Vasodilator and use thereof

Abstract

To provide a novel use of a collagen-derived tripeptide or a collagen-derived dipeptide.SOLUTION: A vasodilator comprises a collagen-derived tripeptide and / or a collagen-derived dipeptide.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a vasodilator and its use. [Background technology]

[0002] In recent years, it has been known that compositions containing tripeptides obtained by hydrolyzing collagen have effects such as promoting collagen production, promoting hyaluronic acid production, and promoting skin collagen synthesis, and are useful for improving skin beauty (Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2012-501320 Summary of the Invention [Problem to be solved by the invention]

[0004] Collagen is a major protein that constitutes the connective tissues of animals, and there is a demand for further utilization of collagen.

[0005] Therefore, an object of the present invention is to provide a new use of collagen-derived tripeptides or collagen-derived dipeptides. [Means for solving the problem]

[0006] In order to achieve the above object, the vasodilator of the present invention contains at least one of a collagen-derived tripeptide and a collagen-derived dipeptide.

[0007] The therapeutic agent for periodontal disease of the present invention comprises the vasodilator of the present invention.

[0008] The oral composition of the present invention contains the vasodilator of the present invention.

[0009] The food composition of the present invention contains the vasodilator of the present invention. Effect of the Invention

[0010] According to the present invention, blood vessels can be dilated. [Brief description of the drawings]

[0011]

Figure 1

Figure 2

Figure 3

Figure 4

[0012] <Vasodilator> As described above, the vasodilator of the present invention contains at least one of a collagen-derived tripeptide (hereinafter also referred to as "tripeptide") and a collagen-derived dipeptide (hereinafter also referred to as "dipeptide"). The vasodilator of the present invention is characterized in that it contains at least one of a collagen-derived tripeptide and a collagen-derived dipeptide, and other configurations and conditions are not particularly limited. The vasodilator of the present invention can dilate blood vessels.

[0013] As a result of intensive research, the present inventors discovered that collagen-derived tripeptides and collagen-derived dipeptides dilate gingival blood vessels when administered to the oral mucosa, and thus arrived at the present invention. This was the first discovery by the present inventors. According to the present invention, the blood vessels of the subject can be dilated, and therefore, for example, blood flow of the subject can be improved. Therefore, the vasodilator of the present invention can also be called, for example, a blood flow improver.

[0014] In the present invention, "vasodilation" means, for example, that the blood vessels of a subject are dilated. Specifically, "vasodilation" means that the blood vessels of a subject are significantly dilated compared to when the vasodilator of the present invention is not administered. The "blood vessels of a subject" include, for example, blood vessels in the gums. The blood vessels of a subject are, for example, blood vessels with a small blood vessel diameter. The "small blood vessels" are, for example, blood vessels with a blood vessel diameter of 10 μm or less. The blood vessel diameter means, for example, the inner diameter of a blood vessel. The time when the blood vessel diameter is measured is, for example, the late vasoconstriction phase (corresponding to the late diastole phase of the heart). The blood vessel diameter can be measured, for example, as the longest diameter of the area where blood (red blood cells) are present when the blood vessels of a subject are optically observed.

[0015] Collagen protein is composed of a polypeptide in which the amino acid sequence Gly-XX (X is any amino acid residue) is repeated multiple times as a constituent unit. Therefore, the collagen-derived tripeptide is a peptide composed of any three consecutive amino acids in collagen protein, including a tripeptide composed of glycine and any two amino acids, and the any amino acid is an amino acid selected from the group consisting of proline, hydroxyproline, glutamic acid, alanine, and arginine. Specific examples of the collagen-derived tripeptide are not particularly limited, and include tripeptides composed of glycine, proline, and hydroxyproline, and tripeptides composed of alanine, hydroxyproline, and glycine. In each tripeptide, the order of the amino acid sequence is not particularly limited and can be any order. When the tripeptide is a tripeptide composed of glycine, proline, and hydroxyproline, the tripeptide is, for example, glycine-proline-hydroxyproline or proline-hydroxyproline-glycine from the N-terminus. When the tripeptide is a tripeptide composed of alanine, hydroxyproline, and glycine, the collagen-derived tripeptide is, for example, alanine-hydroxyproline-glycine from the N-terminus. The tripeptide is preferably glycine-proline-hydroxyproline. The collagen-derived tripeptide may be composed of one type of tripeptide or may be composed of multiple types of tripeptides.

