JP2007204368A - NF-kappaB ACTIVITY INHIBITOR COMPRISING SILK PEPTIDE AS ACTIVE INGREDIENT AND ORAL COMPOSITION - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a substance that is useful as a food material and has a high safety and NF-κB activity inhibitory action. <P>SOLUTION: The NF-κB activity inhibitor comprises a silk peptide, especially a silk peptide made into a liposome, as an active ingredient. The useful oral composition has preventing or ameliorating action on vascular lesion caused by metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipemia, neutral-fat lipemia, hypercholesterolemia, hypertension, obesity or arteriosclerosis. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  NF-κB activity inhibitor containing silk peptide, metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteriosclerosis It is related with the useful oral composition which has the prevention or improvement effect of the vascular disorder resulting from this.

  It is known that silk is composed of a protein mainly composed of fibroin and sericin, and a silk peptide that is a degradation product thereof is edible (see, for example, Patent Documents 1 and 2).

  Diabetes causes very characteristic vascular cell changes, increased endothelial cells (angiogenesis) and decreased pericytes (pericyte loss). It is known that this occurs in micro blood vessels such as retina, nerve, kidney, heart, periodontal, and small blood vessels. Glycated protein (AGE) is a general term for molecules generated by non-enzymatic glycation and modification of proteins, and it is known that AGE is generated and accumulated with age in human blood vessels. Recently, AGE promotes endothelial cell proliferation and delays pericyte proliferation, and it has been elucidated that it plays a major role as an aversion factor for diabetic angiopathy. Currently, two mechanisms are known for the mechanism of action by which AGE causes damage to vascular cells. One is that the extracellular matrix protein, membrane protein, or intracellular protein is glycosylated, resulting in the breakdown of the functions of these proteins and the cellular functions dependent on them. The other is a cellular response caused by a receptor having AGE as a ligand, and several AGE receptors such as RAGE (Receptor for AGE), MSR-A, and LOX-1 have been reported so far. Among them, the AGE-RAGE system has been proved to be deeply involved in the hate of diabetic angiopathy from studies using RAGE knockout mice.

  On the other hand, it has been reported that NF-κB is deeply involved in vascular disorders exhibited by AGE. It is believed that AGE enhances intracellular oxidative stress via RAGE, and that transcription factor NF-κB (Nuclear Factor-κB) is activated via the Ras / MAP kinase pathway in a oxidative stress-dependent manner. NF-κB was first identified in 1986 as a transcription factor that binds to the B fragment of the enhancer region required for the immunoglobulin K light chain gene to be expressed specifically in mature B cells. After that, inactive NF-κB forms a complex with a protein called I-κB and is localized in the cytoplasm, and I-κB is dissociated by activation stimulation from the outside of the cell. It was revealed that NF-κB translocates to the nucleus and induces gene expression. Therefore, NF-κB is a transcription factor as well as a signal transduction factor that connects the cytoplasm and nucleus. Activation of NF-κB causes the production of IL-1 and TNF-α, which are inflammatory cytokines. Furthermore, activation of vascular endothelium and vascular smooth muscle, proliferation of vascular endothelium and vascular smooth muscle, enhancement of inflammatory response In addition, expression of adhesion molecules such as VCAM-1 and ICAM-1 on vascular endothelial cells and production of chemotactic factors such as IL-8 and MCP-1 resulted in leukocytes (monocytes and neutrophils). ), Adhesion of leukocytes to vascular endothelial cells, migration between vascular endothelial cells, infiltration of inflammatory lesions, and vascular sclerotic lesions are formed. Therefore, substances that suppress the activation of NF-κB in vascular endothelial cells are metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or It is a key substance for the prevention, treatment and delay of progression of vascular disorders caused by arteriosclerosis.

Cilostazol, an antithrombotic agent, is known as a substance that inhibits the activity of NF-κB, but adverse effects have occurred as a side effect of combined use with hemolytic agents, particularly diabetes and impaired glucose tolerance. It is difficult for patients to use.
Japanese Patent Laid-Open No. 02-177864 JP-A-11-139986

  Therefore, the purpose is to prevent or improve vascular disorders caused by metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis, etc. Therefore, it has been expected to develop a substance that inhibits the activity of NF-κB and has little risk of side effects. Accordingly, an object of the present invention is to provide a highly safe substance that can be used as a food material and has a NF-κB activity inhibitory action.

  The inventor screened a substance that inhibits the activation of NF-κB, found that the silk peptide and its liposomal product strongly inhibit the NF-κB activity, and completed the present invention.

