JP5972235B2 - Blood triglyceride lowering agent - Google Patents

Description

  The present invention relates to a blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent, and a lipid metabolism improving agent.

  In recent years, people with dyslipidemia have reached 22 million people in Japan, including potential patients, due to bad lifestyle habits such as overeating, high fat diet, lack of exercise and so on. In addition, a latent patient means the human of the boundary of dyslipidemia and an appropriate value. Dyslipidemia refers to a disease in which there are too many lipids in the blood, specifically cholesterol and neutral fat. Hypercholesterolemia (high LDL-C blood) Symptom), and a condition with a high blood triglyceride concentration is called hypertriglyceridemia.

Triglyceride is a kind of neutral fat, and most of the neutral lipid contained in blood is triglyceride. It is known that dyslipidemia (so-called hyperlipidemia) is caused when a state of high triglyceride concentration continues in blood. Dyslipidemia is considered to be the cause of arteriosclerosis and is the first trigger to cause diseases such as heart disease and cerebrovascular disorder.
Currently, fibrates (for example, ethyl 2-methyl-2- (4-chlorophenoxy) propionate) are often used for the treatment of hypertriglyceridemia, but transient liver dysfunction is a side effect. Easy to cause.

Under such circumstances, it is desired to suppress an increase in blood triglyceride concentration by a method that is highly safe and has few side effects even when administered or ingested daily.
For example, Patent Document 1 proposes a blood triglyceride concentration increase inhibitor containing, as an active ingredient, a Val-Val-Try-Pro peptide isolated from a globin proteolysate.
For example, Patent Document 2 contains neutral fat mass adjustment characterized by containing at least one of protopanaxatriol, panaxatriol, protopanaxatriol, and panaxadiol, which is an aglycon form of dammaran saponin. Agents have been proposed.

JP-A-9-255698 JP 2011-12005 A

However, highly safe blood triglyceride lowering inhibitors, blood LDL / HDL cholesterol ratio lowering agents, and lipid metabolism improving agents are desired.
The present invention is intended to provide any one selected from a blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent, and a lipid metabolism improving agent.

As a result of intensive studies, the present inventors have found that casein degradation products have a blood triglyceride lowering action, a blood LDL / HDL cholesterol ratio lowering action, and a lipid metabolism improving action.
That is, the present invention includes a casein degradation product having a molecular weight of 337 Da as an active ingredient , and the casein degradation product is used at a dose of 100 mg / kg body weight / day, for reducing blood triglyceride and blood LDL / An agent for reducing the HDL cholesterol ratio or a food / beverage product composition is provided.
Further, the casein degradation product may be a hydrolysis product of a casein protein hydrolase .

  The present disclosure can provide any one selected from a blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent, and a lipid metabolism improving agent with high safety.

FIG. 1 shows blood triglycerides in four groups: a non-pathological group (normal control), a casein 100 mg / kg body weight administration group, a casein degradation product 25 mg / kg body weight administration group, and a casein degradation product 100 mg / kg body weight administration group. It is a figure which shows a density | concentration. FIG. 2 shows blood LDL in four groups: a non-pathological group (normal control), a casein 100 mg / kg body weight administration group, a casein degradation product 25 mg / kg body weight administration group, and a casein degradation product 100 mg / kg body weight administration group. It is a figure which shows / HDL concentration ratio.

  Preferred embodiments of the present disclosure will be described in detail. However, the present disclosure is not limited to the following preferred embodiments, and can be freely changed within the scope of the present disclosure.

  The casein hydrolyzate of the present disclosure is preferably a hydrolyzed casein. The casein decomposition treatment is not particularly limited, and examples thereof include enzyme treatment, acid treatment, alkali treatment, heat treatment, and the like, and two or more treatments may be appropriately combined.

The non-protein nitrogen ratio of the casein degradation product is preferably 5 to 40%, more preferably 10 to 30%.
<Calculation method of non-protein nitrogen ratio>
The total nitrogen content of the sample was measured according to the Kjeldahl method edited by the Japan Food Industry Association, “Food Analysis Method”, page 102, Korin Co., Ltd. In addition, a measurement kit (trade name: NPN-Test Wako; manufactured by Wako Pure Chemical Industries, Ltd.) based on Rappaport-Modified Umeda (Clinical Laboratory, Vol. 9, pp. 534 to 537, 1965) is used. Then, the amount of non-protein nitrogen of the sample was measured according to the instructions of the measurement kit, and the amount obtained was multiplied by 6.38 to calculate the amount of non-protein nitrogen compound. From these measured values, the non-protein nitrogen ratio (%) is calculated by the following formula.
Non-protein nitrogen ratio (%) = (non-protein nitrogen compound amount / total nitrogen amount) × 100

  The amino acid release rate of the casein degradation product is preferably 10% or less, more preferably 5% or less.