[0016] The collagen-derived dipeptide is a peptide composed of any two consecutive amino acids in a collagen protein, and includes at least two amino acids selected from, for example, glycine, proline, hydroxyproline, glutamic acid, alanine, and arginine. Specific examples of the dipeptide include, but are not limited to, a dipeptide composed of proline and hydroxyproline, a dipeptide composed of alanine and hydroxyproline, a dipeptide composed of leucine and hydroxyproline, a dipeptide composed of isoleucine and hydroxyproline, a dipeptide composed of phenylalanine and hydroxyproline, a dipeptide composed of glycine and proline, a dipeptide composed of glycine and hydroxyproline, a dipeptide composed of glycine and glutamic acid, a dipeptide composed of glycine and alanine, a dipeptide composed of glycine and arginine, a dipeptide composed of proline and glutamic acid, a dipeptide composed of proline and alanine, a dipeptide composed of proline and arginine, a dipeptide composed of hydroxyproline and glutamic acid, a dipeptide composed of hydroxyproline and arginine, a dipeptide composed of glutamic acid and alanine, a dipeptide composed of glutamic acid and arginine, and a dipeptide composed of alanine and arginine. When the dipeptide is a dipeptide composed of proline and hydroxyproline, the dipeptide is, for example, proline-hydroxyproline from the N-terminus.When the dipeptide is a dipeptide composed of alanine and hydroxyproline, the dipeptide is, for example, alanine-hydroxyproline from the N-terminus.When the dipeptide is a dipeptide composed of leucine and hydroxyproline, the dipeptide is, for example, leucine-hydroxyproline from the N-terminus.When the dipeptide is a dipeptide composed of isoleucine and hydroxyproline, the collagen-derived dipeptide is, for example, isoleucine-hydroxyproline from the N-terminus. When the dipeptide is a dipeptide composed of phenylalanine and hydroxyproline, the dipeptide is, for example, phenylalanine-hydroxyproline from the N-terminus. The dipeptide is preferably proline-hydroxyproline. The collagen-derived dipeptide may be composed of one type of dipeptide or multiple types of dipeptides.

[0017] In the tripeptides and dipeptides, the hydroxyproline is not particularly limited and may be, for example, 3-Hyp (3-hydroxyproline) or 4-Hyp (4-hydroxyproline).

[0018] In the present invention, the tripeptides and dipeptides may be modified. That is, the amino acid residues constituting the tripeptides and dipeptides may be natural amino acid residues or modified amino acid residues (non-natural amino acid residues). Therefore, in the present invention, one, a plurality, or all of the amino acid residues constituting the tripeptides and dipeptides are natural amino acid residues or modified amino acid residues. The natural amino acid residues are alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, pyrrolysine, proline, glutamine, arginine, serine, threonine, selenocysteine, valine, tryptophan, or tyrosine.

[0019] When the tripeptide and dipeptide are modified, each peptide can be called a modified peptide. The modified peptide is, for example, a peptide in which some or all of the amino acids constituting the tripeptide or dipeptide are modified. The modification may be, for example, a modification of the amino acids constituting the tripeptide or dipeptide, or a modification of the peptide. Specific examples of the modification include phosphorylation of serine or threonine; sulfation of tyrosine; modification of amino groups such as biotinylation, myristoylation, palmitoylation, acetylation, maleimidation, malonylation, and methylation (e.g., Lys(Me), Lys(Me)2, Lys(Me)3, Arg(Me), Arg(Me)2, etc.); modification of carboxyl groups such as amidation or esterification; modification of thiol groups such as farnesylation, geranylation, and palmitoylation; glycosylation such as N-linked glycosylation to the nitrogen atom of the side chain of asparagine and O-linked glycosylation to the hydroxyl group of serine or threonine; PEG (polyethylene glycol); lipidation; acylation; hydroxylation of proline or lysine; and the like. The modification may be, for example, a post-translational modification. When the vasodilator of the present invention contains the modified peptide, it is preferable that the modified peptide contains, for example, hydroxyproline as a constituent amino acid.

[0020] The vasodilator of the present invention may contain only a collagen-derived tripeptide, only a collagen-derived dipeptide, or both as an active ingredient. In addition, the vasodilator of the present invention may contain, for example, one type of peptide or multiple types of peptides.

[0021] The vasodilator of the present invention may include, as the tripeptide or dipeptide, an isolated product, purified product, or crudely purified product of the tripeptide or dipeptide, or may include, for example, a proteolytic enzyme-treated product of a collagen-derived protein containing the tripeptide or dipeptide.

[0022] The collagen-derived tripeptide or collagen-derived dipeptide in the present invention is, for example, a tripeptide or dipeptide derived from collagen, and may be prepared from raw materials such as natural products or processed products thereof, or may be synthesized by a conventionally known synthesis method. The synthesis method may be, for example, a solid-phase synthesis method or a liquid-phase synthesis method, and may include a peptide synthesis method utilizing the reverse reaction of hydrolases, a peptide synthesis method using an organic chemical method, and a synthesis method using a genetic engineering method.

