WO2017014120A1

WO2017014120A1 - Composition for stimulating trpv1

Info

Publication number: WO2017014120A1
Application number: PCT/JP2016/070639
Authority: WO
Inventors: 典正金川; 伸哉富貴澤; 裕史好田
Original assignee: サントリーホールディングス株式会社
Priority date: 2015-07-17
Filing date: 2016-07-13
Publication date: 2017-01-26
Also published as: JPWO2017014120A1; JP6666912B2; TW201716080A

Abstract

Provided are a composition for stimulating TRPV1, a use for the composition for stimulating TRPV1, and a method for stimulating TRPV1. It was discovered that a specific cyclic dipeptide or a salt thereof has a TRPV1 stimulation effect. The present invention provides an effective and novel means that contributes to the enhancement of various physiological activities such as the acceleration of energy consumption.

Description

TRPV1 stimulating composition

The present invention relates to a composition for stimulating TRPV1. More specifically, the present invention relates to a TRPV1 stimulating composition comprising a cyclic dipeptide or a salt thereof as an active ingredient, the use of the cyclic dipeptide or a salt thereof for stimulating TRPV1, and a method for stimulating TRPV1.

TRPV1 (Transient Receptor Potency Channel, subfamily V, member 1) is a molecule that belongs to the TRP (transient receptor potentio) ion channel superfamily and is known as a receptor involved in the recognition of nociceptive pain and pain. TRPV1 is expressed in sensory nerves and brain, and stimulation of TRPV1 at sensory nerve endings causes depolarization accompanying cation influx and is accepted as a noxious stimulus. As agonists of TRPV1, capsaicin (non-patent document 1), capsiate (non-patent document 2), pepper component (non-patent document 3), pepper component (gingerol and gingerol, etc.) that are pepper components are listed. It is confirmed that it is also activated by heat (threshold: 43 ° C.) and protons.

The biological effects of TRPV1 stimulation include energy consumption promotion. For example, in Non-Patent Document 4, as a result of analysis of body heat production using a normal mouse and a TRPV1 knockout mouse, energy consumption increases when long-term administration of a capsiate, which is a TRPV1 stimulant, to normal mice. It has been demonstrated that the energy consumption increasing effect of capsiate administration is via TRPV1 in the digestive tract.

Regarding TRPV1 stimulation, in Non-Patent Document 5, normal mice and TRPV1 knockout mice were used to examine the effects of capsiates on weight gain and organ fat accumulation, and the effects were observed only in normal mice. From these results, it has been demonstrated that the effects of capsiates on body weight gain and body fat accumulation are mediated by TRPV1 stimulation.

Furthermore, TRPV1 stimulation has been reported to affect muscle and optic nerve disorders. For example, according to Patent Document 1, control of intracellular calcium concentration via TRPV1 is important for activation of the protein synthesis pathway by mTOR and subsequent muscle hypertrophy, and promotes muscle hypertrophy and stimulates muscle atrophy by TRPV1 stimulation. The potential for mitigation is shown. Patent Document 2 suggests that a TRPV1-specific agonist is useful as a preventive or therapeutic agent for optic neuropathy, particularly optic neuropathy caused by glaucoma or glaucomatous visual field stenosis.

Examples of substances that stimulate TRPV1 include polyphenol analogues or hop water extract (Patent Document 3), inhibitor cysteine knot (ICK) peptide (vanillotoxin) (Patent Document 4) and the like in addition to the above agonists.

International Publication No. 2013/141202 JP 2010-24219 A JP 2014-117239 A Japanese translation of PCT publication 2010-510227

An object of the present invention is to provide a composition for stimulating TRPV1. Moreover, the subject of this invention is providing the use of the said composition for stimulating TRPV1, the method etc. which stimulate TRPV1.

As a result of intensive studies on the above problems, the present inventors have found that a specific cyclic dipeptide or a salt thereof has a remarkable TRPV1 stimulating action. Based on this finding, the present inventors have completed the present invention.

That is, the present invention relates to the following, but is not limited thereto.
(1) A TRPV1 stimulating composition comprising a cyclic dipeptide having an amino acid as a structural unit or a salt thereof as an active ingredient,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl Emissions [Cyclo (Asp-Gly)] one selected from the group consisting of or is intended to include two or more, the TRPV1 stimulating composition.
(2) TRPV1 stimulation according to (1) used for energy consumption promotion, body heat production promotion, metabolism promotion, weight gain suppression, organ fat accumulation suppression, muscle increase, muscle atrophy reduction, or optic nerve disorder prevention or treatment Composition.
(3) The composition for stimulating TRPV1 according to (1) or (2), wherein the cyclic dipeptide or a salt thereof is obtained from an animal or plant-derived peptide.
(4) The composition for stimulating TRPV1 according to any one of (1) to (3), which is labeled with a function exhibited by TRPV1 stimulation.
(5) Function indications are “prevent obesity”, “improve obesity”, “suppress body weight gain”, “suppress body fat accumulation”, “suppress visceral fat accumulation” , “Enhance energy consumption”, “enhance body heat production”, “promote metabolism”, “enhance muscle strength”, “suppress muscle weakness”, “improve optic neuropathy”, and “optic neuropathy The composition for stimulating TRPV1 according to (4), which is selected from the group consisting of “prevent”.
(6) The composition for stimulating TRPV1 according to any one of (1) to (5), wherein the composition is an agent.
(7) Use of a cyclic dipeptide having an amino acid as a structural unit or a salt thereof for stimulating TRPV1,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl It is intended to include one or more selected from the group consisting of down [Cyclo (Asp-Gly)], the use.
(8) A method of stimulating TRPV1 using a cyclic dipeptide having an amino acid as a structural unit or a salt thereof as an active ingredient,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl It is intended to include one or more selected from the group consisting of down [Cyclo (Asp-Gly)], the method.

