WO2013137505A1 - 신규한 trpv1 억제 펩티드 및 이를 함유하는 피부노화 방지 또는 주름 개선용 조성물 - Google Patents
신규한 trpv1 억제 펩티드 및 이를 함유하는 피부노화 방지 또는 주름 개선용 조성물 Download PDFInfo
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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
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
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- 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
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/04—Antipruritics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- 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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
Definitions
- the present invention relates to an activity inhibitory peptide of TRPV1 affecting intracellular Ca 2+ influx, and a composition for preventing skin aging, improving skin wrinkles, skin whitening, inflammation, itching, or pain.
- Skin aging is often caused by external and internal (natural) processes associated with an increase in wrinkles, sagging and sagging, and is generally referred to as 'photoaging' because external aging is primarily caused by repeated ultraviolet rays (UV) exposure. It is called.
- Naturally aged skin is smooth, pale and fine wrinkles, but photo-aged skin develops thick wrinkles and causes pigmentation and capillary dilatation.
- MMP matrix metalloproteinases
- transient receptor potential vanilloid type 1 is one of a family of calcium-permeable non-selective cation channels, in which TRPV1 activity induces Ca 2+ uptake and is inhibited by certain antagonists such as capsazepine. .
- TRPV1 is directly activated by capsaicin, heat, low pH, bradykinin, PGE 2 , or ATP.
- TRPV1 is the primary biological sensor for thermo-chemical stimulation and tissue damage.
- TRPV1 is reported to be present in various tissues such as brain, kidney, bronchial epithelial cells and epidermal keratinocytes.
- capsaicin stimulation increases the concentration of Ca 2+ in the cytoplasm of keratinocytes, which are inhibited by capsazepine.
- TRPV1 The role of TRPV1 in the heat induced MMP-1 expression of epidermal cells has been reported, and UV light is a more decisive factor in inducing MMP expression and skin aging.
- Ca 2+ influx through TRPV1 is a critical factor in UV-induced MMP-1 expression, and Ca 2+ dependent protein kinase C (PKC) is involved in signal transduction.
- PKC Ca 2+ dependent protein kinase C
- epidermal TRPV1 will function as a sensor for harmful stimuli such as ultraviolet light. Therefore, since TRPV1 may be a target for preventing skin photoaging caused by ultraviolet light exposure, there is a need for discovering a novel TRPV1 activity inhibiting substance in order to prevent and prevent skin aging.
- An object of the present invention is to provide a TRPV1 inhibitory peptide and a composition for preventing skin aging, improving skin wrinkles, skin whitening, inflammation, itching, or pain.
- the present invention provides a peptide described in SEQ ID NO: 1 to SEQ ID NO: 8.
- the peptide is identified as a novel TRPV1 activity inhibitory peptide.
- the TRPV1 is a non-specific cation channel belonging to the TRP family and is a membrane protein that is activated by capsaicin, a hot ingredient of red pepper, induces cation and forms a membrane voltage, thereby transmitting stimuli to the nervous system.
- the activity of TRPV1 is regulated through several pathways, and it is known that activity is regulated by phosphorylation and dephosphorylation including PKA, PKC, Ca 2+ / calmodulin-dependent protein kinase (CaMK) and Src kinase. It may also be regulated by post-transcriptional regulatory mechanisms in addition to phosphorylation.
- PKA PKA
- PKC Ca 2+ / calmodulin-dependent protein kinase
- Src kinase Src kinase
- the novel peptides described in SEQ ID NOS: 1-8 have been identified.
- the peptide is a peptide capable of inhibiting the CaMK activity site that affects intracellular Ca 2+ influx by TRPV1 activity or a phosphorylated amino acid-based peptide sequence of TRPV1 protein, and thus it was confirmed that the peptide effectively inhibits TRPV1 activity.
- One peptide can be usefully used as an inhibitor of TRPV1 activity.
- the present invention also provides a cosmetic composition for preventing skin aging or for improving wrinkles, which contains any one or more selected from the group consisting of peptides represented by SEQ ID NOs: 1 to 8.
- the anti-aging of the skin refers to the action of inhibiting skin aging due to photoaging or natural aging, but the skin aging is preferably photoaging due to UV exposure.
- the skin wrinkles may be due to photoaging or natural aging, more preferably by photoaging, and more preferably, photoaging by UV exposure, but is not limited thereto.
- the skin aging and wrinkles are preferably based on TRPV1 activity, but is not limited thereto.
- the peptide is preferably contained at a concentration of 0.001 to 20 mM in the cosmetic composition, more preferably at a concentration of 0.01 to 10 mM, but is not limited thereto.
- concentration of the peptide is less than 0.001 mM, TRPV1 inhibition and anti-aging and skin wrinkle improvement effects cannot be obtained.
- concentration of the peptide exceeds 20 mM, there is a problem in that production economy is inferior due to no apparent increase in effect due to an increase in content.
- the peptide of the present invention can be prepared by general chemical synthesis, for example, solid-phase peptide synthesis, and by culturing a microorganism transformed with a recombinant vector containing a nucleic acid encoding the peptide.