[0023] Examples of the preparation method from the raw material include purification from the raw material (collagen) itself or from a decomposition product of the raw material. The decomposition method of the raw material is not particularly limited, and can be performed, for example, by using an acid, an alkali, or a protease. In addition, the purification method can be a conventionally known separation and purification method that utilizes, for example, differences in adsorption affinity to an adsorbent, differences in solubility in various solvents, differences in elution rate due to the molecular sieve effect, and the like. In addition, extraction and concentration can be appropriately combined. The origin of the collagen is not particularly limited, and for example, it may be derived from an animal or may be synthesized by a conventionally known synthesis method. The animal is not particularly limited, and for example, it may be a human or a non-human animal. The non-human animal is, for example, a mammal other than a human, a bird, a reptile, an amphibian, a fish, and the like. The animal is preferably, for example, a fish from the viewpoint of the risk of infection, the odor after formulation, and the like. In this case, the vasodilator of the present invention includes, for example, a processed product obtained by treating fish-derived collagen with a protease.

[0024] The vasodilator of the present invention is, for example, in vivo You can also use in vitro The vasodilator of the present invention may be composed of, for example, a plurality of components. In this case, the vasodilator of the present invention may be called, for example, a vasodilator composition.

[0025] There are no particular limitations on the subjects to which the vasodilator of the present invention is administered. in vivoIn the case of use in the above, the subject of administration can be, for example, a human or a non-human animal other than a human. Examples of the non-human animal include a mouse, a rat, a rabbit, a dog, a sheep, a horse, a cat, a goat, a monkey, a guinea pig, etc. in vitro When used in the above, the subject of administration can be, for example, a cell, a tissue, an organ, etc. Examples of the cells can be, for example, cells collected from a living body, cultured cells, etc., and examples of the tissues can be, for example, oral mucosa such as gingiva. When the vasodilator of the present invention is administered to the oral mucosa, it can dilate, for example, blood vessels in the gingiva.

[0026] The conditions for use (administration conditions) of the vasodilator of the present invention are not particularly limited, and the administration form, administration time, administration amount, etc. can be appropriately set depending on, for example, the type of administration subject, etc.

[0027] The dosage of the vasodilator of the present invention is not particularly limited. in vivoWhen used in the above, the dosage can be appropriately determined depending on, for example, the type, symptoms, age, and administration method of the subject to be administered. As a specific example, when the vasodilator of the present invention contains the collagen-derived tripeptide, the total dosage of the tripeptide per day when administered to humans is, for example, 40 to 400 mg, 40 to 375 mg, 40 to 350 mg, 40 to 300 mg, 40 to 250 mg, 40 to 200 mg, 40 to 80 mg, or 37.5 to 75 mg. The number of times of administration per day is, for example, 1 to 6 times, 1 to 4 times, 1 to 3 times, or 1 to 2 times, and preferably 1 to 3 times. When the vasodilator of the present invention contains the collagen-derived dipeptide, the total dose of the dipeptide per day when administered to humans is, for example, 40 to 400 mg, 40 to 375 mg, 40 to 350 mg, 40 to 300 mg, 40 to 250 mg, 40 to 200 mg, 40 to 80 mg, or 37.5 to 75 mg. The number of times of administration per day is, for example, 1 to 6 times, 1 to 4 times, 1 to 3 times, or 1 to 2 times, and preferably 1 to 3 times. When the vasodilator of the present invention contains the collagen-derived tripeptide and the collagen-derived dipeptide, the total dose of the tripeptide and the dipeptide per day when administered to humans is, for example, 40 to 400 mg, 40 to 375 mg, 40 to 350 mg, 40 to 300 mg, 40 to 250 mg, 40 to 200 mg, 40 to 80 mg, or 37.5 to 75 mg. The number of times of administration per day is, for example, 1 to 6 times, 1 to 4 times, 1 to 3 times, or 1 to 2 times, and preferably 1 to 3 times. The content of the collagen-derived tripeptide or collagen-derived dipeptide in the vasodilator of the present invention is not particularly limited and can be appropriately set according to, for example, the daily dosage.

[0028] The administration form of the vasodilator of the present invention is not particularly limited. in vivoWhen the vasodilator is administered intravenously, it may be administered orally or parenterally. Examples of the parenteral administration include intravenous injection (intravenous administration), intramuscular injection (intramuscular administration), transdermal administration, subcutaneous administration, intradermal administration, enteral administration, rectal administration, vaginal administration, nasal administration, pulmonary administration, intraperitoneal administration, and topical administration. Since the vasodilator of the present invention can dilate blood vessels by penetrating, for example, the oral mucosa, it is preferably administered by oral administration such as transmucosal administration, and more preferably by oral mucosal administration.