<Calculation method of amino acid release rate>
For amino acids other than tryptophan, cysteine and methionine, the sample was hydrolyzed with 6N hydrochloric acid at 110 ° C. for 24 hours, and for tryptophan, the sample was alkali-degraded with barium hydroxide at 110 ° C. for 22 hours. For cysteine and methionine, After treatment with performic acid, hydrolysis was performed with 6 N hydrochloric acid at 110 ° C. for 18 hours, and each was analyzed with an amino acid automatic analyzer (manufactured by Hitachi, Ltd. Model 835) to measure the mass of amino acids.
The composition of each amino acid in the sample is measured by the above method, and these are summed to calculate the mass of all amino acids in the sample. Next, the sample is deproteinized with sulfosalicylic acid, the mass of each free amino acid remaining is measured by the above method, and the total is calculated to calculate the mass of all free amino acids in the sample. From these values, the free amino acid content in the sample was calculated by the following equation.
Amino acid release rate (%) = (mass of all free amino acids / mass of all amino acids) × 100

  The molecular weight of the casein degradation product is preferably 2000 daltons (hereinafter referred to as “Da”) or less, and more preferably 1000 Da or less.

<Method for calculating molecular weight>
The molecular weight of the casein degradation product in the present disclosure is determined by the following concept of number average molecular weight.
The number average molecular weight (Number Average of Molecular Weight) is described in, for example, the literature (edited by the Society of Polymer Science, “Basics of Polymer Science”, pages 116 to 119, Tokyo Kagaku Dojin, 1978). As shown, the average value of the molecular weight of the polymer compound is shown based on different indicators as follows.
That is, high molecular weight compounds such as protein hydrolysates are heterogeneous substances and have a distribution in molecular weight. Therefore, the molecular weight of protein hydrolysates must be expressed as an average molecular weight in order to handle them physicochemically. The number average molecular weight (hereinafter sometimes abbreviated as Mn) is the average of the number of molecules, the molecular weight of the peptide chain i is Mi, and the number of molecules is Ni. Defined.

The casein as the starting material in the casein decomposition product is, for example, one kind selected from commercially available casein, lactic acid casein, acid casein such as casein hydrochloride; sodium caseinate, potassium caseinate, calcium caseinate, and the like. Or a mixture of two or more. What is necessary is just to mix the said mixture in arbitrary ratios. Moreover, it is preferable that the said casein is casein isolated by a well-known method from cow's milk, skim milk, whole milk powder, skim milk powder, etc.
That is, the casein decomposition product used in the present invention is preferably a hydrolysis product obtained by hydrolysis using casein, which is a protein, as a starting material.

  Raw material casein is dispersed in water or hot water and dissolved to prepare a casein aqueous solution. The concentration of the casein aqueous solution is not particularly limited, but it is usually preferable to set the protein concentration to a concentration range of 2% or more, and further about 5 to 15% by mass.

  Further, the casein aqueous solution may be obtained by ion exchange using sodium or potassium cation exchange resin (preferably strongly acidic cation exchange resin), electrodialysis, ultrafiltration membrane method, loose reverse osmosis membrane method, etc. It is preferable to desalinate and adjust pH and calcium concentration appropriately. In desalting, either a column type or a batch type may be employed. Moreover, you may heat-sterilize casein aqueous solution suitably before desalting etc.

  Next, the casein aqueous solution is hydrolyzed. Examples of the hydrolysis treatment include acid treatment, alkali treatment, heat treatment, enzyme treatment, and the like, and two or more treatments may be appropriately combined.

  Examples of the proteolytic enzyme of the present disclosure include plant-derived, animal-derived, and microorganism-derived, and can be used alone or in combination of two or more thereof. An endoprotease is suitable as the proteolytic enzyme.

Examples of the endoprotease include serine protease, metalloprotease, cysteine protease, and aspartic proteinase, and one or more of these can be selected and used. Among these, it is preferable to use serine protease and / or metalloprotease.
Proteases are classified into alkaline proteases, neutral proteases and acidic proteases. Of these, neutral protease is preferably used.

A commercially available product can be used as the proteolytic enzyme. Examples of the proteolytic enzyme include, for example, bioprase (manufactured by Nagase Seikagaku Corporation), pro leather (manufactured by Amano Enzyme), protease S (manufactured by Amano Enzyme), PTN 6.0S (manufactured by Novozymes), sabinase (Novozymes) Manufactured by GODO B. A. P (manufactured by Godo Shusei Co., Ltd.), protease N (manufactured by Amano Enzyme), GODO B. N. P (manufactured by Godo Shusei), Neutase (manufactured by Novozymes), Alcalase (manufactured by Novozymes), Trypsin (manufactured by Novozymes), Chymotrypsin (manufactured by Novozymes), Subtilisin (manufactured by Novozymes), Papain (manufactured by Amano Enzymes) ), Bromelain (manufactured by Amano Enzyme), etc., and one or more enzymes may be selected from these.
Among these, one or more selected from subtilisin (for example, bioplase), trypsin (for example, PTN6.0S), and basilosin (for example, protease N) are preferable, and these are neutral. It is a protease. Furthermore, a combination of these three types is preferable.