According to the present invention, a composition having an excellent TRPV1 stimulating effect can be provided. By using the composition for stimulating TRPV1 of the present invention in any administration method, effects such as promotion of energy consumption, promotion of body heat production, promotion of metabolism, suppression of weight gain, suppression of organ fat accumulation, etc. via TRPV1 stimulation are obtained. Can do. Moreover, if the composition for stimulating TRPV1 according to the present invention is used, expression of various physiological effects such as the effect of increasing muscles and the effect of suppressing optic nerve damage that have been reported for TRPV1 stimulation can be expected.

1. TRPV1 and TRPV1 stimulation In this specification, “TRPV1” means a molecule that is involved in recognition of nociception in vivo and is identified by the name of Transient Receptor Potential Cation Channel, subfamily V, member 1. TRPV1 is a molecule belonging to the TRP ion channel superfamily, but is distinguished from other members of the family (TRPV2, TRPV3, TRPV4, TRPM2, TRPM4, TRPM5, TRPM8, TRPA1).

As used herein, “TRPV1 stimulation” refers to stimulating TRPV1 to activate it. Various physiological actions can be caused by activating TRPV1. When TRPV1 is activated, it becomes possible for cations to pass therethrough, and this can be used to evaluate TRPV1 stimulation. For example, it is possible to measure and evaluate changes in intracellular calcium ion concentration when a test substance is added to cells expressing TRPV1. Normally, it can be determined that TRPV1 has been stimulated when a change in the concentration is observed (particularly when an increase in intracellular calcium ion concentration is observed). In addition, the degree (strength) of TRPV1 stimulation can be evaluated using, for example, an agonist capsaicin or the like as a control and using the relative value or the like.

2. Cyclic dipeptide In this specification, “cyclic dipeptide” refers to a cyclic dipeptide having a diketopiperazine structure formed by dehydration condensation of an amino group and a carboxyl group of an amino acid. Say. Therefore, the cyclic dipeptide is distinguished from the chain dipeptide. In addition, in this specification, cyclic dipeptide or its salt may be collectively called a cyclic dipeptide. Further, in this specification, as long as the cyclic dipeptides have the same amino acid configuration, any order thereof may be used, for example, [Cyclo (Met-Arg)] and [Cyclo (Arg-Met)] and Represent the same cyclic dipeptide.

In cyclic dipeptides, the terminal portions of two amino acids are linked via an amide bond (that is, the cyclic dipeptide has a cyclic structure formed by the amide bond between the amino terminus and the carboxy terminus. Therefore, cyclic dipeptides are more lipophilic than linear dipeptides with polar carboxyl groups or amino groups exposed at the molecular end (particularly linear dipeptides of the same amino acid composition). It has the characteristics. Therefore, cyclic dipeptides are superior in gastrointestinal permeability and membrane permeability compared to linear dipeptides. This is also clear from the results of compound permeation tests using rat inverted intestinal tracts reported in the past (J. Pharmacol, 1998, 50: 167-172). Cyclic dipeptides are also considered to have increased resistance to various peptidases due to their specific structure.

Cyclic dipeptide or a salt thereof contained as an active ingredient in the present invention is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo ( Leu-Trp)], cycloglycyltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamyl glutamic acid (Cyclo (Glu-Glu)], cycloalanylalanine [Cyclo (Ala-Ala)], cyclomethionylproline [Cyclo (Met-Pro)], cycloprolyl tyrosine [Cyclo (Pro-Tyr)], cycloserylserine [ Cyclo (Ser-Ser)], cycloalanylproline (Cyclo (Ala-Pro)), cycloprolylvaline (Cyclo (Pro-Val)), cycloalanylserine (Cyclo (Ala-Ser)), cycloprolyl Threonine (Cyclo ( Pro-Thr)], and cycloaspartylglycine [Cyclo (Asp-Gly)]. Although the number of cyclic dipeptide or its salt is not specifically limited, In this invention, it is preferable to use 3 or more selected from the cyclic dipeptide mentioned above or its salt as an active ingredient. Among the cyclic dipeptides or salts thereof, cycloglycyltryptophan [Cyclo (Gly-Trp)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucil tryptophan [Cyclo (Leu-Trp)] ], One or more selected from the group consisting of cycloaspartylphenylalanine [Cyclo (Asp-Phe)] and cycloseryltyrosine [Cyclo (Ser-Tyr)], and cycloglycyltryptophan [Cyclo ( Gly-Trp)], cyclohistidylphenylalanine [Cyclo (His-Phe)], and cycloleucyltryptophan [Cyclo (Leu-Trp)] are more preferably one or more.