[0029] The dosage form of the vasodilator of the present invention is not particularly limited and can be appropriately determined depending on, for example, the administration form. Examples of the dosage form include liquid and solid forms. Specific examples of the dosage form include preparations for oral administration such as release-modified preparations (enteric preparations, sustained-release preparations, etc.), capsules, oral liquids (elixirs, suspensions, emulsions, aromatic perfumes, lemonades, etc.), syrups (syrup preparations, etc.), granules (effervescent granules, fine granules, etc.), powders, tablets (orally disintegrating tablets, chewable tablets, effervescent tablets, dispersible tablets, dissolving agents, coated tablets, etc.), pills, and oral jellies; preparations for oral administration such as oral tablets (gums, sublingual tablets, troches, drops, buccal tablets, adhesive tablets, etc.), oral sprays, oral semisolids, and mouthwashes; injections (implanted injections, sustained-release injections, infusions (infusion preparations, etc.), freeze-dried injections, powder injections, pre-filled syringes, preparations for bronchial and pulmonary application such as inhalants (inhalation aerosols, inhalation liquids, inhalation powders, etc.); preparations for ocular administration such as eye ointments and eye drops; preparations for ear administration such as ear drops; preparations for nasal application such as nasal drops (nasal liquids, nasal powders, etc.); preparations for rectal application such as suppositories, semi-solid rectal preparations, and enemas; preparations for vaginal application such as vaginal suppositories and vaginal tablets; preparations for skin application such as external liquid preparations (alcohol preparations, liniments, lotions, etc.), creams, gels, external solid preparations (external powders, etc.), sprays (external aerosols, pump sprays, etc.), patches (tapes, poultices, etc.), and ointments. When the vasodilator of the present invention is orally administered, the dosage form may be, for example, a tablet, a coated tablet, a pill, fine granules, granules, powder, capsule, liquid, syrup, emulsion, suspension, etc. When the vasodilator of the present invention is parenterally administered, the dosage form may be, for example, a preparation for injection, a preparation for drip infusion, etc. When the vasodilator of the present invention is percutaneously administered, the dosage form may be, for example, a topical agent such as a patch, a liniment, an ointment, a cream, or a lotion.

[0030] The vasodilator of the present invention may contain additives, if necessary. When the vasodilator of the present invention is used as a medicine or pharmaceutical composition, the additives preferably contain pharma- ceutical acceptable additives or pharma-ceutical acceptable carriers. The additives are not particularly limited, and examples thereof include base materials, excipients, colorants, lubricants, binders, disintegrants, stabilizers, preservatives, flavorings such as fragrances, etc. In the vasodilator of the present invention, the amount of the additives is not particularly limited as long as they do not interfere with the function of the collagen-derived tripeptide or collagen-derived dipeptide.

[0031] Examples of the excipient include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, and dextrin; cellulose derivatives such as crystalline cellulose; organic excipients such as gum arabic; dextran; and pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, and magnesium aluminometasilicate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; and sulfates such as calcium sulfate. Examples of the lubricant include metal stearates such as stearic acid, calcium stearate, and magnesium stearate; talc; polyethylene glycol; silica; and hydrogenated vegetable oil. Examples of the flavoring agent include flavors such as cocoa powder, peppermint, aromatic powder, peppermint oil, borneol, and cinnamon powder, sweeteners, and acidulants. Examples of the binder include hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone, macrogol, etc. Examples of the disintegrant include cellulose derivatives such as carboxymethyl cellulose and calcium carboxymethyl cellulose; chemically modified starches and chemically modified celluloses such as carboxymethyl starch, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone, etc. Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; sorbic acid, etc.

[0032] The collagen-derived tripeptide or collagen-derived dipeptide may be commercially available or may be prepared in-house as described above.

[0033] As described above, the vasodilator of the present invention can dilate blood vessels. Therefore, the vasodilator of the present invention can improve, for example, the blood flow of a subject, and can promote the reduction and / or alleviation of symptoms associated with blood circulation function. The vasodilator of the present invention can also be used as an additive in, for example, cosmetics, foods, etc.

[0034] The vasodilator of the present invention exhibits vasodilatory action, and therefore can improve, for example, blood flow. Therefore, the vasodilator of the present invention can be used as a medicine for diseases caused by vascular contraction or constriction. Examples of diseases caused by vascular contraction or constriction include periodontal disease, high blood pressure, cerebral hemorrhage, stiff shoulders, headache, cerebral circulation, blood congestion, arteriosclerosis, cerebral infarction, bedsores, itching due to dryness, wrinkles, skin disorders due to ultraviolet rays, atopic dermatitis, osteoporosis, tendon rupture, osteoarthritis of the knee, etc.

[0035] The vasodilator of the present invention may be contained in, for example, foods, supplements, etc. that are taken on a daily basis.

[0036] <Periodontal disease treatment drug> As described above, the periodontal disease treatment drug of the present invention contains the vasodilator of the present invention. The periodontal disease treatment drug of the present invention is characterized by containing the vasodilator of the present invention, i.e., at least one of a collagen-derived tripeptide or a collagen-derived dipeptide, and other configurations and conditions are not particularly limited. In the periodontal disease, for example, blood vessels are constricted due to a chronic inflammatory reaction of the gums, and the alveolar bone that supports the periodontal tissue is resorbed. Since the periodontal disease treatment drug of the present invention contains the vasodilator of the present invention, it can dilate the blood vessels of the gums by administering it to the oral mucosa, and as a result, periodontal disease can be treated. The periodontal disease treatment drug of the present invention can be described in the same manner as the vasodilator of the present invention.