  The amount of endoprotease used for the casein is not particularly limited, and may be appropriately adjusted depending on the substrate concentration, enzyme titer, reaction temperature, reaction time, etc. Generally, 2000 g per 1 g of protein in casein. It is preferable to add at a rate of ˜11000 active units.

  The hydrolysis by the proteolytic enzyme is desirably performed so as to satisfy desired conditions (for example, within a molecular weight range). Thereby, the casein decomposition product used by this indication can be obtained.

  Prior to hydrolysis by the proteolytic enzyme, the pH of the raw milk protein solution can be adjusted to the optimum pH of the enzyme used with salts that can be used on food such as potassium carbonate and sodium hydroxide. The pH of the raw milk protein solution is preferably adjusted to 5 to 10, more preferably 7 to 10.

  The reaction temperature of the proteolytic enzyme is desirably within the optimum temperature range of the enzyme used, preferably 30 to 60 ° C, more preferably 40 to 60 ° C.

  The reaction holding time of the proteolytic enzyme may be appropriately adjusted so as to be the specific non-protein nitrogen ratio, and can be performed, for example, in 0.5 to 24 hours, preferably 1 to 15 hours, More preferably, it is 3 to 10 hours.

The hydrolysis by the proteolytic enzyme may be terminated by heating and deactivating the enzyme. For example, when deactivating at 100 ° C. or higher (preferably 110-130 ° C.) for 1 to 3 seconds, when deactivating at less than 100 ° C. and 60 ° C. or higher, it is preferable to perform for 3 to 40 minutes. .
After completion of hydrolysis, the pH of the decomposition solution is preferably 6 to 8, more preferably 7.0 ± 0.5, and even more preferably 7.0 ± 0.3, if necessary.

  In the production of the casein degradation product of the present disclosure, when the calcium concentration unadjusted solution is hydrolyzed, the obtained decomposition solution may be desalted as described above to adjust the calcium concentration. . Next, the enzyme is inactivated by heating by a conventional method. The reaction heating temperature and the reaction holding time can be set as appropriate under conditions that allow for sufficient deactivation in consideration of the thermal stability of the enzyme used. After heat deactivation, it can be cooled by a conventional method and used as it is, or it can be concentrated as necessary to obtain a concentrate, and the concentrate can be dried to obtain a powder product.

Moreover, what is necessary is just to adjust and adjust the pH of casein aqueous solution, when hydrolyzing the casein aqueous solution by acid treatment or alkali treatment. In the case of the treatment by pH adjustment, the pH of the casein aqueous solution is preferably pH 5 or lower or pH 9 or higher, more preferably pH 4 or lower or pH 10 or higher. The pH-treated aqueous solution can be left or stirred at room temperature for several minutes or more, preferably 5 minutes to 1 hour, to obtain an acid-treated or alkali-treated hydrolyzate. Here, “room temperature” is about 4 to 40 ° C., but 10 to 30 ° C. is preferable.
The casein aqueous solution may be hydrolyzed by heat treatment. This aqueous casein solution may be pH-unadjusted, or may be pH-adjusted (specifically, acidic (pH 5 or lower), neutral (pH 6-8), alkaline (pH 8 or higher)). The heat treatment may be performed at about 4 to 100 ° C. under the conditions such as the acid-alkali treatment.

The obtained casein hydrolyzate can exhibit efficacy even when used in an unpurified state. Furthermore, in order to improve efficacy, it is also possible to obtain a component having the efficacy of the present disclosure or a fraction having this component by performing known separation and purification as appropriate.
For example, molecular weight fractionation can be performed on the obtained casein hydrolyzate to enhance the efficacy of the present disclosure. Thereby, it is preferable to make the casein hydrolyzate less than or less than 2000 Da in molecular weight, and more preferably less than 1000 Da in molecular weight.

As the molecular weight fractionation, for example, methods such as ultrafiltration and gel filtration can be employed, whereby the removal rate of unnecessary molecular weight peptides can be increased.
In the case of ultrafiltration, a desired ultrafiltration membrane may be used, and in the case of gel filtration, a gel filtration agent used for desired size exclusion chromatography may be used.
Furthermore, a known separation and purification method (for example, an ion exchange resin or the like) may be used for desalting, removing impurities, and increasing the purity.