That is, the present invention relates to the following NF-κB activity inhibitor and oral composition.
Item 1. NF-κB activity inhibitor containing a silk peptide.
Item 2. Item 2. The NF-κB activity inhibitor according to Item 1, further comprising a phospholipid.
Item 3. Item 3. The NF-κB activity inhibitor according to Item 2, wherein the silk peptide is made into a liposome.
Item 4. An oral composition containing the NF-κB activity inhibitor according to any one of Items 1 to 3.
Item 5. Metabolic syndrome containing silk peptide as an active ingredient, prevention of vascular disorders caused by diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteriosclerosis Oral composition for improvement.
Item 6. Item 6. The oral composition according to Item 5, further comprising a phospholipid.
Item 7. Item 7. The oral composition according to Item 6, wherein the silk peptide is made into a liposome.
Item 8. Item 8. The oral composition according to any one of Items 4 to 7, further comprising at least one selected from the group consisting of resveratrols, collagen, collagen derivatives, and n-3 unsaturated fatty acids.
Item 9. Item 9. The oral composition according to any one of Items 4 to 8, which is a food.
Item 10. Item 9. The oral composition according to any one of Items 4 to 8, which is a pharmaceutical.
Item 11. Item 11. The oral composition according to any one of Items 4 to 10, wherein the silk peptide is ingested in an amount of 50 mg to 1500 mg / day.
Item 12. Use of the silk peptide for manufacturing the food oral composition of claim | item 9.

  In the present invention, the silk peptide suppresses the activation of NF-κB in vascular endothelial cells. By inhibiting the activation of NF-κB, it is possible to suppress vascular injury caused by enhancement of AGE intracellular oxidative stress via RAGE. Therefore, the present invention can prevent or prevent vascular disorders caused by metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis, etc. Useful for improvement.

  The NF-κB activity inhibitor and oral composition of the present invention contain a silk peptide as an active ingredient. The silk peptide can be used without particular limitation as long as it is edible. A commercially available silk peptide can also be used. Further, for example, those produced by the method described in JP-A-11-139986 can also be used. That is, silk fibroin protein or silk sericin protein prepared from silk is used as a raw material, and this raw material can be produced by acid, alkali and enzyme decomposition. Preferably, it is possible to use one obtained by decomposing heat-dissolved silk with a proteolytic enzyme and converting it into a peptide.

  The molecular weight of the preferred silk peptide is 300 to 15,000, more preferably 300 to 5,000. The silk peptide preferably has a high content of glycine and alanine in its amino acid composition. For example, the content of glycine and alanine is preferably 10 to 80% and more preferably 20 to 70%.

  The NF-κB activity inhibitor and oral composition of the present invention preferably further contain a phospholipid in addition to the essential silk peptide. By blending phospholipids, the effect of promoting absorption into the body can be expected. The blending amount of the phospholipid in the NF-κB activity inhibitor is usually about 1 to 80% by weight, preferably about 3 to 65% by weight, more preferably about 5 to 50% by weight. Moreover, the compounding quantity of the phospholipid in an oral composition is about 1 to 75 weight% normally, Preferably it is about 3 to 60 weight%, More preferably, it is about 5 to 45 weight%.

  Examples of phospholipids include phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, sphingomyelin, soy lecithin, sunflower lecithin, corn lecithin, cottonseed oil lecithin, egg yolk lecithin, egg white lecithin, rape seed lecithin and the like. Examples include lecithin, hydrogenated lecithin, phospholipid derivatives obtained by introducing polyethyleneglycol or aminoglycans into these phospholipids. These phospholipids can be used singly or in combination of two or more of these, soy lecithin, egg yolk lecithin, sunflower lecithin, partially hydrogenated soybean lecithin, partially hydrogenated egg yolk lecithin, partially Hydrogenated sunflower lecithin is preferred.

  In the present invention, the phospholipid is preferably formulated in the form of liposomes. For example, liposomal silk peptides are preferred. As the liposomal silk peptide, it is preferable that the silk peptide is enclosed in a space surrounded by the liposome membrane, but the silk peptide may be included as a liposome membrane constituent and constitutes a multilamellar liposome. It may be contained between multiple membranes, or may be contained in a form in which the silk peptide is attached or bound to the outermost membrane of the liposome membrane.

  Liposomes are lipid vesicles having a bilayer formed by hydrating lipids mainly composed of phospholipids with a sufficient amount of water. Liposomes can incorporate water-soluble drugs into the inner water layer and lipid-soluble drugs into the lipid bilayer, and they can be applied as drug carriers for DDS preparations for the purpose of drug targeting, sustained release, and reduction of side effects. Has been tried. Liposomes are also known to have high safety because they are composed of components of biological membranes.

  In general, liposomes are classified based on the number of lipid bilayers, and are classified into multilamellar liposomes (MLV) and unilamellar liposomes. Single-membrane liposomes are further classified into SUV (small unilamella vesicle), LUV (large unilamella vesicle), GUV (giant unilamella vesicle), etc. according to the size. Any of these can be used in the present invention. Preferred is MLV. In the present invention, the size of the liposome is usually 30 to 100 nm, preferably 30 to 600 nm, more preferably 50 to 400 nm.