[0037] In the present invention, examples of the periodontal disease include gingivitis, periodontitis, and alveolar pyorrhea.

[0038] In the present invention, "treatment" may mean any of suppression or prevention of disease onset, suppression or halt of disease progression, suppression or halt of disease symptom progression, and / or improvement of disease. Therefore, the periodontal disease therapeutic agent of the present invention can be referred to as, for example, a periodontal disease inhibitor, preventive agent, progression inhibitor, progression halt agent, and / or ameliorator. Furthermore, the periodontal disease therapeutic agent of the present invention may (significantly) suppress the symptoms or progression of disease compared to the absence of the periodontal disease therapeutic agent of the present invention (non-administration condition), and may progress the disease compared to the start (start of administration).

[0039] The administration conditions of the periodontal disease therapeutic agent of the present invention are not particularly limited, and for example, the administration form, administration time, administration amount, etc. can be appropriately set depending on the type of administration subject, etc. For the administration subject and administration conditions of the periodontal disease therapeutic agent of the present invention, for example, the explanation of the administration subject and administration conditions for the vasodilator of the present invention can be used.

[0040] <Oral composition> As described above, the oral composition of the present invention contains the vasodilator of the present invention. The oral composition of the present invention is characterized by containing the vasodilator of the present invention, i.e., at least one of a collagen-derived tripeptide or a collagen-derived dipeptide, and other configurations and conditions are not particularly limited. The oral composition of the present invention can dilate blood vessels because it contains the vasodilator of the present invention. The oral composition of the present invention can be described in the same manner as the vasodilator of the present invention.

[0041] The oral cavity composition of the present invention may contain, for example, other components. The other components are not particularly limited, and examples thereof include a gum base, a sweetener, a flavoring, an acidulant, a coloring agent, and the like.

[0042] <Food Composition> As described above, the food composition of the present invention contains the vasodilator of the present invention. The food composition of the present invention is characterized by containing the vasodilator of the present invention, i.e., at least one of a collagen-derived tripeptide or a collagen-derived dipeptide, and other configurations and conditions are not particularly limited. The food composition of the present invention can dilate blood vessels because it contains the vasodilator of the present invention. The food composition of the present invention can be described in the same manner as the vasodilator of the present invention.

[0043] The food composition of the present invention can be used, for example, by adding it to food. Therefore, the food composition of the present invention can also be called, for example, a food additive. In the present invention, the "food" may be liquid or solid, and can also be called, for example, a food or drink product or a food or drink product. The food to which the food composition of the present invention is added is not particularly limited.

[0044] The amount of the food composition added to the food is, for example, 1 to 100% by weight, 10 to 100% by weight, or 15 to 100% by weight.

[0045] <Vasodilatory methods> As described above, the vasodilator method of the present invention includes an administration step of administering a vasodilator to a subject, and the vasodilator is the vasodilator of the present invention. The vasodilator method of the present invention is characterized by administering the vasodilator of the present invention to a subject, that is, administering at least one of a collagen-derived tripeptide or a collagen-derived dipeptide, and other steps and conditions are not particularly limited. The vasodilator method of the present invention administers the vasodilator of the present invention to a subject, and therefore can dilate the blood vessels of the subject. The vasodilator method of the present invention can be described with reference to the description of the vasodilator of the present invention.

[0046] In the vasodilatory method of the present invention, the administration step includes, for example, in vitro or in vivo For the subjects and conditions for administration of the vasodilator of the present invention, the explanation of the subjects and conditions for administration of the vasodilator of the present invention can be cited, for example.

[0047] <Periodontal disease treatment method> As described above, the method for treating periodontal disease of the present invention includes an administration step of administering a therapeutic drug for periodontal disease to a subject, and the therapeutic drug for periodontal disease is the therapeutic drug for periodontal disease of the present invention. The method for treating periodontal disease of the present invention is characterized by administering the therapeutic drug for periodontal disease of the present invention to a subject, i.e., administering at least one of a collagen-derived tripeptide or a collagen-derived dipeptide, and other steps and conditions are not particularly limited. The method for treating periodontal disease of the present invention administers the therapeutic drug for periodontal disease of the present invention to a subject, and therefore can treat periodontal disease in the subject. The explanation of the therapeutic drug for periodontal disease and the vasodilator of the present invention can be used for the method for treating periodontal disease of the present invention.

[0048] In the periodontal disease treatment method of the present invention, the administration step includes, for example, in vitro or in vivo For the subjects and conditions for administration of the periodontal disease remedy of the present invention, the explanation for the subjects and conditions for administration of the vasodilator of the present invention can be cited, for example.