As used herein, the term “cyclic dipeptide salt” refers to any pharmacologically acceptable salt (including inorganic salts and organic salts) of the cyclic dipeptide, such as sodium salt and potassium salt of the cyclic dipeptide. , Calcium salt, magnesium salt, ammonium salt, hydrochloride, sulfate, nitrate, phosphate, organic acid salt (acetate, citrate, maleate, malate, oxalate, lactate, succinate , Fumarate, propionate, formate, benzoate, picrate, benzenesulfonate, trifluoroacetate, and the like), but are not limited thereto. Cyclic dipeptide salts can be readily prepared by those skilled in the art by any method known in the art.

The cyclic dipeptide used in the present invention can be prepared according to a method known in the art. For example, it may be produced by a chemical synthesis method, an enzymatic method, or a microbial fermentation method, or may be synthesized by dehydration and cyclization of a linear peptide. JP 2003-252896 A, Journal of Peptide や Science, 10, 737-737, 2004. For example, an animal and plant derived peptide heat-treated product rich in cyclic dipeptide can be obtained by further heat-treating an animal and plant derived peptide obtained by subjecting a raw material containing animal and plant derived protein to enzyme treatment or heat treatment. From these points, the cyclic dipeptide or salt thereof used in the present invention may be chemically or biologically synthesized, or may be obtained from an animal or plant derived peptide.

3. Animal and Plant Derived Peptide “Animal and Plant Derived Peptide” in the present specification is not particularly limited. For example, soybean peptide, tea peptide, malt peptide, milk peptide, placenta peptide, collagen peptide and the like can be used. Of these, collagen peptides and soybean peptides are preferred in the present invention. The animal and plant-derived peptide may be a protein prepared from an animal or plant-derived protein or a raw material containing protein, or a commercially available product may be used.

3-1. Soybean peptide As used herein, “soybean peptide” refers to a low molecular weight peptide obtained by subjecting soy protein to enzyme treatment or heat treatment to lower the molecular weight of the protein. Soybeans (scientific name: Glycine max) used as a raw material can be used without restriction of varieties and production areas, and can also be used in processed products such as pulverized products.

3-2. Tea peptide As used herein, “tea peptide” refers to a low molecular weight peptide derived from tea obtained by subjecting a tea (including tea leaves or tea husk) extract to enzyme treatment or heat treatment to lower the protein. . As a tea leaf used as an extraction raw material, a tea leaf (scientific name: Camellia sinensis) manufactured tea leaf leaf, stem, etc. that can be extracted and used can be used. Also, the form is not limited to large leaves or powders. The harvest time of tea leaves can also be selected appropriately according to the desired flavor.

3-3. Malt peptide As used herein, the term “malt peptide” refers to a malt-derived low molecular weight peptide obtained by subjecting an extract obtained from malt or a pulverized product thereof to enzymatic treatment or heat treatment to lower the molecular weight of the protein. . Although the malt peptide used as a raw material can be used without restriction of varieties and production areas, barley malt obtained by germinating barley seeds is particularly preferably used. In the present specification, barley malt may be simply referred to as malt.

3-4. Milk peptide As used herein, “milk peptide” is a product obtained by decomposing milk protein, which is a component derived from natural milk, into a molecule in which at least several amino acids are bound. More specifically, it is obtained by hydrolyzing milk protein such as whey (whey protein) or casein with an enzyme such as proteinase, and filtering and sterilizing and / or concentrating and drying the filtrate. Examples include whey peptides and casein peptides.

3-5. The placenta peptide placenta is the placenta of mammals and has been used as a health food, cosmetics, and pharmaceutical material in recent years because of its excellent functionality. In the present specification, “placenta peptide” refers to a placenta that has been solubilized and reduced in molecular weight by enzyme treatment or subcritical treatment. In addition, although different from the original meaning, extracts obtained from plant placenta are used in health foods, cosmetics, etc. as having a physiological effect equivalent to placenta derived from placenta. be called. The “placenta peptide” in the present specification includes those obtained by subjecting plant placenta to enzyme treatment or subcritical treatment, solubilization and low molecular weight.

3-6. Collagen peptide As used herein, the term “collagen peptide” refers to a low molecular peptide obtained by subjecting collagen or a pulverized product thereof to enzymatic treatment or heat treatment to lower the molecular weight of collagen. Collagen is a major protein in animal connective tissue and is the most abundant protein in mammalian bodies including humans.

4). As described above animal and plant derived peptides Cook, by high-temperature heat treatment plants and animals derived peptides, it is possible to obtain plants and animals derived peptides heat treatment comprising a cyclic dipeptide rich. In this specification, “high temperature heat treatment” means that the treatment is performed for a certain period of time at a temperature of 100 ° C. or higher and a pressure exceeding atmospheric pressure. As the high-temperature and high-pressure treatment device, a pressure-resistant extraction device, a pressure cooker, an autoclave, or the like can be used according to conditions.

The temperature in the high-temperature heat treatment is not particularly limited as long as it is 100 ° C or higher, but is preferably 100 ° C to 170 ° C, more preferably 110 ° C to 150 ° C, and still more preferably 120 ° C to 140 ° C. In addition, this temperature shows the value which measured the exit temperature of the extraction column, when using a pressure-resistant extraction apparatus as a heating apparatus, and when using an autoclave as a heating apparatus, it is the temperature of the center temperature in a pressure vessel. The measured value is shown.