  The liposomal silk peptide can be produced by a conventional method. For example, a desired amount of lecithin and, if necessary, a desired amount of sterol are solubilized with an appropriate organic solvent such as ethanol, and the solvent is removed under reduced pressure to form a membrane lipid. A liposome encapsulating a silk peptide can be obtained by adding an aqueous solution containing the physiologically active substance and stirring the mixture at about 1000 to 3000 rpm for about 2 to 5 minutes to prepare a liposome suspension.

  In addition to this method, a desired amount of lecithin and, if necessary, a desired amount of sterol are dissolved in a small amount of ethanol, dispersed in an aqueous solution or buffer solution, preliminarily emulsified, and then dispersed at high pressure. A liposome encapsulating a silk peptide can also be obtained by forming a multilayer and preparing a liposome suspension.

  The obtained suspension may be subjected to an operation for removing the silk peptide in the liposome outer liquid, for example, an operation for dialysis of the obtained filtrate after filtration of the suspension.

  The liposome suspension can be used in a liquid state, but can also be used as a lyophilized dry product. Liposomes can be in a form suitable for various oral intakes, including those obtained by tableting or encapsulating the dried product.

  The content of the silk peptide in the silked silk peptide is usually about 10 to 99% by weight, preferably about 20 to 95% by weight, more preferably about 30 to 90% by weight.

  As the lecithin used in the liposome, the phospholipids exemplified above can be used. Lecithin is also called phosphatidylcholine or 1,2-diacylglycerol 3-phosphocholine, and generally fatty acids are bound to the 1-position and 2-position of glycerol. In the present invention, it is preferable to use lecithin in which an unsaturated fatty acid having 12 to 24 carbon atoms is bonded to both or one of the 1st and 2nd positions in addition to the above exemplified lecithin. It is particularly preferable to use lecithin having a saturated fatty acid having ˜24 and an unsaturated fatty acid having 12 to 24 carbon atoms bonded to the 2-position. Here, the saturated fatty acid and the unsaturated fatty acid may be either linear or branched. As a preferable unsaturated fatty acid, an unsaturated fatty acid having 16 to 18 carbon atoms can be used. In particular, lecithin having a large amount of oleic acid and linoleic acid bonded to the 2-position is preferred. Specifically, egg yolk lecithin, soybean lecithin, and sunflower lecithin are preferable.

  The liposome of the present invention preferably contains a sterol in addition to lecithin. Sterols derived from animals such as cholesterol, lanosterol, dihydrolanosterol, desmosterol, dihydrocholesterol; plants such as β-sitosterol, campesterol, stigmasterol, brassicasterol, ergosterol, ergostadienol, sitosterol, brassicasterol Sterols derived from (phytosterol); sterols derived from microorganisms such as timosterol and ergosterol, and the like, can be used alone or in combination of two or more. Among these, cholesterol or phytosterol is preferably used.

  The molar ratio of lecithin and sterol in the liposome is preferably about 55:45 to 95: 5, more preferably about 60:40 to 90:10, and most preferably about 75:25 to 85:15. When the molar ratio is within these ranges, the stability of the liposome membrane is improved.

  The content of lecithin in the liposomal silk peptide is usually about 1 to 80% by weight, preferably about 3 to 65% by weight, and more preferably about 5 to 50% by weight.

  The sterol content in the liposomal silk peptide is usually about 0 to 40% by weight, preferably about 0.1 to 30% by weight, more preferably about 1 to 20% by weight.

  The content of lecithin or sterol can be measured by a known method. For example, the lecithin content can be quantified by Fiske-Subbarow method, and the sterol content can be quantified by HPLC, colorimetric method or the like.

  Furthermore, the surface of the silk peptide formed into a liposome can be coated, and this coating can also be used as an active ingredient. A preferable coating includes a coating with a polysaccharide containing a sulfate group. Examples of sulfate group-containing polysaccharides include fucoidan, carrageenan, agar, and heparin. The sulfate group-containing polysaccharide includes a sulfated polysaccharide that does not contain a sulfate group, and may be chondroitin sulfate, dermatan sulfate, or the like.

  As the sulfate group-containing polysaccharide, those having a molecular weight of about 5000 to 300,000 are preferably used. Among these sulfate group-containing polysaccharides, fucoidan and carrageenan can be preferably used, and fucoidan is particularly preferable.

  The amount of sulfate group-containing polysaccharide used is, for example, preferably about 10 to 500 parts by weight and more preferably about 20 to 200 parts by weight with respect to 100 parts by weight of lecithin contained in the liposome.

