared in Example 1 was examined.
  • the used DPPH l,l-diphenyl-2-picrylhydrazyl
  • the radical removing ability of the PGA- VitC conjugate was examined by measuring O.D. value.
  • the PGA-VitC conjugate was dissolved in distilled water or solvent, and 100 ⁇ L of the liquid sample was mixed with 100 ⁇ L of 200 ⁇ M DPPH. The mixture solution was left to stand in a dark chamber at room temperature for 10 minutes and measured for absorbance at A518 nm in comparison with PGA, VitC and the PG A/ VitC mixture as control groups. Also, EDA (electron donating ability) was calculated from an equation of (absorbance in control group - absorbance in test group)/absorbance in control group x 100 (see Table 1).
  • Example 5 Sustained-release effect of PGA- VitC conjugate
  • mice thirty 4-week-old Balb/c mice were bred in a mouse cage with a 12-hr light/ 12-hr dark cycle while the animals were given free access to feed and distilled water.
  • the mice were anesthetized with ether, the entire small intestine region from the duodenum to the ileum was removed from the abdomens of the mice, and was divided into the upper and lower parts, the contents of which were washed out with cold physiological saline. Then, the small intestine tissue was homogenized with a homogenizer while suitable cold physiological saline was added thereto.
  • the homogenized small intestine tissue was centrifuged at 4 ° C and 8,000 x g for 20 minutes, and the soluble part and non-soluble part of each fraction were separated from each other and stored at -20 ° C , while VitC contained therein was analyzed by HPLC.
  • the skin safety of a cosmetic composition containing the PGA- VitC conjugate was measured. Specifically, 30 volunteers (average age: 25 years old; age distribution: 19-40 years old) were divided into two groups A and B. The groups A and B were subjected to a skin patch test with the inventive cosmetic composition and a comparative cosmetic composition, respectively, using the Haye's Test Chamber. Herein, persons who had skin lesions, such as psoriasis or eczema, pregnant women and nursing mothers, or persons who were being administered with antihistamine agents, were excluded from the test. The test site was washed with 70% ethanol and dried. Next, for each person of the groups A and B, 15 ⁇ g of each of the samples was dropped into the chamber, and then fixed onto the upper arm site as the test site.
  • the patch was applied for 24 hours, and after the patch was removed, the test site was marked with a marking pen. At 24 hours, 48 hours and 72 hours, the test site was observed, and evaluated according to the criteria of International Contact Dermatitis Research Group (ICDRG) (see Table 2 and Table 3).
  • IDRG International Contact Dermatitis Research Group
  • formulation examples of the present invention skin lotion, milk lotion, essence, cleaning foam and shampoo will be illustrated as formulation examples of the present invention, but formulations containing the inventive cosmetic composition are not limited thereto.
  • the PGA- VitC conjugate prepared in Example 1 butylene glycol, glycerin, a carboxyvinyl polymer, arginine, a preservative and purified water were mixed with each other and heated at 70-75 ° C with stirring. Meanwhile, squalane, butylene glycol dicaprylate/dicaprate, sorbitan stearate, polysorbate 60, glyceryl stearate and stearyl glycyrrhetinate were mixed with each other and heated at 75-80 °C with stirring, and the mixture was added to the above mixture containing the PGA- VitC conjugate, to form an emulsion.
  • Sitosterol, polyglyceryl 2-oleate, ceramide, ceteares-4 and cholesterol were mixed with each other with stirring. Then, to the mixture, a mixture solution of the PGA- VitC conjugate, dicetyl phosphate, concentrated glycerin and purified water was added to form an emulsion. The emulsion was cooled to 45 ° C with stirring, and a fragrance was then added thereto. Then, the mixture was stirred, cooled to 30 ° C and aged. Then, the mixture was stabilized by adding a carboxyvinyl polymer, xantan gum and a preservative thereto, and then aged, thus preparing an essence containing the PGA- VitC conjugate. The content of each component in the essence is shown in Table 6 below. Table 6
  • N-acylglutamic acid sodium salt, glycerin, PEG-400 and propylene glycol were mixed with each other in purified water, and the PGA- VitC conjugate was added thereto in small portions. Then, EDTA-4Na was added thereto, and the mixture was heated at 80 ° C with stirring to dissolve. Meanwhile, POE(15) oleylalcohol ether, lauryl derivatives and methyl paraben were mixed and heated at 80 ° C , and the mixture was added to the mixture containing the PGA- VitC conjugate. The resulting mixture was stirred, and a fragrance was added thereto. Then, the mixture was slowly cooled, thus affording a cleansing foam containing the PGA- VitConjugate. The content of each component in the cleansing foam is shown in Table 7 below. Table 7
  • the present invention provides the collagenase activity inhibitor containing the poly-gamma-glutamic acid (PGA)-VitC conjugate, and the composition for preventing skin wrinkles, which contains the PGA- VitC conjugate.
  • the collagenase inhibitor according to the present invention has not only the effect of inhibiting collagenase activity, but also antiaging effects, such as the effect of inhibiting matrix metalloproteinase to provide an antioxidant effect and reduce skin wrinkles.
  • the collagenase inhibitor maintains the elasticity of the skin by keeping skin connective tissue taut and has high skin compatibility, moisturizing ability and moisture-absorbing ability. Thus, it is useful to provide cosmetic, pharmaceutical and food compositions for improving skin elasticity and reducing wrinkles.