- the peptide may be expressed and then purified by conventional methods, but is not limited thereto.
- the cosmetic composition according to the present invention may further contain, in addition to the above peptides, other components that can give a synergistic effect to the effects of the peptides within the range of not impairing the effects aimed at by the present invention.
- other components that can give a synergistic effect to the effects of the peptides within the range of not impairing the effects aimed at by the present invention.
- conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings, or carriers.
- the cosmetic composition of the present invention is not particularly limited in its formulation, and may be prepared in any formulation conventionally prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, And creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, sprays and the like. More specifically, it may be prepared in the form of a lotion, essence, lotion, cream, pack, gel, ointment, patch or spray.
- the formulation of the cosmetic composition is a paste, cream or gel
- animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components.
- Lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as the carrier component in the case of powders or sprays, in particular chlorofluorohydrocarbons, propane / Propellants, such as butane or dimethylether.
- a solvent, solubilizer or emulsion may be used as the carrier component, for example water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Fatty acid esters of propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.
- liquid carrier diluents such as water, ethanol or propylene glycol
- suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals
- ethoxylated isostearyl alcohol such as water, ethanol or propylene glycol
- suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters
- microcrystals Cellulose, aluminum meta hydroxide, bentonite, agar or tracant and the like
- surfactant-containing cleansing as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate , Imidazolinium derivatives, methyltaurate, sarcosinates, fatty acid
- TRPV1 is involved in the regulation of skin proliferation and differentiation, and TRPV1 mediated calcium influx in cultured human keratinocytes has been reported to inhibit cell proliferation and induce cell death. Moreover, activation of TRPV1 by capsaicin or heat has been known to alter the formation of the permeation barrier of the epithelium in human skin.
- the present inventors have studied to find a novel inhibitor that inhibits the activity of TRPV1 as described above, the novel TRPV1 inhibitory peptide (peptide 1: QRRPSLKSL, peptide 2: QRAITILDT, peptide 3: RRPSL, Peptide 4: RAITI, Peptide 5: MHRQETVDC, Peptide 6: LKKFNARRKL, Peptide 7: RQETV and Peptide 8: KFNAR) were established and the effect of inhibiting skin aging and wrinkle improvement by the peptide was established.
- the novel TRPV1 inhibitory peptide peptide 1: QRRPSLKSL, peptide 2: QRAITILDT, peptide 3: RRPSL, Peptide 4: RAITI, Peptide 5: MHRQETVDC, Peptide 6: LKKFNARRKL, Peptide 7: RQETV and Peptide 8: KFNAR
- peptide to HaCaT cells which are human keratinocytes
- expression of MMP-1 induced by UV exposure was reduced (see FIGS. 1A and 1B)
- IL The expression of pro-inflammatory cytokines such as -1 ⁇ , IL-6, IL-8 and TNF- ⁇ has been shown to be inhibited (see FIGS. 2A-2H), thereby allowing peptides of the invention to be exposed to UV radiation. It was found that it suppresses the increased intracellular phenomenon.
- capsaicin which is an activation inducer of TRPV1
- peptide treatment after capsaicin was shown to inhibit intracellular Ca 2+ influx by capsaicin (see FIG. 3).
- the skin of hairless mice was irradiated with UV and treated with peptides (see FIG. 4).
- the peptides concentration-dependently reduced the skin thickness increased by UV exposure see FIGS. 5 and 9
- MMP-13 and MMP-9 see FIGS. 6 and 7
- Procollagen, a precursor of collagen was shown to increase (see FIG. 8).
- the effect of inhibiting apoptosis induced by UV exposure was confirmed (see FIG. 10).
- the novel peptide of the present invention has the effect of inhibiting the skin aging phenomenon induced after UV exposure by inhibiting the activity of TRPV1 involved in skin aging, inhibiting the expression and activity of MMP and improving the expression of procollagen. Since it can contribute to skin wrinkle and elasticity improvement, the peptide can be usefully used as an active ingredient of the cosmetic composition for preventing skin aging and wrinkle improvement.
- the present invention also provides a pharmaceutical composition for preventing skin aging or improving wrinkles, which contains any one or more selected from the group consisting of peptides represented by SEQ ID NOs: 1 to 8.
- the skin aging may be photo aging or natural aging, more preferably photo aging, the photo aging is most preferably due to UV exposure, but is not limited thereto.
- the concentration of the peptide contained in the pharmaceutical composition is preferably 0.001 to 20 mM, more preferably 0.01 to 10 mM, but is not limited thereto. If the concentration of the peptide is less than 0.001 mM, the effect of preventing skin aging or wrinkle improvement may not be obtained. If the concentration of the peptide exceeds 20 mM, there is a problem of using more peptide than necessary.
- TRPV1 inhibitory peptide of the present invention inhibits the expression of MMPs and proinflammatory cytokines induced by UV exposure in human keratinocytes and in vivo, reduces skin thickness and apoptosis, and reduces procollagen with UV exposure Since it increases the expression of, it can be usefully used as an active ingredient of the pharmaceutical composition for preventing skin aging or improving wrinkles.