  The coating can be performed, for example, by adding a sulfate group-containing polysaccharide to a suspension containing a liposomal silk peptide and stirring at about 1000 to 3000 rpm for about 2 to 5 minutes. A plurality of liposomes may be contained in one coating film.

  The fact that the liposome is coated with the sulfate group-containing polysaccharide can be confirmed, for example, by changing the zeta potential of the liposome solution by adding and stirring the sulfate group-containing polysaccharide.

  In addition to lecithin and sterol, liposomal silk peptides include antioxidants such as tocopherol and ascorbic acid, organic acids such as lactic acid and citric acid, lipids such as phosphatidylglycerol and phosphatidylethanolamine, chitosan, Natural polymers such as fucoidan and hyaluronic acid, synthetic polymers such as polyethylene glycol and carboxyvinyl polymer, carbohydrates such as trehalose, lactulose and maltitol, polyols such as glycerin, and the like can be added.

  Moreover, in addition to the silk peptide, various substances can be encapsulated in the liposomal silk peptide as necessary. For example, sulfonylurea drugs such as tolbutamide, acetohexamide, tolazamide, glyclopyramide, glybazole, glibenclamide, gliclazide and glimepiride; biguanide drugs such as metformin hydrochloride and buformin hydrochloride; α-glucosidase inhibitors such as acarbose and voglibose; pioglitazone Thiazolidine derivatives; phenylalanine derivatives such as nateglinide; insulin and insulin derivatives (novolin R, humanin R, monotade, etc.); statin drugs such as pravastatin sodium and simvastatin (HMG-CoA reductase inhibitors); clofibrate, bezafibrate , Fibrates such as clofibrate aluminum, syncybrate, clinofibrate, losartan, candesartan, valsarta Angiotensin II receptor antagonists (AT1 receptor blocker: ARB) such as telmisartan and olmesartan; angiotensin converting enzyme inhibitors such as enalapril maleate, cetapril, captolyl, imidapril hydrochloride, cilazapril, delapril hydrochloride, lisinopril, quinapril hydrochloride (ACE inhibitor) Drugs).

  The content of the silk peptide in the NF-κB activity inhibitor of the present invention is usually about 10 to 99% by weight, preferably about 20 to 95% by weight, more preferably about 30 to 90% by weight. In addition, what is necessary is just to make it the amount of the silk peptide contained in a liposome become said range, when using the silk peptide made into the liposome.

  The content of the silk peptide in the oral composition of the present invention is appropriately set according to the form of the composition, etc., but is usually about 0.5 to 95% by weight, preferably about 0.8 to 80% by weight, more preferably About 1 to 80% by weight.

  In addition to the silk peptide, the oral composition of the present invention preferably contains at least one selected from the group consisting of resveratrols, collagen, collagen derivatives, and n-3 unsaturated fatty acids. These preferable compounding amounts in the oral composition are 0.8 to 80% by weight, and more preferable compounding amounts are 1 to 80% by weight. By blending these, an inflammation suppressing effect, a vascular disorder suppressing effect and the like can be expected.

  Examples of resveratrols are resveratrol, laponticine, polydatin, 3,4 ', 5-stilbenetriol-4'-glucoside, 2,3,5,4'-stilbenetetraol-2-glucoside, stilbamidine, 4-methoxystilbene, ε-viniferin, α-viniferin and the like, preferably resveratrol, polydatin, ε-viniferin, α-viniferin and the like. These resveratrols are contained in extracts such as aceae plants, vines, bay diatoms, mulberry, peanuts, and the like when resveratrols are added to the composition of the present invention. It is also possible to mix various extracts.

  Collagen is present in a great number of organisms from lower organisms to humans, and the collagen used in the present invention is not limited to its origin organism. For example, mammals such as cattle, pigs, sheep and goats, birds such as chickens and turkeys, fish such as tuna, bonito, crucian carp, eel, shark and ray, cartilaginous animals such as squid and octopus, arthropods Collagen obtained from tissues such as can be used. Collagen produced using a genetically modified animal can also be used.

  The type of collagen may be any type, but type I collagen, type II collagen, and type III collagen are preferred.

  Examples of the collagen derivative include substances obtained by treatments such as denaturation, hydrolysis, and modification of these collagens. Among them, a hydrolyzate of collagen is preferable, and its average molecular weight is preferably about 500 to 10,000, about 1,000 to 5,000, more preferably about 1,300 to 4,000, Most preferably, it is about 1,000.

  Examples of the n-3 unsaturated fatty acid include α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, octadecatetraenoic acid, eicosatetraenoic acid, docosapentaenoic acid, and preferably α-linolenic acid, Eicosapentaenoic acid, docosahexaenoic acid, and the like. n-3 unsaturated fatty acids can be used in perilla oil, linseed oil, fish oil, etc. When blending n-3 unsaturated fatty acids in the composition of the present invention, these oils are blended. You can also

  The oral composition of the present invention is preferably for pharmaceutical use or food use.