Abstract

L'invention concerne un inhibiteur de collagénase contenant un complexe de vitamine C et d'acide poly-gamma-glutamique (conjugué de PGA-VitC), et une composition destinée à prévenir les rides, qui contient le conjugué de PGA-VitC et qui peut être utilisé dans des médicaments, des produits cosmétiques et des aliments. Le conjugué de PGA-VitC possède non seulement l'effet inhibiteur de l'activité de la collagénase, mais aussi les effets antivieillissements, notamment un effet antioxydant et un effet améliorant les rides cutanées. En outre, l'inhibiteur de collagénase entretient l'élasticité de la peau en maintenant tendus les tissus conjonctifs cutanés et présente une compatibilité dermique élevée, des effets hydratants, absorbant l'humidité et de libération soutenue excellents. Il est donc utile de produire des compositions cosmétiques, pharmaceutiques et alimentaires en vue d'améliorer l'élasticité de la peau et de réduire les rides cutanées.
PCT/KR2006/005684 2005-12-29 2006-12-22 Inhibiteur de collagenase contenant un complexe de vitamine c et d'acide poly-gamma-glutamique et utilisation correspondante WO2007075016A1 (fr)

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2153814A1 (fr) 2008-08-05 2010-02-17 Isdin S.A. Utilisation de composition comportant de l'urée
EP2153815A1 (fr) 2008-08-05 2010-02-17 Isdin S.A. Utilisation de compositions contenant de l'urée
EP2415468A1 (fr) * 2009-03-30 2012-02-08 Shiseido Company, Ltd. Composition atténuant les lésions causées par les rayons ultraviolets
EP2484356A1 (fr) * 2009-09-30 2012-08-08 Shiseido Company, Ltd. Composition orale pour réduire la formation de rides
EP2484357A1 (fr) * 2009-09-30 2012-08-08 Shiseido Company, Ltd. Composition orale permettant de réduire la rugosité cutanée
CN102652835A (zh) * 2010-12-10 2012-09-05 邹长坪 缓释高分子维甲酸
FR3077706A1 (fr) * 2018-02-15 2019-08-16 Mixscience Composition contre les stress
CN113046397A (zh) * 2021-03-30 2021-06-29 山东丰金生物工程有限公司 一种低分子量6-o-pga-l-抗坏血酸的非水相酶法合成方法
CN113087899A (zh) * 2021-03-30 2021-07-09 山东丰金生物工程有限公司 一种l-抗坏血酸衍生物及其制备方法、应用

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KR102017741B1 (ko) * 2017-12-01 2019-09-03 주식회사 리엔젠 비타민 c 유도체를 고정화한 폴리감마 글루탐산계 진피 충전제 조성물 및 그 제조방법

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KR100485727B1 (ko) * 2004-06-24 2005-04-27 주식회사 바이오리더스 폴리감마글루탐산-비타민 복합체 및 그 용도

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KR100485727B1 (ko) * 2004-06-24 2005-04-27 주식회사 바이오리더스 폴리감마글루탐산-비타민 복합체 및 그 용도

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2153814A1 (fr) 2008-08-05 2010-02-17 Isdin S.A. Utilisation de composition comportant de l'urée
EP2153815A1 (fr) 2008-08-05 2010-02-17 Isdin S.A. Utilisation de compositions contenant de l'urée
EP2415468A1 (fr) * 2009-03-30 2012-02-08 Shiseido Company, Ltd. Composition atténuant les lésions causées par les rayons ultraviolets
EP2415468A4 (fr) * 2009-03-30 2014-08-20 Shiseido Co Ltd Composition atténuant les lésions causées par les rayons ultraviolets
EP2484357A4 (fr) * 2009-09-30 2013-11-20 Shiseido Co Ltd Composition orale permettant de réduire la rugosité cutanée
EP2484357A1 (fr) * 2009-09-30 2012-08-08 Shiseido Company, Ltd. Composition orale permettant de réduire la rugosité cutanée
EP2484356A4 (fr) * 2009-09-30 2013-12-04 Shiseido Co Ltd Composition orale pour réduire la formation de rides
EP2484356A1 (fr) * 2009-09-30 2012-08-08 Shiseido Company, Ltd. Composition orale pour réduire la formation de rides
CN102652835A (zh) * 2010-12-10 2012-09-05 邹长坪 缓释高分子维甲酸
FR3077706A1 (fr) * 2018-02-15 2019-08-16 Mixscience Composition contre les stress
WO2019158736A1 (fr) * 2018-02-15 2019-08-22 Mixscience Composition contre les stress
CN113046397A (zh) * 2021-03-30 2021-06-29 山东丰金生物工程有限公司 一种低分子量6-o-pga-l-抗坏血酸的非水相酶法合成方法
CN113087899A (zh) * 2021-03-30 2021-07-09 山东丰金生物工程有限公司 一种l-抗坏血酸衍生物及其制备方法、应用
CN113087899B (zh) * 2021-03-30 2023-03-07 山东丰金生物工程有限公司 一种l-抗坏血酸衍生物及其制备方法、应用

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