- the present invention also provides a pharmaceutical composition for skin whitening containing any one or more selected from the group consisting of peptides set forth in SEQ ID NO: 1 to SEQ ID NO: 8 as an active ingredient.
- the concentration of the peptide contained in the pharmaceutical composition is preferably 0.001 to 20 mM, more preferably 0.01 to 10 mM, but is not limited thereto. If the concentration of the peptide is less than 0.001 mM, the skin whitening effect may not be sufficiently obtained. If the concentration of the peptide exceeds 20 mM, the skin whitening effect does not increase relative to the concentration.
- the TRPV1 inhibitory peptides of the invention inhibit the expression of MMP induced by UV exposure in human keratinocytes and in vivo, reduce skin thickness and apoptosis, and inhibit the expression of procollagen. Increase. Since the TRPV1 inhibitory peptide shows an effect on skin blackening and skin whitening activity by UV exposure through the proliferation and regeneration of these keratinocytes, it can be usefully used as an active ingredient of a pharmaceutical composition for skin whitening.
- the present invention provides a pharmaceutical composition for improving inflammation, itching, or pain containing any one or more selected from the group consisting of peptides set forth in SEQ ID NO: 1 to SEQ ID NO: 8 as an active ingredient.
- the concentration of the peptide contained in the pharmaceutical composition is preferably 0.001 to 20 mM, more preferably 0.01 to 10 mM, but is not limited thereto.
- the inflammation, itching and pain may be caused by stimulation by UV exposure, but is not limited thereto.
- the inhibitory effect of the TRPV1 inhibitory peptide on pro-inflammatory cytokines induced by UV exposure in human keratinocytes and in vivo As a result, the expression levels of proinflammatory cytokines IL-1 ⁇ , IL-6, IL-8 and TNF- ⁇ were found to be significantly reduced (see FIGS. 2A to 2H).
- the TRPV1 has been reported to cause inflammation in various diseases and injuries.
- TRVP1 is known to promote the secretion of proinflammatory cytokines by inducing activity by capsaicin in keratinocytes derived from human epithelium and hair follicles, and has been known to be associated with TRPV1 and inflammation.
- TRPV1 knockout mice have increased heat thresholds and reduced tissue swelling in arthritis (see Pharmacology and Therapeutics 2010; 125: 181.95, Science 2000; 288: 306.13, Arthritis and Rheumatism 2005; 52: 3248.56).
- TRPV1 plays an important role in the pain delivery pathway. That is, when TRPV1 is activated by a pain medium, cations are introduced through the cation channel TRPV1, and pain is transmitted to the central nervous system by the action potential. Therefore, TRPV1 has already been studied as an important target molecule for analgesic and anti-inflammatory development (see Journal of the Korea Academia-Industrial cooperation Society, 2011, pp. 3096-3102).
- TRPV1 is associated with the development of skin itch, which has been shown to occur through itching-specific subpopulation of sensory afferent neurons expressing TRPV1 ( J. Clin. Invest. 116, 11741186, 2006, Biochim. Biophys.Acta 1772, 10041021, 2007). TRPV1 is expressed in human skin and not in neuronal cells, particularly in epidermal keratinocytes of patients suffering from nodular itch rash.
- the TRPV1 inhibitory peptide of the present invention can improve inflammation, itching or pain based on increasing TRPV1 activity by inhibiting TRPV1 activity, and thus can be usefully used as an active ingredient of an inflammation, itching or pain pharmaceutical composition.
- Routes of administration of the pharmaceutical compositions according to the invention include oral, intravenous, intramuscular, intraarterial, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal, with oral or parenteral administration being preferred.
- oral or parenteral administration is preferred.
- the parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intralesional injection or infusion techniques.
- composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the TRPV1 inhibitory peptide of the present invention.
- the composition may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.
- Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills and the like.
- Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.
- Formulations for parenteral administration include powders, granules, tablets, capsules, sterile aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc.
- an external skin pharmaceutical composition of cream, gel, patch, spray, ointment, warning agent, lotion agent, linen agent, pasta agent or cataplasma agent may be prepared and used. It is not limited to this.
- the non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
- the base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
- the composition may further contain preservatives, stabilizers, emulsifiers or emulsifiers, auxiliaries such as salts and / or buffers for the control of osmotic pressure, and other therapeutically valuable substances and may be mixed, granulated or It may be formulated according to the coating method.
- the pharmaceutical composition of the present invention may vary depending on several factors, including the age, weight, general health, sex, time of administration, route of administration, rate of release, drug combination and severity of the particular disease of the individual.
- the peptide contains the peptide of the present invention as an active ingredient in a unit dose of about 0.01 to 1,500 mg, Dosages required range from about 1 to 500 mg per day, depending on the frequency and intensity of administration, but are not limited thereto. For intramuscular or intravenous administration to adults, a full dose of about 5 to 300 mg per day will usually be sufficient, but for some patients a higher daily dose may be desirable.
- composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.
- the present invention provides a method for preventing skin aging or improving wrinkles, comprising the step of treating or administering to a subject any one or more selected from the group consisting of peptides described in SEQ ID NO: 1 to SEQ ID NO: 8 in a pharmaceutically effective amount. to provide.
- the present invention also provides a skin whitening method comprising the step of treating or administering to a subject any one or more selected from the group consisting of peptides described in SEQ ID NOs: 1 to 8.
- the present invention is a method for improving inflammation, itching, or pain, comprising the step of treating or administering to a subject any one or more selected from the group consisting of peptides set forth in SEQ ID NO: 1 to SEQ ID NO: 8 in a pharmaceutically effective amount To provide.
- the pharmaceutically effective amount is 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, but is not limited thereto.
- the treatment or dosage may be adjusted according to the weight, age, sex, health condition, diet, duration of administration, method of administration, clearance, severity of the disease, etc. of the particular patient. Treatment or administration may be administered once a day or may be divided several times.
- the subject is a vertebrate and preferably a mammal, more preferably an experimental animal such as a rat, rabbit, guinea peak, hamster, dog or cat, most preferably an ape-like animal such as a chimpanzee or gorilla.
- an experimental animal such as a rat, rabbit, guinea peak, hamster, dog or cat
- an ape-like animal such as a chimpanzee or gorilla.
- the treatment or method of administration may be applied, oral or parenteral, and may be intraperitoneal, intrarectal, subcutaneous, intravenous, intramuscular, intrauterine dural, cerebral vascular or thoracic It can be administered by internal injection.
- the inflammation is dermatitis, allergy, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia , Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It may be one, but is not limited thereto.
- the TRPV1 inhibitory peptide of the present invention effectively inhibits the activity of TRPV1, it can be usefully used for alleviating and improving skin aging, wrinkles, whitening, and inflammation, itching and pain, which are diseases based on the activation of TRPV1.
- test substance which reduces the activity of the protein as compared to the control group by measuring the activity of the test substance-treated TRPV1 protein and the test substance-untreated TRPV1 protein as a control group.
- a method for screening a candidate material for preventing or improving wrinkles is provided.
- the present invention provides a screening method using any one or more selected from the group consisting of peptides of SEQ ID NO: 1 to SEQ ID NO: 8 of the present invention.
- the transformant is treated with a TRPV1 specific activator and a TRPV1 activity inhibitor candidate in an experimental group, and the control group is any one selected from the group consisting of the TRPV1 specific activator and a peptide described in SEQ ID NO: 1 to SEQ ID NO: 8. Processing the;
- a method of screening a TRPV1 activity inhibitor comprising comparing the respective measurements of step 3) to select a TRPV1 activity inhibitor candidate exhibiting TRPV1 ion channel activity lower or similar to the control.
- It provides a screening method of the candidate material for preventing skin aging or wrinkle improvement comprising the step of screening the test material showing a TRPV1 ion channel activity lower or similar to the control by comparing the respective measurements of step 3).
- the host cell of step 1) may be a human keratinocyte line, and more preferably, but not limited to, a HaCaT cell line, all cell lines that can be used for ion channel activity studies and high efficiency inhibitor screening. Available.
- the TRPV1 specific active agent of step 2) is preferably UV or capsaicin, and the TRPV1 inhibitory peptide described in SEQ ID NOS: 1 to 8 of the present invention is preferably treated at a concentration of 0.001 to 20 mM. And, it is more preferable to treat with 0.01 to 10 mM, but is not limited thereto.
- Candidates of step 2) may be natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, bacteria or fungi metabolites or bioactive molecules.
- the measurement of TRPV1 ion channel activity of step 3) may be performed by calcium imaging, but is not limited thereto. Any test method capable of confirming the activity of TRPV1 may be used.
- TRPV1 activator Since the peptides described in SEQ ID NO: 1 to SEQ ID NO: 8 of the present invention inhibit TRPV1 activity increased by UV or capsaicin known as TRPV1 activator, and have an effect of improving skin aging and wrinkles based on TRPV1 activity in animal models, Skin aging or anti-wrinkle agents mediated by TRPV1 by selecting TRPV1 activity inhibitors and test substances with similar or higher inhibitory effects compared to the inhibitory effects of peptides set forth in SEQ ID NOS: 1 to 8 on TRPV1 activity Candidates of can be screened.
- the present invention has developed a novel peptide having an anti-aging effect and anti-wrinkle effect.
- the peptide inhibits the expression of MMP and proinflammatory cytokines induced by UV exposure in human keratinocytes and in vivo, and reduces skin thickness and It reduces apoptosis and increases the expression of procollagen reduced by UV exposure, thereby improving skin wrinkles and elasticity due to skin aging and is effective as an agonist of a composition having anti-aging effect.
- FIGS. 1A and 1B are diagrams showing changes in the expression level of MMP-1 induced by UV by each TRPV1 inhibitory peptide.