  The oral food composition of the present invention can be prepared by an ordinary method by combining silk peptide with an edible carrier, food material, food additive and the like according to the form of the food as required. As a form of the food, it can be used as a liquid food such as a beverage, a solid food such as a tablet, granule or chewable tablet. It can also be used as a semi-solid food such as yogurt. Specific food forms include juices, soft drinks, liquid drinks such as tea; powdered drinks such as powdered juice and powdered soup; chocolate, candy, chewing gum, ice cream, jelly, cookies, biscuits, cornflakes, chewable tablets Confectionery such as film sheets, wafers, gummi, rice crackers, buns; seasonings such as dressings and sauces; breads, noodles, konjac, kneaded products (kamaboko, etc.), sprinkles, oral spray, troches and the like.

  Examples of carriers for oral food compositions include sugar alcohols such as maltitol, xylitol, sorbitol, erythritol, excipients such as crystalline cellulose, lactose, sucrose, glucose, starch, carbonates, phosphates, gelatin, alginic acid, Antibacterial agents such as xanthan gum, cellulose, hydroxypropylcellulose, methylcellulose, carrageenan, pullulan, pectin, etc., sucrose fatty acid ester, sorbitan fatty acid ester, enzymatically treated lecithin, enzymatically decomposed lecithin, saponin, ascorbic acid, tocopherol, etc. Oxidizing agents, acidulants such as lactic acid, citric acid, gluconic acid and glutamic acid, vitamins, amino acids, fortifiers such as lactate, citrate and gluconate, fluidizing agents such as silicon dioxide, sucrose fatty acid esters , Stearates Lubricants, lactulose, raffinose, fructooligosaccharides, isomaltoligosaccharides, xylooligosaccharides, oligosaccharides such as soybean oligosaccharides, dietary fibers such as indigestible dextrin, psyllium, beet fiber, sucralose, acesulfame potassium, aspartame, glycyrrhizin, etc. Sweetener, mint oil, eucalyptus oil, cinnamon oil, fennel oil, clove oil, orange oil, lemon oil, rose oil, fruit flavor, mint flavor, peppermint powder, dl-menthol, l-menthol, etc. be able to. Further, as an additive, live bacteria such as lactic acid bacteria or dead bacteria, other probiotic materials, prebiotic materials, vitamins, herbal medicines, herbs and other plants themselves or extracts may be blended.

  The oral food composition of the present invention prevents metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis, etc. Or it can use for uses, such as a health food for improvement, a functional food, a food for specific health, a functional nutrition food, a food for a sick person. The food composition of the present invention comprises a silk peptide as an active ingredient, metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteries An indication that it is used to prevent or improve vascular disorders caused by sclerosis, or those with high blood sugar levels, those with high cholesterol, those with high neutral fat, those with high blood pressure, those with obesity It is possible to make a food with a label indicating that it is used by a person who has a thick waist or who is concerned about complications of diabetes.

  The oral composition of the present invention contains a silk peptide and, if necessary, a pharmaceutically acceptable carrier, and is a solid agent such as a liquid, tablet, granule, fine granule, powder, or the liquid or solid agent. Can be used as orally administrable pharmaceutical compositions such as capsules. Examples of pharmaceutically acceptable carriers include excipients and diluents. The oral pharmaceutical composition can also contain various additives such as fragrances. Such oral pharmaceutical compositions can prevent or prevent vascular disorders caused by metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteriosclerosis. It can be used as an improving agent.

  Examples of carriers include maltitol, lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, methylcellulose, glycerin, sodium alginate, gum arabic, talc, calcium monohydrogen phosphate, calcium hydrogen phosphate , Sodium hydrogen phosphate, dipotassium phosphate, potassium dihydrogen phosphate, calcium dihydrogen phosphate, sodium dihydrogen phosphate, calcium sulfate, calcium lactate, cacao butter, etc., simple syrup, glucose solution, starch Liquid, gelatin solution, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, cross polyvinyl pyrrolidone, hydroxypropyl cellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxy Tillcellulose, carboxyvinyl polymer, crystalline cellulose, powdered cellulose, crystalline cellulose / carmellose sodium, carboxymethylcellulose, shellac, methylcellulose, ethylcellulose, potassium phosphate, gum arabic powder, pullulan, dextrin, corn starch, pregelatinized starch, hydroxypropyl Binders such as starch, gelatin, xanthan gum, tragacanth, tragacanth powder, macrogol, dry starch, sodium alginate, agar powder, laminaran powder, sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearin Disintegrants such as acid monoglycerides, starches, lactose, etc., sucrose, stearic acid, cocoa butter, hydrogenated oils, etc. Inhibitors, absorption accelerators such as quaternary ammonium salts, sodium lauryl sulfate, humectants such as glycerin and starch, adsorbents such as starch, lactose, kaolin, bentonite, colloidal silicic acid, purified talc, stearate, Lubricants such as boric acid powder and polyethylene glycol can be used. Further, the tablets can be made into tablets with ordinary coatings as necessary, for example, sugar-coated tablets, gelatin-encapsulated tablets, enteric-coated tablets, film-coated tablets, double tablets, multilayer tablets and the like. Capsules are prepared by mixing the active ingredient with the various carriers exemplified above and filling them into hard gelatin capsules, soft capsules and the like.