The pressure in the high-temperature heat treatment is not particularly limited as long as it is a pressure exceeding atmospheric pressure, but is preferably 0.101 MPa to 0.79 MPa, more preferably 0.101 MPa to 0.60 MPa, and even more preferably 0.101 MPa to 0. 48 MPa.

The high-temperature heat treatment time is not particularly limited as long as a processed product containing a cyclic dipeptide is obtained, but is preferably about 15 minutes to 600 minutes, more preferably about 30 minutes to 500 minutes, and even more preferably about 60 minutes to 300 minutes. It is.

In addition, the high-temperature heat treatment conditions for the animal and plant derived peptides are not particularly limited as long as a processed product containing a cyclic dipeptide is obtained, but preferably [temperature: pressure: time] is [100 ° C. to 170 ° C .: 0.101 MPa to 0.001. 79 MPa: 15 minutes to 600 minutes], more preferably [110 ° C. to 150 ° C .: 0.101 MPa to 0.60 MPa: 30 minutes to 500 minutes], even more preferably [120 ° C. to 140 ° C .: 0.101 MPa to 0 48 MPa: 60 minutes to 300 minutes].

In addition, you may perform processes, such as filtration, centrifugation, concentration, ultrafiltration, lyophilization | freeze-drying, and powdering, with respect to the obtained heat-processed peptide derived from animals and plants. In addition, if the specific cyclic dipeptide in the heat-treated product of animal and plant derived peptides does not satisfy the desired content, the specific cyclic dipeptide that is deficient may be appropriately added using other animal or plant derived peptides, commercial products, or synthetic products. it can.

5). 5. TRPV1 stimulating composition 5-1. Cyclic dipeptide-containing TRPV1 stimulating composition One aspect of the present invention is a TRPV1 stimulating composition comprising a specific cyclic dipeptide or a salt thereof as an active ingredient.

The composition for stimulating TRPV1 of the present invention comprises cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], Cycloglycyltryptophan [Cyclo (Gly-Trp)], cyclophenylalanyltryptophan [Cyclo (Phe-Trp)], cycloseryltyrosine [Cyclo (Ser-Tyr)], cycloglutamylglutamate [Cyclo (Glu-Glu)] , Cycloalanylalanine [Cyclo (Ala-Ala)], cyclomethionylproline [Cyclo (Met-Pro)], cycloprolyl tyrosine [Cyclo (Pro-Tyr)], cycloserylserine [Cyclo (Ser-Ser)] ], Cycloalanylproline [Cyclo (Ala-Pro)], cycloprolylvaline [Cyclo (Pro-Val)], cycloalanylserine [Cyclo (Ala-Ser)], cycloprolylthreonine [Cyclo (Pro- Thr)], and cycloaspartyl Is intended to include one or more cyclic dipeptide or a salt thereof is selected from the group consisting of lysine [Cyclo (Asp-Gly)] as an active ingredient. The number of cyclic dipeptides or salts thereof included in the TRPV1 stimulating composition of the present invention is not particularly limited, but in the present invention, three or more selected from the above-mentioned cyclic dipeptides or salts thereof are preferably included. Among the cyclic dipeptides or salts thereof, cycloglycyltryptophan (Cyclo (Gly-Trp)), cyclohistidylphenylalanine (Cyclo (His-Phe)), cycloleusyl tryptophan (Cyclo (Leu-Trp)), One or more selected from the group consisting of cycloaspartylphenylalanine [Cyclo (Asp-Phe)] and cycloseryltyrosine [Cyclo (Ser-Tyr)] are preferred, and cycloglycyltryptophan [Cyclo (Gly- Trp)], cyclohistidylphenylalanine [Cyclo (His-Phe)], and cycloleucyltryptophan [Cyclo (Leu-Trp)] are more preferably one or more.

The content of the cyclic dipeptide or the salt thereof in the TRPV1 stimulating composition of the present invention is not particularly limited as long as the desired effect of the present invention is obtained in consideration of its administration form, administration method and the like. It is not something. For example, when soybean peptide, tea peptide, malt peptide, milk peptide, placenta peptide, or collagen peptide is used as a raw material, the total content of cyclic dipeptide or salt thereof in the composition of the present invention is preferably 200 ppm / Brix or more, preferably Is 300 ppm / Brix or more, 5000 ppm / Brix or less, preferably 4000 ppm / Brix or less, and typically 200 to 5000 ppm / Brix, preferably 300 to 4000 ppm / Brix. In addition, cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)] in the TRPV1 stimulating composition of the present invention. Cycloglycyltryptophan [Cyclo (Gly-Trp)], cyclophenylalanyltryptophan [Cyclo (Phe-Trp)], cycloseryltyrosine [Cyclo (Ser-Tyr)], cycloglutamylglutamate [Cyclo (Glu-Glu) ), Cycloalanylalanine [Cyclo (Ala-Ala)], cyclomethionylproline [Cyclo (Met-Pro)], cycloprolyl tyrosine [Cyclo (Pro-Tyr)], cycloserylserine [Cyclo (Ser-Ser )], Cycloalanylproline [Cyclo (Ala-Pro)], cycloprolylvaline [Cyclo (Pro-Val)], cycloalanylserine [Cyclo (Ala-Ser)], cycloprolylthreonine [Cyclo (Pro -Thr)), cycloasphal The content of luglycine [Cyclo (Asp-Gly)] or the corresponding salt is 1.0 ppm / Brix or more, preferably 3.0 ppm / Brix or more, 3000 ppm / Brix or less, preferably 2000 ppm / Brix. Typically, it is 1.0 to 3000 ppm / Brix, and preferably 3.0 to 2000 ppm / Brix. In this invention, content of cyclic dipeptide or its salt is represented by the quantity per Brix (Brix: Bx) as above-mentioned. In this specification, “amount per Brix” means an amount determined by a value corresponding to a mass percentage of a sucrose solution at 20 ° C. (an aqueous solution containing only sucrose as a solute). Unless otherwise specified, “ppm” used in the present specification means ppm of weight / volume (w / v), and 1.0 ppm / Brix is 0.1 mg / wt when the specific gravity of the solvent is 1. Converted to mL and converted to 0.01% by weight.