- 2A to 2H are diagrams showing changes in the amount of proinflammatory cytokine expression induced by UV by each TRPV1 inhibitory peptide.
- FIG. 5 shows increased skin thickness change by UV of hairless mouse skin by TRPV1 inhibitory peptide.
- Fig. 6 is a graph showing the change in the expression level of MMP-13 induced by UV of hairless mouse skin by TRPV1 inhibitory peptide.
- Fig. 7 shows changes in expression levels of MMP-13 and MMP-9 genes induced by UV of hairless mouse skin by TRPV1 inhibitory peptides.
- FIG. 8 is a graph showing changes in the expression level of procollagen genes reduced by UV of hairless mouse skin by TRPV1 inhibitory peptides.
- FIG. 9 is a diagram showing the change of cell thickness increased by UV of hairless mouse skin by TRPV1 inhibitory peptide.
- FIG. 10 shows changes in apoptosis increased by UV in hairless mouse skin by TRPV1 inhibitory peptides.
- novel peptide 1 QRRPSLKSL (SEQ ID NO: 1)
- peptide 2 QRAITILDT (SEQ ID NO: 2)
- peptide 3 RRPSL (SEQ ID NO: 3)
- peptide 4 RAITI (SEQ ID NO: 4)
- peptide 5 MHRQETVDC ( SEQ ID NO: 5)
- Peptide 6 LKKFNARRKL (SEQ ID NO: 6)
- Peptide 7 RQETV (SEQ ID NO: 7)
- Peptide 8 KFNAR (SEQ ID NO: 8).
- HaCaT an immortalized human keratinocyte line
- DMEM Dulbecco's modified Eagle's media
- penicillin 50 mg / mL streptomycin
- FBS phosphate buffered saline
- the HaCaT cells showed emission spectra in the range of 275 and 380 nm 30 minutes after each peptide was treated with HaCaT cells.
- 310-315 nm was irradiated using Philips TL 20W / 12 RS fluorescent sunlight.
- AKodacel filter (TA401 / 407; Kodak) having a wavelength of 290 nm or less was used to prevent UVC, and UV intensity was measured using a Waldmann UV meter.
- UV irradiation the culture medium of the cells was replaced with fresh medium without FBS, and after further incubation, each peptide prepared in ⁇ Example 1> was added to the medium.
- the UV irradiation group was found to increase the expression level of MMP-1 protein compared to the UV irradiation group, and in the UV irradiation and peptide treatment group, the expression of MMP-1 protein increased by UV was increased as the peptide concentration was increased. The amount was found to be significantly reduced (FIGS. 1A and 1B).
- the primers of Table 1 were used, and PCR was performed under conditions of 2 minutes at 50 ° C, 2 minutes at 95 ° C, and then 15 seconds at 95 ° C of 40 cycles and 1 minute at 60 ° C.
- Data were analyzed using 2-DDCT method, expressed as gene expression multiples by normalizing to the expression level of 36B4 gene, and normalized to the intensity of the 36B4 gene, and then increasing or decreasing the ratio compared to UV irradiation group or control group. It was. Each experiment was performed three times, each of which was repeated at least three times.
- Example ⁇ 3-1> the expression level of the MMP-1 gene was significantly increased in the UV irradiation group compared to the UV irradiation group, and in each UV irradiation and peptide treatment group, The expression level of the MMP-1 gene was shown to decrease depending on the treatment concentration (FIGS. 1A and 1B). Therefore, the TRPV1 inhibitory peptide of the present invention was found to act to reduce the expression of MMP-1 by UV stimulation.
- IL-1 ⁇ In order to confirm the cytokine expression change of the TRPV1 inhibitory protein on the UV-irradiated cells, IL-1 ⁇ , IL- in HaCaT cells treated with each TRPV1 peptide, according to the method of Example ⁇ 3-1>. 6, IL-8 and TNF- ⁇ protein expression was measured.
- the TRPV1 inhibitory protein was shown to reduce the expression level of the cytokines increased by UV exposure (Figs. 2A-2H).
- HaCaT cells were cultured in a cover glass, and then cultured in a serum-free medium to which 4 mM Fluo-4 AM (Molecular Probes) was added at room temperature for 45 minutes. After incubation, the cells were washed three times with serum-free medium, and then the cells on the cover glass were transferred to a customized observation chamber and left for 20 minutes. Subsequently, only capsaicin is treated to induce the activity of TRPV1, or the peptides of the present invention are combined with capsaicin to Tyrode's buffer (140 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 2 mM CaCl 2 , 10 mM glucose, respectively).
- Tyrode's buffer 140 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 2 mM CaCl 2 , 10 mM glucose, respectively.
- the TRPV1 inhibitory peptide treated group was found to have a significantly lower Ca 2+ intensity than the capsaicin-only peptide untreated group. Therefore, it was confirmed that the TRPV1 inhibitory peptides of the present invention inhibit the influx of Ca 2+ into keratinocytes (FIG. 3).