  The liquid preparation may be an aqueous or oily suspension, solution, syrup, or elixir, and is prepared according to a conventional method using usual additives.

  Examples of flavoring agents or fragrances include mint oil, eucalyptus oil, cinnamon oil, fennel oil, clove oil, orange oil, lemon oil, rose oil, fruit flavor, mint flavor, peppermint powder, dl-menthol, and l-menthol. Etc.

  The intake amount of the NF-κB activity inhibitor and oral composition of the present invention is such that the intake amount of the silk peptide is usually about 50 to 1500 mg, preferably about 100 to 1000 mg per adult day. In addition, preferred administration subjects of the NF-κB activity inhibitor and oral composition of the present invention include those with impaired glucose tolerance (IGT), those with a medical condition induced by impaired glucose tolerance, and the like. Such administration subjects have vascular disorders due to metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis, etc. Or likely to have in the future. Glucose intolerance is a classification that is incorporated in the WHO Diabetes Diagnosis Criteria, and refers to a group with a fasting blood glucose level of less than 126 mg / dl and a 75 g glucose tolerance test (OGTT) 2-hour value of 140-199 mg / dl. It refers to symptoms that do not belong to either the diabetic or normal types.

  The NF-κB activity inhibitor and oral composition of the present invention can be used in the field of utilizing the NF-κB activity inhibitory action of an active ingredient silk peptide, such as metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, high It is useful for the prevention or improvement of vascular disorders caused by lipemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity, arteriosclerosis and the like. Vascular disorders occur in blood vessels throughout the body due to arteriosclerosis, vascular structural abnormalities, and deformities. The present invention includes these vascular disorders as targets of prevention or improvement, and among the vascular disorders, particularly arteriosclerosis such as brain, coronary artery, lower limb artery, and microvascular disorders in nerve tissue, retina, kidney tissue, periodontal tissue, etc. It is valid. Vascular disorders resulting from diabetes include diabetic complications such as retinopathy, nephropathy, neurosis, cataract, coronary artery disease, cerebral infarction, myocardial infarction, angina, foot gangrene, gingivitis, periodontal disease, etc. .

  Metabolic syndrome is closely related to the development of lifestyle-related diseases such as obesity, hypertension, cardiovascular disease, and obesity, high blood sugar, high triglycerides due to lack of exercise and high-calorie high-fat diet. It is said that risk factors such as diabetes, hypercholesterolemia, and hypertension overlap, and the combination of these risk factors increases the risk of developing diabetes, myocardial infarction, stroke, and the like. In April 2005, diagnostic criteria for metabolic syndrome were published. According to this diagnostic criteria, as a marker of visceral fat accumulation (visceral fat area of 100 square centimeters or more), which is an essential item, the waist circumference is 85 cm or more for men and 90 cm or more for women. Diagnosis of metabolic syndrome when two or more of the three items (1) to (3) are met. (1) Serum lipid abnormalities (triglyceride level 150 mg / dl or higher or HDL cholesterol level less than 40 mg / dl), (2) high blood pressure (highest blood pressure 130 mmHg or higher or minimum blood pressure 85 mmHg or higher), (3) hyperglycemia (fasting blood glucose level) 110mg / dl or more).

  The NF-κB activity inhibitor and oral composition of the present invention have an action of inhibiting the activity of NF-κB, and include metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, It is useful for preventing or improving vascular disorders caused by hypercholesterolemia, hypertension, obesity, arteriosclerosis and the like.

  Hereinafter, the present invention will be described in more detail with reference to examples and test examples, but the present invention is not limited to these examples.