The content of the cyclic dipeptide or a salt thereof can be measured according to a known method. For example, it can be measured using LC-MS / MS or a saccharimeter.

Moreover, the TRPV1 stimulating composition of the present invention may contain a heat-treated animal or plant-derived peptide heat-treated product containing one or more of the cyclic dipeptides or salts thereof. Such a heat-treated product derived from animals and plants is not particularly limited, but a heat-treated product of soybean peptide and a heat-treated product of collagen peptide are preferable.

When animal and plant-derived peptide heat-treated products are used, the content in the TRPV1 stimulating composition of the present invention is such that the desired effect of the present invention can be obtained in consideration of its administration form, administration method, etc. Well, not particularly limited. For example, the content is 0.001% by weight or more, preferably 0.01% by weight or more, more preferably 0.1% by weight or more with respect to the total weight of the composition of the present invention. The content of the heat-treated product derived from animals and plants is 99% by weight or less, preferably 50% by weight or less, more preferably 10% by weight or less based on the total weight of the composition of the present invention.

5-2. Other Components The TRPV1 stimulating composition of the present invention can contain any additive and any commonly used component in addition to the above active ingredients, depending on the form. Examples of these additives and / or ingredients include vitamins such as vitamin E and vitamin C, bioactive ingredients such as minerals, nutritional ingredients, and fragrances, as well as excipients and binders incorporated in the formulation. , Emulsifiers, tonicity agents (isotonic agents), buffers, solubilizers, preservatives, stabilizers, antioxidants, colorants, coagulants, or coating agents, but are not limited thereto. It is not something.

5-3. Use The TRPV1 stimulating composition of the present invention is characterized by containing the above-mentioned active ingredient, and the active ingredient stimulates TRPV1 to cause various physiological actions. In the present invention, by stimulating and activating TRPV1, the present invention can be applied to the use of energy consumption promotion, body heat production promotion, metabolism promotion, weight gain suppression, organ fat accumulation suppression, muscle increase, muscle atrophy reduction, optic nerve It is possible to effectively prevent or treat a disorder. Therefore, the composition of the present invention is a composition for use in promoting energy consumption, promoting body heat production, promoting metabolism, inhibiting weight gain, inhibiting organ fat accumulation, increasing muscle mass, reducing muscle atrophy, or preventing optic nerve damage or A therapeutic TRPV1 stimulating composition. Based on these uses, the composition for stimulating TRPV1 of the present invention comprises a composition for promoting energy consumption, a composition for promoting body heat production, a composition for promoting metabolism, a composition for suppressing weight gain, and a composition for suppressing accumulation of organ fat. It can also be a composition for increasing or decreasing muscles, a composition for reducing muscle atrophy, or a composition for preventing or treating optic neuropathy. In this specification, the terms “prevention” and “treatment” include both the concepts of making the current state a better state and preventing the current state from becoming worse than the current state. Thus, terms such as improvement, recovery, mitigation, mitigation can also be included in these.

The TRPV1 stimulating composition of the present invention can be prepared by using, for example, the above-described other components and the like according to a known method, such as solid agents such as tablets, granules, powders, powders, or capsules, normal solutions, suspensions, and the like. It can be formulated into a suspension or a liquid such as an emulsion. These compositions can be taken with water or the like as it is. Moreover, after preparing the form (for example, powder form and granule form) which can be mix | blended easily, it can use, for example as a raw material of a pharmaceutical.

The TRPV1 stimulating composition of the present invention can be provided in the form of an agent as an example, but is not limited to this form. The agent can be provided as a composition as it is or as a composition containing the agent. Examples of the composition of the present invention include, but are not limited to, a pharmaceutical composition, a food / beverage product composition, a food composition, a beverage composition, a cosmetic composition, and the like. Non-limiting examples of food compositions include functional foods, health supplements, functional nutrition foods, special foods, foods for specified health use, dietary supplements, diet foods, health foods, supplements, food additives, etc. Can be mentioned.