[0049] <Use of agent> The present invention relates to at least one of a collagen-derived tripeptide and a collagen-derived dipeptide, or use thereof, for vasodilatation, and at least one of a collagen-derived tripeptide and a collagen-derived dipeptide, or use thereof, for treating periodontal disease. The present invention relates to at least one of a collagen-derived tripeptide and a collagen-derived dipeptide, or use thereof, for producing a vasodilator or a therapeutic agent for periodontal disease. For example, the above-mentioned explanations of the vasodilator, the method for vasodilatation of a therapeutic agent for periodontal disease, and the method for treating periodontal disease of the present invention can be used for the present invention.

[0050] Next, examples of the present invention will be described. However, the present invention is not limited to the following examples. Commercially available reagents were used according to their protocols unless otherwise specified. EXAMPLES

[0051] [Example 1] The vasodilator of the present invention is in vivo They confirmed that it can dilate blood vessels.

[0052] (Preparation of peptide fractions) Collagen tripeptide (CTP, FACT-P, Gly-Pro-Hyp, manufactured by MC Plot Biotechnology, Inc.) was dissolved in physiological saline to give a CTP concentration of 0.5 g / ml, to prepare CTP-containing physiological saline.

[0053] Female rats (Wistar, n=1) aged 8 weeks or older were anesthetized by subcutaneous administration of urethane (Sigma-Aldrich) at 1.75 g / kg body weight. The rats were then fixed to an observation table. After the fixation, the observation table was placed under a microscope (Nikon SMZ25, Nikon). In order to prevent the gum surface from drying out, an ultrasonic warm air inhaler (humidifier, UN-133B, A&D) was operated in mist-free mode during the test to spray moist air (relative humidity: 95%). Figures 1(A) and (B) show photographs of the rat mandibular incisor gingiva. Figure 1(A) is a photograph of the entire observation site, and Figure 1(B) is an enlarged photograph of the blood vessel observation area surrounded by a white frame in Figure 1(A). The blood vessel observation area included veins of a thickness described below, and the part in focus in the image was selected.

[0054] As a control, 10 μl of saline was dropped onto the gingiva of the rat's lower incisor, and the subcutaneous tissue of the blood vessel observation area (area enclosed in a square in Figure 1 (A)) of the lower incisor gum was photographed and recorded under a microscope for 30 minutes after the drop. After that, after a 5-minute interval, the saline containing CTP was dropped onto the same site, and the subcutaneous tissue was photographed and recorded under a microscope for 30 minutes after the drop.

[0055] Images were cut out from the obtained movie, and the vascular diameter before the administration of physiological saline was measured using the scale tool of Photoshop (registered trademark, Adobe). Based on the vascular diameter before the administration of physiological saline, the size of the blood vessel was classified into three stages (thick: about 25 μm, medium: 15 to 20 μm, thin: about 10 μm) as shown in FIG. 1(B). Then, the vascular diameter of the blood vessel of each classification was measured every 5 minutes after the administration of physiological saline or CTP from the captured images of the movie. These results are shown in FIG. 2.

[0056] 2(A)-(C) are graphs showing the change in vascular diameter after administration of the vasodilator of the present invention or physiological saline, where (A) shows the results for medium-sized blood vessels (CTP1, control1), (B) shows the results for large blood vessels (CTP2, control2), and (C) shows the results for small blood vessels (CTP3, control3). In FIG. 2(A)-(C), the horizontal axis indicates the time elapsed after administration, and the vertical axis indicates the vascular diameter. As shown in FIG. 2(A)-(C), the addition of control physiological saline resulted in almost no change in vascular diameter compared to before addition. In contrast, when CTP was administered, the vascular diameter expanded at any time after administration compared to before CTP addition. In addition, when CTP was administered, the vascular diameter expanded regardless of the size of the blood vessel, but the degree of expansion of the vascular diameter was remarkable in smaller blood vessels (microvessels, CTP3). In addition, when CTP is administered in vivo, it is decomposed into proline-hydroxyproline. As shown in Figures 2(A) to (C), administration of CTP can dilate blood vessels, which indicates that proline-hydroxyproline can also dilate blood vessels.

[0057] From these results, the vasodilator of the present invention has the following properties: in vivo It has been found that it can dilate blood vessels.

[0058] [Example 2] The vasodilator of the present invention is in vivo It was confirmed that blood flow can be improved in

[0059] (1) Administration of CTP Eight healthy adults (four men and four women) were orally administered the vasodilator of the present invention (CTP-containing food (CTP 37.5 mg / 1 tablet): Kawaii Gri Collagen (registered trademark), Bioradical Institute) for a specified test period (14 days from October 28, 2020 to November 11, 2020), and changes in periodontal tissue blood flow before and 14 days after the start of administration were confirmed. Specifically, each subject was asked to take the vasodilator of the present invention twice a day (after breakfast and dinner), chewing five tablets per intake about 10 times per tablet, allowing the CTP in the vasodilator of the present invention time to contact the oral mucosa sufficiently, and then swallowing it. Note that on the start date of intake (administration) (October 28, 2020), intake was limited to after dinner, and on the end date of intake (administration) (November 11, 2020), intake was limited to after breakfast. In addition, each subject had the oral cavity cleaned with a specified oral care product (toothpaste: PC Clinica M (manufactured by Lion Corporation), toothbrush: DENT EX33 M (manufactured by Lion Corporation)) from 3 days before the start of the test period to the end of the test (October 25, 202 to November 11, 2020), and during the test period, 15 minutes after taking the vasodilator of the present invention, the subjects brushed their teeth with the oral care product. During the test period, the subjects were instructed to refrain from taking supplements other than the vasodilator of the present invention. If the subjects forgot to take the vasodilator of the present invention after breakfast or dinner, they did not take it additionally and recorded that they had forgotten to take it. In addition, if the subjects became unwell due to a cold or the like, or if they took any medicine, they were also asked to record that fact.