Production Example 1: Preparation of Liposome Preparation of Silk Peptide Encapsulated Liposomes A liposome preparation encapsulating silk peptides was prepared by a thin film hydration method. Egg yolk lecithin and phytosterol were weighed so that the molar ratio was 7: 3 (egg yolk lecithin 63.0 mg, phytosterol 14.55 mg), and 6 ml of ethanol was added to dissolve this. The solvent was distilled off from this solution with a rotary evaporator to form a thin film. To this thin film, 6 ml of citrate buffer solution (pH 6.69) in which 180 mg of silk peptide was dissolved was added and stirred at 2000 rpm for about 3 minutes by a vortex mixer to obtain a liposome suspension. This suspension was filtered using a polycarbonate membrane having a pore size of 0.2 μm to obtain silk peptide-encapsulated liposomes. When this liposome was negatively stained by treatment with 1% phosphotungstic acid, dried and observed with an electron microscope, it was confirmed that multilamellar liposomes were formed.

Example 1: Inhibition of NF-κB activity in vascular endothelial cells The effect of silk peptide (Edible Silk, manufactured by Ichimaru Falcos, molecular weight of about 1000) and cilostazol (positive control) on NF-κB activity of vascular endothelial cells was tested. did. TNF-α is an NF-κB inducer, and when this is administered to cells, the cells activate NF-κB.
Normal human umbilical vein endothelial cells (HUVECs) passaged at passages 3-5 in FGF (fibroblast growth factor) -containing MCDB131 medium are suspended in a 60 mm collagen-coated dish at a concentration of 2 × 10 ⁵ cells / dish. 6 mL of the solution was implanted and cultured for 2 days at 37 ° C. and 5% CO ₂ (pre-culture).

  The medium was removed, and 6 mL of FGF-free MCDB131 medium containing the following test substance at a concentration of 20 μg / mL was added. However, the control was only the medium, cilostazol was dissolved in DMSO at a concentration of 30 mg / ml, and 4 μl was added to 6 ml of the medium to prepare a medium containing cilostazol at a concentration of 20 μg / ml.

  After culturing for 2 hours, in order to divide into TNF-α administration group and non-administration group, 3 μL of 10 μg / mL TNF-α was administered to the administration group so that the TNF-α concentration was 5 ng / mL. After further culturing for 2 hours, the medium was removed, 5 mL of cold PBS (−) was added, and the cells were collected in a Spitz tube with a cell scraper. After centrifugation (1500 rpm, 10 minutes, 4 ° C.) of the Spitz tube, the supernatant is removed, and 80 μL of the following cell lysate (TNF-α non-administered group) or 240 μL (TNF-α administered group) is added and suspended. Then, it was transferred to a 1.5 mL Eppendorf tube and left on ice for 10 minutes. Thereafter, centrifugation (12000 rpm, 20 minutes, 4 ° C.) was performed, and the supernatant was transferred to a new 1.5 mL Eppendorf tube and stored at −80 ° C. until measurement of NF-κB activity. NF-κB activity was measured using a Trans-AM NF-κB kit (NF-κB p50, Active Motif), which is a DNA-binding assay. The measurement results are shown in FIG.

<Medium>
FGF-containing MCDB131 medium :
A medium in which FGF, FBS, glutamine and antibiotics are added to MCDB131 medium (manufactured by GIBCO) at concentrations of 2 ng / mL, 10%, 10 mM and 1%, respectively.
FGF-free MCDB131 medium :
A medium with the same composition as the FGF-containing MCDB131 medium except that it does not contain FGF.
<Test substance>
None (control)
Silk peptide cilostazol <cell lysate>
The cell lysate was prepared by mixing the following reagents in the Trans-AM NF-κB kit.
Lysis Buffer [20 mM Hepes (pH 7.5), 350 mM NaCl, 20% glycerol, 1% lgepal-CA630, 1 mM MgCl ₂ , 0.5 mM EDTA, 0.1 mM EGTA]; 2955 μL
DTT (1M Dithiothreitol); 15μL
Protease inhibitor cocktail (Proprietary mix); 30μL

  Positive control cilostazol (p = 0.002) and silk peptide (p = 0.0008) suppressed the NF-κB activity of HUVECs by TNF-α stimulation. NF-κB activity was 87% with cilostazol (inhibition rate 13%) and 68% with silk peptide (inhibition rate 32%) when the control was 100%.

Example 2
The following substances were used as test substances and control solutions, and the cells were prepared in the same manner as in Example 1 except that a medium (4 μl → 6 ml) adjusted to a test substance concentration of 20 μg / ml was added. Culture, recovery and measurement of NF-κB activity were performed. The measurement results are shown in FIG.
<Test substance and control solution>
Citrate buffer (pH 6.7)
Silk peptide-containing citrate buffer (pH 6.7, containing silk peptide 30 mg / ml)
Silk peptide-encapsulated liposome-containing citrate buffer (pH 6.7, containing 30 mg / ml silk peptide)
DMSO
Cilostazol DMSO solution (30mg / ml)

  In the positive controls cilostazol (p = 0.1), silk peptide (p = 0.008), and silk peptide-encapsulated liposome (p = 0.002), NF-κB activity of HUVECs by TNF-α stimulation was suppressed. NF-κB activity is 94% for cilostazol (inhibition rate 6%), 95% for silk peptide (inhibition rate 5%), 60% for silk peptide-encapsulated liposomes when the control (DMSO or buffer) is 100%. The inhibition rate was 40%).