The TRPV1 stimulating composition of the present invention can be applied to any therapeutic use (medical use) or non-therapeutic use (non-medical use). Specifically, pharmaceuticals, quasi-drugs, cosmetics, etc. and the Pharmaceutical Affairs Law do not belong to these, but energy consumption promotion, body heat production promotion, metabolism promotion, weight gain suppression, organ fat accumulation suppression, muscle increase, Application to the reduction of muscle atrophy, or use as a composition that explicitly or implicitly appeals for the effect of preventing or treating optic neuropathy, etc.

In another aspect, the present invention relates to the TRPV1 stimulating composition, which is labeled with the function exhibited by TRPV1 stimulation. Such indications or functional indications are not particularly limited, and for example, “prevent obesity”, “improve obesity”, “suppress body weight gain”, “suppress body fat accumulation”, “ Suppress visceral fat accumulation, increase energy consumption, increase body heat production, promote metabolism, prevent obesity, improve obesity, increase muscle strength ”,“ Suppress muscular weakness ”,“ improve optic neuropathy ”,“ prevent optic neuropathy ”and the like, and the description of consent with these is also included in the display. In the present specification, indications such as the indication and the functionality indication may be attached to the composition itself, or may be attached to a container or packaging of the composition.

The TRPV1 stimulating composition of the present invention can be taken by an appropriate method according to the form. The intake method is not particularly limited as long as the cyclic dipeptide or a salt thereof according to the present invention can be transferred into the circulating blood. For example, oral solid preparations such as tablets, coated tablets, granules, powders, or capsules, oral liquid preparations such as oral liquids, syrups, injections, external preparations, suppositories, or transdermal absorption agents, etc. However, the present invention is not limited thereto. In the present specification, “ingestion” is used to include all aspects such as ingestion, taking, or drinking.

The application amount of the composition for stimulating TRPV1 of the present invention is appropriately set according to the form, administration method, purpose of use, and age, weight, and symptom of the patient or animal to be administered, and is not constant. Although the effective human intake of the composition of the present invention is not constant, for example, the total amount of the cyclic dipeptide or salt thereof as the active ingredient is preferably 10 mg or more, more preferably 100 mg per day for a human body weight of 50 kg. That's it. Further, administration may be performed once or several times within one day within a desired dose range. The administration period is also arbitrary. In addition, the effective human intake of the composition of the present invention refers to the intake of the composition for stimulating TRPV1 of the present invention showing an effective effect in humans.

The subject of application of the composition for stimulating TRPV1 of the present invention is preferably a human, but domestic animals such as cattle, horses and goats, pet animals such as dogs, cats and rabbits, or mice, rats, guinea pigs, monkeys, etc. It may be a laboratory animal. When a non-human animal is administered to a subject, the amount used per day for about 20 g per mouse is the content of the active ingredient in the composition, the state of the subject, weight, sex, age, etc. Usually, the total amount of the cyclic dipeptide or its salt is preferably 10 mg / kg or more, more preferably 100 mg / kg or more.

6). Use of a cyclic dipeptide or a salt thereof for stimulating TRPV1 One embodiment of the present invention is the use of a specific cyclic dipeptide or a salt thereof having an amino acid as a constituent unit for stimulating TRPV1. Preferably, cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucil tryptophan [Cyclo (Leu-Trp)], cycloglycyltryptophan [Cyclo ( Gly-Trp)], cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate [Cyclo (Glu-Glu)], cycloalanylalanine [Cyclo (Ala-Ala)], cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), cycloalanyl proline [ Cyclo (Ala-Pro)], cycloprolyl valine [Cyclo (Pro-Val)], cycloalanylserine [Cyclo (Ala-Ser)], cycloprolylthreonine [Cyclo (Pro-Thr)], and cycloaspa Rutileglycine (Cyclo (Asp-Gly)) One selected from the group consisting or more than one cyclic dipeptide or a use for stimulating TRPV1 salt thereof. More preferably, the cyclic dipeptide or a salt thereof containing three or more is used for stimulating TRPV1.

Uses of the present invention include, for example, energy consumption promotion, body heat production promotion, metabolism promotion, weight gain suppression, organ fat accumulation suppression, muscle increase, muscle atrophy reduction, or prevention or treatment of optic neuropathy , Use of the cyclic dipeptide or a salt thereof is included, but is not limited thereto. In addition, the use is a use in a human or non-human animal, and may be a therapeutic use or a non-therapeutic use. Here, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by treatment.

7). Method for Stimulating TRPV1 One embodiment of the present invention is a method for stimulating TRPV1 using a specific cyclic dipeptide having an amino acid as a structural unit or a salt thereof as an active ingredient. The method is preferably cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Tryptophan [Cyclo (Gly-Trp)], cyclophenylalanyltryptophan [Cyclo (Phe-Trp)], cycloseryltyrosine [Cyclo (Ser-Tyr)], cycloglutamylglutamate [Cyclo (Glu-Glu)], cycloaralan Nylalanine [Cyclo (Ala-Ala)], cyclomethionylproline [Cyclo (Met-Pro)], cycloprolyl tyrosine [Cyclo (Pro-Tyr)], cycloserylserine [Cyclo (Ser-Ser)], cyclo Alanylproline [Cyclo (Ala-Pro)], cycloprolylvaline [Cyclo (Pro-Val)], cycloalanylserine [Cyclo (Ala-Ser)], cycloprolylthreonine [Cyclo (Pro-Thr)] And cycloaspartyl glycine [Cycl o (Asp-Gly)] is a method for stimulating TRPV1, comprising using one or more cyclic dipeptides selected from the group consisting of o (Asp-Gly)] or a salt thereof as an active ingredient. More preferably, it is a method of stimulating TRPV1, comprising using as an active ingredient a substance comprising three or more selected from the cyclic dipeptides or salts thereof.