[0060] (2) Measurement of blood flow Before (October 28, 2020) and after (November 11, 2020) the vasodilator of the present invention was taken, the microvascular blood flow rate (Flow: ml / min / 100g) of the periodontal tissue of each subject was measured using a laser Doppler blood flow meter (Laser blood flow meter ALF21N, manufactured by Advance Co., Ltd.). Specifically, a vinyl tube tip with a distance from the tip of the tip to the tip of the probe of the blood flow meter (Laser blood flow meter probe [non-contact type] NCD type, manufactured by Advance Co., Ltd.) of about 1.5 mm was joined to the tip of the blood flow meter. Then, the tip of the probe tip was lightly touched to the blood flow measurement point of each subject without pressing it against the blood flow. The measurement was performed five times for each measurement point, and the average value of the five measurements was taken as the blood flow rate at the measurement point. The tip was replaced for each subject, and when the tip was detached from the probe, the probe was lightly wiped with an alcohol wattle. The measurement points of the blood flow will be described with reference to FIG. 3. The teeth to be measured were the second and sixth teeth in the upper right (upper right 2, upper right 6), the fourth tooth in the lower right (lower right 4), the fourth tooth in the upper left (upper left 4), and the second and sixth teeth in the lower left (lower left 2, lower left 6), as shown in FIG. 3(A). As shown in FIG. 3(B), the measurement points were the attached gingiva located about 2 mm from the gingival-buccal transition of each measurement target tooth in the attached gingiva direction, and the middle part of the crown width of the target tooth mesiodistally. In addition, the measurement points were also similarly set for the labial gingiva (upper right 1) in the center connecting the labial cervical part of the maxillary right central incisor and the lateral incisor. The data was collected and recorded using a data logger (insulated 10-channel handy logger midi Logger GL240, manufactured by Graphtec Corporation). These results are shown in Table 1 below.

[0061] [Table 1]

[0062] As shown in Table 1, the ingestion of the vasodilator of the present invention improved the blood flow rate of the periodontal tissue microvessels of each subject. When measured by site, the blood flow rate was statistically significantly improved in the upper right 1 and lower left 6. This shows that the vasodilator of the present invention can improve the blood flow of the subjects.

[0063] [Example 3] The vasodilator of the present invention is in vivo It was confirmed that this treatment is effective in treating periodontal disease.

[0064] In the same manner as in Example 2(1) above, the vasodilator of the present invention was orally administered to the subjects, and before and after the specified test period, periodontal pocket examinations (PD: Probing Depth (mm)), the presence or absence of gingival bleeding (BOP: Bleeding on Probing), and gingival index examinations (GI: Gingival Index) were performed.

[0065] (1) Periodontal pocket examination The marginal gingiva of the tooth to be measured was probed with a periodontal probe equipped with a scale to confirm the depth of the periodontal pocket (Probing Depth (mm)).

[0066] (2) Presence or absence of gingival bleeding The marginal gingiva of the tooth being measured was probed with a periodontal probe. If bleeding from the gingiva was observed within 30 seconds, it was evaluated as having gingival bleeding (Y: BOP+), and if no bleeding was observed, it was evaluated as not having gingival bleeding (N: BOP-).

[0067] (3) Gingivitis Index Test The marginal gingiva of the four surfaces (buccal / labial surface, lingual / palatal surface, mesial surface, and distal surface) shown in Figure 4 of the teeth being measured was scored according to the following evaluation criteria, and the average of the evaluations of the four surfaces was used as the gingival index (GI) of each tooth being measured. Gingivitis Index Rating 0: Macroscopic findings are normal, no bleeding on probing. 1: Mild redness and inflammation of the gingiva is visible to the naked eye, but there is no bleeding on probing. 2: Moderate redness, inflammation, and swelling of the gingiva are visible to the naked eye, and bleeding occurs when probing. 3: Severe redness, inflammation, and swelling of the gingiva are visible to the naked eye, and bleeding is observed not only on probing but also spontaneously.

[0068] The results of the periodontal pocket examination are shown in Table 2 below, the results of the presence or absence of gingival bleeding are shown in Table 3 below, and the results of the gingivitis index test are shown in Table 4 below. Note that for subject H, a problem occurred during the periodontal pocket examination, so the periodontal pocket examination was not performed on subject H.