- Irradiation intensity at the skin surface was measured using a UV meter (model 585100; Waldmann Co., Villingen-Schwenningen, Germany) and the irradiation intensity at 30 cm from the light source was 1.0 dl / cm 2.
- the minimum erythema dose (MED) measured on the dorsal skin of the mouse for the first time was determined as the minimum irradiation dose required to form a sharp borderline erythema after 24 hours.
- the Skh-1 mice were divided into six groups: (1) UV untreated and carrier treated group, (2) UV untreated and 1 mM peptide treated group, (3) UV irradiated and carrier treated group, (4 ) UV irradiation and 0.01 mM peptide treatment group, (5) 0.1 mM peptide treatment group, (6) UV irradiation and 1 mM peptide treatment group.
- the carrier consisted of ethanol (30%) and polyethylene glycol (70%) and the carrier and peptides were treated on the dorsal skin surface of mice 0 and 24 hours after UV irradiation. The mice were then killed 48 hours after UV irradiation and the skin specimens were biopsied (FIG. 4).
- Subcutaneous fat thickness was measured using a caliper (PEACOCK, Ozaki MFG Co. Ltd., Tokyo, Japan) 24 hours before and 24 hours after UV irradiation of Example ⁇ 5-2>. Specifically, the centerline skin was pinched upward at the neck and the lower part of the tail, and the subcutaneous fat thickness was measured midway between the neck and hips.
- PEACOCK Ozaki MFG Co. Ltd., Tokyo, Japan
- the thickness of the subcutaneous fat was rapidly increased by irradiation with UV, and the thickness of the subcutaneous fat was decreased when the 1 mM TRPV1 inhibitory peptide was treated (FIG. 5).
- the mouse skin tissue of Example ⁇ 5-2> was cold lysis buffer [50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM ethylenediamine tetraacetic acid to which fresh protease inhibitor cocktail (Roche, Indianapolis, IN) was added. (ethylenediamine tetraacetic acid; EDTA), 5 mM phenylmethanesulfonyl fluoride (PMSF), and 1 mM dithiothreitol (DTT), 1% Triton X-100]. The homogenate was then centrifuged at 15,000 g for 30 minutes at 4 ° C. to obtain a supernatant and then stored at ⁇ 70 ° C.
- EDTA ethylenediamine tetraacetic acid
- PMSF phenylmethanesulfonyl fluoride
- DTT dithiothreitol
- Protein content in the lysate was determined by Bradford analysis. Equal amounts of protein were separated on 8-16% Tris-glycine SDS-PAGE gels and electrophoresed onto PVDF membranes. The blots were then blocked for 1 hour at room temperature with blocking buffer and then incubated with monoclonal anti-MMP-13 antibody (Neomarkers, Fremont, CA). As a control, ⁇ -actin levels were measured in the same cell lysates using ⁇ -actin antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). Signal strength was quantified using a densitometer program.
- the expression level of MMP-13 protein was increased in comparison with the UV irradiating group, and in the UV irradiation and peptide treatment groups, the expression level of MMP-13 protein was significantly decreased as the peptide concentration was increased. (FIG. 6).
- RNA was isolated from mouse skin tissue of Example ⁇ 5-2> to synthesize cDNA.
- PCR was performed on a 7500 Real-time PCR System (Applied Biosystems, Foster City, CA) using SYBR_Premix Ex TaqTM (Takara Bio Inc., Shiga, Japan) for quantification of mRNA expression for MMP.
- the primers of Table 2 were used, and PCR was carried out under conditions of 2 minutes at 50 ° C, 2 minutes at 95 ° C, and then 15 seconds at 95 ° C of 40 cycles and 1 minute at 60 ° C.
- the data were analyzed using 2-DDCT method, and normalized to the expression level of 37B4 gene and expressed as a fold of gene expression, and after normalizing to the intensity of the 37B4 gene, yielding an increased or decreased ratio compared to the UV irradiation group or the control group. It was. Each experiment was performed three times, each of which was repeated at least three times.
- the TRPV1 inhibitory peptide was shown to significantly reduce the expression of MMP-13 and MMP-9 genes increased by UV irradiation (FIG. 7). In addition, it was confirmed that expression of the procollagen gene reduced by UV irradiation was significantly increased by TRPV1 inhibitory peptide treatment (FIG. 8).
- hematoxylin and eosin (H & E) staining was performed on the skin tissue of the mouse of Example ⁇ 5-2>. Specifically, mouse skin tissue was fixed in 10% buffered formalin for 24 hours and then embedded in paraffin. Continuous sections (4 ⁇ m) were mounted on silane coated slides, stained with hematoxylin solution for nuclear staining and eosin solution for cytoplasmic staining according to the general method presented in previous studies. Epithelial thickness was then measured using an image analysis program (BMI plus software, BumMi Universe Co., Kyungki, Korea).
- TUNEL staining was performed on mouse skin tissues. Specifically, mouse skin tissue was fixed in 10% buffered formalin for 24 hours and then embedded in paraffin. Consecutive sections (4 ⁇ m) were mounted on silane coated slides and a typical TUNEL staining method using the ApopTagPlus Peroxidase In Situ Apoptosis Kit /S7101MAN.pdf (Millipore TM )] was confirmed apoptosis according to.