Production Example 2: Tablets Tablets were produced by a conventional method based on the following formulation.
Egg yolk lecithin 5.0% by weight
Vegetable sterol 0.8% by weight
Silk peptide 15.0% by weight
Crystalline cellulose 76.0% by weight
Sucrose fatty acid ester 3.2% by weight
100% by weight

Production Example 3: Tablets Tablets were produced by a conventional method based on the following formulation.
Silk peptide 30.0% by weight
Xylitol 22.0% by weight
Maltitol 36.0% by weight
Fructooligosaccharide 10.0% by weight
Magnesium stearate 1.0% by weight
Fragrance 1.0% by weight
100% by weight

Production Example 4: Granule A granule was produced by a conventional method based on the following formulation.
Soy lecithin 4.0% by weight
Vegetable sterol 1.0% by weight
Silk peptide 20.0% by weight
Lactose 30.0% by weight
Starch 43.0% by weight
Xanthan gum 1.0% by weight
Ascorbic acid 0.3% by weight
Fragrance 0.7% by weight
100% by weight

Production Example 5: Beverage A beverage was produced by a conventional method based on the following formulation.
Silk peptide 1.0% by weight
Citric acid 0.5% by weight
Sodium citrate 0.05% by weight
Fructose and glucose liquid sugar 10.0% by weight
Vitamin C 0.5% by weight
Fragrance 0.5% by weight
Purified water balance
100% by weight

Production Example 6: Capsule A capsule was produced by a conventional method based on the following formulation.
Sunflower lecithin 4.0% by weight
Cholesterol 0.8%
Silk peptide 17.0% by weight
DHA (docosahexaenoic acid) 1.0% by weight
Microcrystalline cellulose 49.0% by weight
Sucrose fatty acid ester 3.2% by weight
Hard capsule 25.0% by weight
100% by weight

Production Example 7: Beverage A beverage was produced by a conventional method based on the following formulation.
Sunflower lecithin 0.2% by weight
Silk peptide 1.0% by weight
Grape leaf extract 0.5% by weight
(Contains resveratrol)
Apple transparent concentrated fruit juice 10.0% by weight
Fructose and glucose liquid sugar 5.0% by weight
Citric acid crystals 0.2% by weight
Fragrance 0.2% by weight
L-ascorbic acid 0.1% by weight
Purified water balance
100% by weight

  The NF-κB activity inhibitor and oral composition of the present invention can be used in the field of utilizing the NF-κB activity inhibitory action of an active ingredient silk peptide, such as metabolic syndrome, diabetes, hyperglycemia, impaired glucose tolerance, high It can be used for the prevention or improvement of vascular disorders caused by lipemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteriosclerosis.

2 is a graph showing NF-κB activity of various test substances measured in Example 1. FIG. The vertical axis represents NF-κB (p50) activity, the horizontal axis represents the test substance name, (−) represents the TNF-α non-administered group, and (+) the TNF-α administered group. 2 is a graph showing NF-κB activity of various test substances measured in Example 2. FIG. The vertical axis represents NF-κB (p50) activity, the horizontal axis represents the test substance name, (−) represents the TNF-α non-administered group, and (+) the TNF-α administered group.

Claims (12)

NF-κB activity inhibitor containing a silk peptide. The NF-κB activity inhibitor according to claim 1, further comprising a phospholipid. The NF-κB activity inhibitor according to claim 2, wherein the silk peptide is made into a liposome. An oral composition containing the NF-κB activity inhibitor according to any one of claims 1 to 3. Metabolic syndrome containing silk peptide as an active ingredient, prevention of vascular disorders caused by diabetes, hyperglycemia, impaired glucose tolerance, hyperlipidemia, hypertriglyceremia, hypercholesterolemia, hypertension, obesity or arteriosclerosis Oral composition for improvement. Furthermore, the oral composition of Claim 5 containing a phospholipid. The oral composition according to claim 6, wherein the silk peptide is a liposome. The oral composition according to any one of claims 4 to 7, further comprising at least one selected from the group consisting of resveratrols, collagen, collagen derivatives, and n-3 unsaturated fatty acids. It is a foodstuff, The oral composition in any one of Claims 4-8. The oral composition according to any one of claims 4 to 8, which is a medicine. The oral composition according to any one of claims 4 to 10, wherein the silk peptide is ingested in an amount of 50 mg to 1500 mg / day. Use of a silk peptide for producing the oral food composition of claim 9.

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