[0069] [Table 2]

[0070] [Table 3]

[0071] [Table 4]

[0072] As shown in Tables 2 to 4, each subject showed improvement in periodontal pockets, gingival bleeding, and gingivitis index by taking the vasodilator of the present invention. Therefore, it was found that the vasodilator of the present invention can treat gingivitis and periodontal disease in subjects.

[0073] Although the present invention has been described above with reference to the embodiments and examples, the present invention is not limited to the above-mentioned embodiments and examples. Various modifications that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

[0074] This application claims priority based on Japanese Patent Application No. 2019-216078, filed on November 29, 2019, the disclosure of which is incorporated herein in its entirety.

[0075] <Additional Notes> Some or all of the above-described embodiments and examples may be described as follows, but are not limited to the following: (Appendix 1) A vasodilator comprising at least one of a collagen-derived tripeptide and a collagen-derived dipeptide. (Appendix 2) The collagen-derived tripeptide includes a tripeptide consisting of glycine and any two amino acids, The vasodilator described in Appendix 1, wherein the any amino acid is an amino acid selected from the group consisting of proline, hydroxyproline, glutamic acid, alanine, and arginine. (Appendix 3) 3. The vasodilator according to claim 1 or 2, wherein the collagen-derived tripeptide comprises a tripeptide consisting of glycine, proline, and hydroxyproline. (Appendix 4) The vasodilator according to any one of claims 1 to 3, wherein the collagen-derived dipeptide contains at least two amino acids selected from glycine, proline, hydroxyproline, glutamic acid, alanine, and arginine. (Appendix 5) The vasodilator according to any one of claims 1 to 4, wherein the collagen-derived dipeptide comprises a dipeptide composed of proline and hydroxyproline. (Appendix 6) 6. The vasodilator according to any one of claims 1 to 5, wherein at least one of the collagen-derived tripeptide and the collagen-derived dipeptide comprises a product obtained by treating fish-derived collagen with a protease. (Appendix 7) 7. The vasodilator according to any one of claims 1 to 6, wherein at least one of the amino acid residues constituting the collagen-derived tripeptide and the collagen-derived dipeptide is a natural amino acid residue or a modified amino acid residue. (Appendix 8) The vasodilator according to any one of claims 1 to 7, wherein the vasodilator is a transmucosal vasodilator. (Appendix 9) The vasodilator according to any one of claims 1 to 8, wherein the vasodilator is an oral mucosal vasodilator. (Appendix 10) A therapeutic agent for periodontal disease comprising a vasodilator according to any one of claims 1 to 9. (Appendix 11) An oral composition comprising a vasodilator according to any one of claims 1 to 9. (Appendix 12) A food composition comprising a vasodilator according to any one of claims 1 to 9. [Industrial Applicability]

[0076] As described above, the vasodilator of the present invention contains at least one of a collagen-derived tripeptide and a collagen-derived dipeptide, and therefore can dilate blood vessels. Therefore, the vasodilator of the present invention can improve, for example, the blood flow of a subject, and can promote the reduction and / or alleviation of symptoms associated with blood circulation function. Therefore, the vasodilator of the present invention is extremely useful, for example, in the medical field.

Claims

1. A vasodilator comprising at least one of a collagen-derived tripeptide and a collagen-derived dipeptide.

2. The collagen-derived tripeptide includes a tripeptide consisting of glycine and any two amino acids, The vasodilator of claim 1, wherein the optional amino acid is an amino acid selected from the group consisting of proline, hydroxyproline, glutamic acid, alanine, and arginine.

3. The vasodilator according to claim 1 or 2, wherein the collagen-derived tripeptide comprises a tripeptide consisting of glycine, proline, and hydroxyproline.

4. The vasodilator according to claim 1 , wherein the collagen-derived dipeptide contains at least two amino acids selected from glycine, proline, hydroxyproline, glutamic acid, alanine, and arginine.

5. The vasodilator according to claim 1 , wherein the collagen-derived dipeptide comprises a dipeptide composed of proline and hydroxyproline.

6. The vasodilator according to claim 1 , wherein at least one of the collagen-derived tripeptide and the collagen-derived dipeptide comprises a product obtained by treating fish-derived collagen with a protease.

7. The vasodilator according to claim 1 , wherein at least one of the amino acid residues constituting the collagen-derived tripeptide and the collagen-derived dipeptide is a natural amino acid residue or a modified amino acid residue.

8. The vasodilator according to claim 1 , wherein the vasodilator is a transmucosal vasodilator.

9. The vasodilator according to claim 1 , wherein the vasodilator is an oral mucosal vasodilator.

10. A therapeutic agent for periodontal disease comprising a vasodilator according to any one of claims 1 to 9.

11. An oral composition comprising a vasodilator according to any one of claims 1 to 9.

12. A food composition comprising a vasodilator according to any one of claims 1 to 9.

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