- the flexible purified water containing the TRPV1 inhibitory peptide of the present invention as an active ingredient was prepared as shown in Table 3 below.
- Nutritional cream containing the TRPV1 inhibitory peptide of the present invention as an active ingredient was prepared as shown in Table 4 below.
- tablets were prepared by tableting according to a conventional method for producing tablets.
- the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.
- the solution was filled into a 5 ml Type I ampoule made of clear glass, encapsulated under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution.
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Abstract
Description
유전자 | 프라이머 | 서열(5'-3') | 서열번호 |
36B4 | 정방향 프라이머 | TGGGCTCCAAGCAGATGC | 9 |
역방향 프라이머 | GGCTTCGCTGGCTCCCAC | 10 | |
MMP-1 | 정방향 프라이머 | ATTCTACTGATATCGGGGCTTTGA | 11 |
역방향 프라이머 | ATGTCCTTGGGGTATCCGTGTAG | 12 |
유전자 | 프라이머 | 서열(5'-3') | 서열번호 |
마우스 36B4 | 정방향 프라이머 | TGGGCTCCAAGCAGATGC | 9 |
역방향 프라이머 | GGCTTCGCTGGCTCCCAC | 10 | |
마우스 MMP-13 | 정방향 프라이머 | CATCCATCCCGTGACCTTAT | 13 |
역방향 프라이머 | GCATGACTCTCACAATGCGA | 14 | |
마우스 MMP-9 | 정방향 프라이머 | TTGAGTCCGGCAGACAATCC | 15 |
역방향 프라이머 | CCTTATCCACGCGAATGACG | 16 | |
마우스 프로콜라겐 | 정방향 프라이머 | TCGTGACCGTGACCTTGCG | 17 |
역방향 프라이머 | GAGGCACAGACGGCTGAGTAG | 18 |
원료 | 함량(중량부) |
<실시예 1>의 펩티드 1 | 10.00 |
1,3-부틸렌글리콜 | 1.00 |
디소듐이디티에이 | 0.05 |
알란토인 | 0.10 |
디포타슘글리시리제이트 | 0.05 |
시트릭애씨드 | 0.01 |
소듐시트레이트 | 0.02 |
글리세레스-26 | 1.00 |
알부틴 | 2.00 |
하이드로제네이티드캐스터오일 | 1.00 |
에탄올 | 30.00 |
보존제 | 미량 |
착색제 | 미량 |
착향제 | 미량 |
정제수 | 잔량 |
원료 | 함량(중량부) |
<실시예 1>의 펩티드 2 | 10.0 |
1,3-부틸렌 글리콜 | 7.0 |
글리세린 | 1.0 |
D-판테놀 | 0.1 |
식물 추출물 | 3.2 |
마그네슘알루미늄실리케이트 | 0.3 |
PEG-40 스테아레이트 | 1.2 |
스테아릭애씨드 | 2.0 |
폴리소르베이트 60 | 1.5 |
친유형글리세릴스테아레이트 | 2.0 |
소르비탄세스퀴올리에이트 | 1.5 |
세테아릴알코올 | 3.0 |
미네랄오일 | 4.0 |
스쿠알란 | 3.8 |
카르릴릭/카프릭트리글리세라이드 | 2.8 |
식물성 오일 | 1.8 |
디메치콘 | 0.4 |
디포타슘글리시리제이트 | 미량 |
알란토인 | 미량 |
소듐 히아루로네이트 | 미량 |
토코페릴아세테이트 | 적량 |
트리에탄올아민 | 적량 |
보존제 | 적량 |
착향제 | 적량 |
정제수 | 잔량 |
원료 | 함량(중량부) |
세토스테아릴알코올 | 1.6 |
스테아린산 | 1.4 |
친유형모노스테아린산글리세린 | 1.8 |
피이지-100 스테아레이트 | 2.6 |
세스퀴올레인산소르비탈 | 0.6 |
스쿠알렌 | 4.8 |
마카다이아오일 | 2 |
호호바오일 | 2 |
초산토코페롤 | 0.4 |
메칠폴리실록산 | 0.2 |
에칠파라벤 | 0.1 |
초산토코페롤 | 0.4 |
메칠폴리실록산 | 0.2 |
에칠파라벤 | 0.1 |
프로필파라벤 | 0.1 |
1,3-부칠렌글리콜 | 4 |
메칠파라벤 | 0.1 |
산탄검 | 0.1 |
글리세린 | 4 |
d-판데놀 | 0.15 |
알란토인 | 0.1 |
<실시예 1>의 펩티드 3 | 3.5 |
카르보머(2% aq. Sol) | 4 |
트리에탄올아민 | 0.15 |
에탄올 | 3 |
pt 41891 | 0.1 |
p-H20 | 48.3 |
Claims (21)
- 서열번호 1 내지 서열번호 8로 기재되는, TRPV1 활성을 억제하는 펩티드.