Another aspect related to the method is a method of stimulating TRPV1, comprising administering to a subject in need of TRPV1 stimulation a therapeutically effective amount of a specific cyclic dipeptide or a salt thereof as an active ingredient. Preferably, cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucil tryptophan [Cyclo (Leu-Trp)], cycloglycyltryptophan [Cyclo ( Gly-Trp)], cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate [Cyclo (Glu-Glu)], cycloalanylalanine [Cyclo (Ala-Ala)], cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), cycloalanyl proline [ Cyclo (Ala-Pro)], cycloprolyl valine [Cyclo (Pro-Val)], cycloalanylserine [Cyclo (Ala-Ser)], cycloprolylthreonine [Cyclo (Pro-Thr)], and cycloaspa Rutileglycine (Cyclo (Asp-Gly)) Comprising comprising administering a therapeutically effective amount of a one or more cyclic dipeptide or a salt thereof as an active ingredient is selected from the group, is a method of stimulating the TRPVl. More preferably, it is a method of stimulating TRPV1, comprising administering a therapeutically effective amount using as an active ingredient a substance containing three or more selected from the cyclic dipeptides or salts thereof.

In the above method, the subject requiring TRPV1 stimulation is the same as the subject of application of the TRPV1 stimulation composition of the present invention. In the present specification, the therapeutically effective amount is an amount by which TRPV1 is stimulated when the composition for stimulating TRPV1 of the present invention is administered to the subject as compared with a subject not administered. . The specific effective amount is appropriately set according to the administration form, administration method, purpose of use and age, weight, symptom, etc. of the subject and is not constant.

In the method of the present invention, the specific cyclic dipeptide or a salt thereof may be administered as it is or as a composition containing the specific cyclic dipeptide or a salt thereof so that the therapeutically effective amount is obtained.

According to the method of the present invention, it is possible to stimulate TRPV1 without causing side effects.

Hereinafter, the present invention will be described in more detail with reference to examples, but the scope of the present invention is not limited thereby. Those skilled in the art can use the method of the present invention with various changes and modifications, and these are also included in the scope of the present invention.

Example 1. Examination of TRPV1 stimulating action by cyclic dipeptide These TRPV1 stimulating actions were evaluated using various chemically synthesized cyclic dipeptide preparations. Specifically, CHO cells (Chinese hamster ovary-derived cells) expressing human TRPV1 were suspended in 0.1% FBS-containing DMEM (Invitrogen), and 384-well microarrays at 3.5 × 10 ⁴ cells / 90 μL / well. Plates were seeded. Next, 20 mM Hepes buffer (Invitrogen) (pH 7.4) containing a fluorescent probe (Calcium 4, Molecular Device) was added to each well and allowed to equilibrate at 37 ° C. for 60 minutes and then at 22 ° C. for 15 minutes. Thereafter, the assay plate was placed in a microplate reader (CellLux, PerkinElmer), and the fluctuation (response rate) of intracellular calcium ion concentration due to addition of a cyclic dipeptide solution or a reference agonist solution was measured by fluorescence intensity. As a reference agonist solution, a capsaicin solution prepared so that the final concentration was 1 μM was used.

Regarding the TRPV1 stimulating action of the cyclic dipeptide preparation, the change in the intracellular calcium ion concentration (response rate) when the reference agonist solution is added is defined as 100%, and the response rate when the cyclic dipeptide preparation is added is a relative value. (%) Was calculated and evaluated. The number of tests was n = 2, and the average value of the response rate when the cyclic dipeptide preparation was added was determined. The results are shown in Table 1. The concentration in the table indicates the final concentration of the cyclic dipeptide in the assay system.

From the above results, it was clarified that all of the cyclic dipeptides shown in Table 1 have a TRPV1 stimulating action.