- 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 유효성분으로 함유하는 피부노화 방지 또는 주름 개선용 화장료 조성물.
- 청구항 2에 있어서,상기 피부노화는 광노화 또는 자연노화인 화장료 조성물.
- 청구항 3에 있어서,상기 광노화는 UV 노출로 인한 것인 화장료 조성물.
- 청구항 2에 있어서,상기 펩티드는 화장료 조성물에 0.001 내지 20 mM의 농도로 함유되는 화장료 조성물.
- 청구항 2에 있어서,상기 화장료 조성물은 화장수, 에센스, 로션, 크림, 팩, 젤, 연고, 패취 또는 분무제의 제형으로 구성된 군으로부터 선택되는 어느 하나의 제형을 갖는 화장료 조성물.
- 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 유효성분으로 함유하는 피부노화 방지 또는 주름 개선용 약학적 조성물.
- 청구항 7에 있어서,상기 피부노화는 광노화 또는 자연노화인 약학적 조성물.
- 청구항 8에 있어서,상기 광노화는 UV 노출로 인한 것인 약학적 조성물.
- 청구항 7에 있어서,상기 펩티드는 약학적 조성물에 0.001 내지 20 mM의 농도로 함유되는 약학적 조성물.
- 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 유효성분으로 함유하는 피부미백용 약학적 조성물.
- 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 유효성분으로 함유하는 염증, 가려움증, 또는 통증 개선용 약학적 조성물.
- 청구항 11 및 청구항 12 중 어느 한 항에 있어서,상기 펩티드는 약학적 조성물에 0.001 내지 20 mM의 농도로 함유되는 약학적 조성물.
- 약학적으로 유효한 양의 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 개체에 처리 또는 투여하는 단계를 포함하는 피부노화 방지 또는 피부주름 개선 방법.
- 약학적으로 유효한 양의 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 개체에 처리 또는 투여하는 단계를 포함하는 피부미백 방법.
- 약학적으로 유효한 양의 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나 이상을 개체에 처리 또는 투여하는 단계를 포함하는 염증, 가려움증, 또는 통증 개선 방법.
- 1) TRPV1 단백질에 피검물질을 처리하는 단계; 및2) 실험군으로서 상기 피검물질이 처리된 TRPV1 단백질과 대조군으로서 상기 피검물질이 처리되지 않은 TRPV1 단백질의 활성을 측정하여 대조군에 비해 단백질의 활성을 감소시키는 피검물질을 선별하는 단계를 포함하는, 피부노화 방지 또는 주름 개선용 후보물질의 스크리닝 방법.
- 1) TRPV1을 암호화하는 폴리뉴클레오티드를 포함하는 플라스미드가 숙주세포에 형질도입된 형질전환체를 제조하는 단계;2) 상기 형질전환체에 실험군으로 TRPV1 특이적 활성제와 TRPV1 활성 억제제 후보물질을 처리하고, 대조군으로 상기 TRPV1 특이적 활성제와 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나를 처리하는 단계;3) 단계 2)의 상기 실험군과 대조군의 TRPV1 이온 채널 활성을 각각 측정하는 단계; 및,4) 단계 3)의 각각의 측정치를 비교하여 대조군보다 낮거나 유사한 TRPV1 이온 채널 활성을 나타내는 TRPV1 활성 억제제 후보물질을 선별하는 단계를 포함하는 TRPV1 활성 억제제 스크리닝 방법.
- 1) TRPV1을 암호화하는 폴리뉴클레오티드를 포함하는 플라스미드가 숙주세포에 형질도입된 형질전환체를 제조하는 단계;2) 상기 형질전환체에 실험군으로 TRPV1 특이적 활성제와 피검물질을 처리하고, 대조군으로 상기 TRPV1 특이적 활성제와 서열번호 1 내지 서열번호 8로 기재되는 펩티드로 구성된 군으로부터 선택되는 어느 하나를 처리하는 단계;3) 단계 2)의 실험군과 대조군의 TRPV1 이온 채널 활성을 각각 측정하는 단계; 및,4) 단계 3)의 각각의 측정치를 비교하여 대조군보다 낮거나 유사한 TRPV1 이온 채널 활성을 나타내는 피검물질을 선별하는 단계를 포함하는 피부노화 방지 또는 주름 개선용 후보물질의 스크리닝 방법.
- 청구항 18 및 청구항 19 중 어느 한 항에 있어서,상기 TRPV1 특이적 활성제는 UV 또는 캡사이신인 스크리닝 방법.
- 청구항 18 및 청구항 19 중 어느 한 항에 있어서,상기 TRPV1 이온 채널 활성의 측정은 칼슘 이미지화에 의해 수행되는 스크리닝 방법.
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PCT/KR2012/001932 WO2013137505A1 (ko) | 2012-03-16 | 2012-03-16 | 신규한 trpv1 억제 펩티드 및 이를 함유하는 피부노화 방지 또는 주름 개선용 조성물 |
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