Example 2 Examination of TRPV1 stimulation action of heat-treated collagen peptide and heat-treated soybean peptide (1) Preparation of heat-treated collagen peptide Heat-treated collagen peptide was used as the heat-treated collagen peptide. The collagen peptide heat-treated product was produced by subjecting a collagen peptide to high-temperature and high-pressure treatment in a liquid. Specifically, distilled water was added at a concentration of 10 g / 100 ml to collagen peptide (HACP-50, manufactured by Zerice Co., Ltd.), placed in an autoclave (manufactured by Tommy Seiko Co., Ltd.), 135 ° C, 0.31 MPa, high temperature and high pressure for 10 hours. Processing was added.
(2) Preparation of heat-treated soybean peptide A soybean peptide heat-treated product was used as the soybean peptide heat-treated product. The soybean peptide heat-treated product was produced by treating soybean peptide in a liquid at high temperature and high pressure. Specifically, about 15 ml of distilled water was added to 3 g of soy peptide (Hi-New AM, manufactured by Fuji Oil Co., Ltd.), respectively, and placed in an autoclave (produced by Tommy Seiko Co., Ltd.) at 135 ° C., 0.31 MPa, 3 hours. High temperature and high pressure treatment was added.
(3) Evaluation of TRPV1 stimulating action These TRPV1 stimulating actions were evaluated using freeze-dried products of the heat-treated collagen peptide and the heat-treated soybean peptide prepared as described above. Specifically, CHO cells (Chinese hamster ovary-derived cells) expressing human TRPV1 were suspended in 0.1% FBS-containing DMEM (Invitrogen), and 384-well microarrays at 3.5 × 10 ⁴ cells / 90 μL / well. Plates were seeded. Next, 20 mM Hepes buffer (Invitrogen) (pH 7.4) containing a fluorescent probe (Calcium 4, Molecular Device) was added to each well and allowed to equilibrate at 37 ° C. for 60 minutes and then at 22 ° C. for 15 minutes. Thereafter, the assay plate is placed in a microplate reader (CellLux, PerkinElmer), and a collagen peptide heat-treated product or a soybean peptide heat-treated product or a reference agonist reconstituted so that the final concentration in the assay system is 1.0 mg / mL Variation (response rate) of intracellular calcium ion concentration due to addition of the solution was measured by fluorescence intensity. As a reference agonist solution, a capsaicin solution prepared so that the final concentration was 1 μM was used.

Regarding the TRPV1 stimulating action of the heat-treated collagen peptide or heat-treated soy peptide, the response rate when the test material is added is defined as 100% of the change (response rate) in the intracellular calcium ion concentration when the reference agonist solution is added. The relative value (%) was calculated and evaluated. The number of tests was n = 2, and the average value of the response rate when the test material was added was determined. The results are shown in Table 2.

From the above results, it was revealed that both the collagen peptide heat-treated product and the soybean peptide heat-treated product have a TRPV1 stimulating action. It was considered that the cyclic dipeptides contained in various materials (for example, the cyclic dipeptides shown in Table 1) may contribute as one of the factors to the TRPV1 stimulating effect of these materials.

The present invention provides a composition for stimulating TRPV1 containing a specific cyclic dipeptide or a salt thereof as an active ingredient. Since the present invention provides a new means for enhancing various physiological activities such as promotion of energy consumption, the industrial applicability is high.

Claims

A composition for stimulating TRPV1 containing, as an active ingredient, a cyclic dipeptide having an amino acid as a structural unit or a salt thereof,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl Emissions [Cyclo (Asp-Gly)] one selected from the group consisting of or is intended to include two or more, the TRPV1 stimulating composition.
The composition for stimulating TRPV1 according to claim 1, which is used for energy consumption promotion, body heat production promotion, metabolism promotion, weight gain suppression, organ fat accumulation suppression, muscle increase, muscle atrophy reduction, or optic nerve disorder prevention or treatment. .
The composition for stimulating TRPV1 according to claim 1 or 2, wherein the cyclic dipeptide or a salt thereof is obtained from an animal or plant-derived peptide.
The composition for TRPV1 stimulation according to any one of claims 1 to 3, which is labeled with a function exhibited by TRPV1 stimulation.
Function indications are “prevent obesity”, “improve obesity”, “suppress body weight gain”, “suppress body fat accumulation”, “suppress visceral fat accumulation”, “energy Increase consumption, Increase body heat production, Promote metabolism, Strengthen muscle strength, Suppress muscle weakness, Improve optic neuropathy, and Prevent optic neuropathy The composition for stimulating TRPV1 according to claim 4, which is selected from the group consisting of:
The composition for stimulating TRPV1 according to any one of claims 1 to 5, wherein the composition is an agent.
Use of a cyclic dipeptide having amino acid as a structural unit or a salt thereof for stimulating TRPV1,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl It is intended to include one or more selected from the group consisting of down [Cyclo (Asp-Gly)], the use.
A method of stimulating TRPV1, using a cyclic dipeptide having an amino acid as a structural unit or a salt thereof as an active ingredient,
The cyclic dipeptide or a salt thereof is cycloaspartylphenylalanine [Cyclo (Asp-Phe)], cyclohistidylphenylalanine [Cyclo (His-Phe)], cycloleucyltryptophan [Cyclo (Leu-Trp)], cycloglycyl. Syltryptophan (Cyclo (Gly-Trp)), cyclophenylalanyltryptophan (Cyclo (Phe-Trp)), cycloseryltyrosine (Cyclo (Ser-Tyr)), cycloglutamylglutamate (Cyclo (Glu-Glu)), cyclo Alanylalanine (Cyclo (Ala-Ala)), cyclomethionyl proline (Cyclo (Met-Pro)), cycloprolyl tyrosine (Cyclo (Pro-Tyr)), cycloserylserine (Cyclo (Ser-Ser)), Cycloalanylproline (Cyclo (Ala-Pro)), Cycloprolylvaline (Cyclo (Pro-Val)), Cycloalanylserine (Cyclo (Ala-Ser)), Cycloprolylthreonine (Cyclo (Pro-Thr)) And cycloaspartyl It is intended to include one or more selected from the group consisting of down [Cyclo (Asp-Gly)], the method.