The casein degradation product of the present disclosure has a blood triglyceride concentration lowering action and a blood LDL / HDL cholesterol ratio lowering action, as shown in Examples below. Therefore, the casein degradation product of the present disclosure can be used as a blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent, and an increase in blood triglyceride concentration, blood LDL concentration / HDL cholesterol concentration. It can be used for control of various conditions related to an increase in the ratio, for example, prevention, improvement and / or treatment of diseases such as abnormal lipid metabolism, hyperlipidemia, hypertriglyceridemia and the like.
Therefore, the blood triglyceride lowering agent, blood LDL / HDL cholesterol ratio lowering agent, lipid metabolism abnormality improving agent, hypertriglyceridemia improving agent and the like of the present disclosure contain the casein degradation product of the present disclosure as an active ingredient.

The above-mentioned use may be a use in a human or non-human animal to be applied, and may be a therapeutic use or a non-therapeutic use.
In the present specification, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by therapy.
In the present specification, “improvement” refers to improvement of a disease, symptom or condition; prevention, delay or deterioration of progression of the disease, symptom or condition, reversal, prevention or delay of progression of the disease or symptom.
In the present specification, “prevention” means prevention or delay of the onset of the disease or symptom in the application target, or reduction of the risk of the onset of the disease or symptom of the application target.

  Accordingly, the casein degradation product of the present disclosure and the above-mentioned various preparations containing this as an active ingredient (hereinafter referred to as “blood TG lowering agents”) are ingested or administered to animals including humans, Methods for improving blood triglyceride, blood LDL / HDL cholesterol ratio, diseases related to lipid metabolism abnormalities, improvement of diseases and symptoms, or prevention of hyperlipidemia, hypertriglyceridemia, etc. It can be used in a method for improving and / or treating.

  Further, the casein degradation product of the present disclosure and the above-mentioned various preparations containing this as an active ingredient include the above-described rise in blood triglyceride, rise in blood LDL / HDL cholesterol ratio, diseases, diseases and diseases involving lipid metabolism abnormality, etc. For humans or veterinary drugs, quasi-drugs, topical skin preparations, cosmetics, foods, etc. for symptoms or for prevention, improvement and / or treatment of hyperlipidemia, hypertriglyceridemia, etc. It can be blended and used as an active ingredient. Moreover, the casein degradation product of the present disclosure can be used for the production of these various preparations.

When blended in a pharmaceutical product, it can be an orally administered agent or a parenterally administered agent.
In addition, when blended in foods, prevention, improvement of various diseases caused by the above-mentioned blood triglyceride elevation, blood LDL / HDL cholesterol ratio elevation, lipid metabolism abnormality, hyperlipidemia, hypertriglyceridemia, or It can be applied to functional foods with the concept of treatment, foods for the sick, foods for specified health use, etc. Moreover, the casein decomposition product of this indication can be used for manufacture of these foodstuffs.

The casein degradation product of the present disclosure and the above-mentioned various preparations containing this as an active ingredient may be either oral administration or parenteral administration, but oral administration is desirable. Parenteral administration includes intravenous injection, rectal administration, inhalation and the like. Examples of the dosage form for oral administration include tablets, capsules, troches, syrups, granules, powders and ointments.
In formulating, in addition to the casein degradation product of the present disclosure, components such as excipients, pH adjusters, colorants, and corrigents that are usually used for formulation can be used. It is also possible to use a known or future-discovered drug having an inhibitory action on the increase in blood triglyceride, a lipid metabolism abnormality improving drug, a hyperlipidemia drug, and the like.

In addition, the casein degradation product of the present disclosure is contained in a food as an active ingredient, and the casein degradation product of the present disclosure and the above-described various preparations containing this as an active ingredient include the above-described blood triglyceride increase inhibitory action and the like. It is also possible to process it as a food having
Examples of such foods include liquids, pastes, solids, powders, etc. In addition to tablet confectionery, liquid foods, feeds (including for pets), etc., for example, flour products, instant foods, processed agricultural products, Processed marine products, processed livestock products, milk / dairy products, fats and oils, basic seasonings, compound seasonings / foods, frozen foods, confectionery, beverages, commercial foods other than these, and the like.

For example, examples of the flour product include bread, macaroni, spaghetti, noodles, cake mix, fried flour, bread crumbs and the like.
Examples of the instant foods include instant noodles, cup noodles, retort / cooked food, cooking canned food, microwave food, instant soup / stew, instant miso soup / soup, canned soup, freeze-dried food, and other instant foods.
Examples of the processed agricultural products include canned agricultural products, canned fruits, jams and marmalades, pickles, boiled beans, dried agricultural products, cereals (cereal processed products), and the like.
Examples of the processed fishery products include canned fishery products, fish hams and sausages, fish paste products, marine delicacies, and tsukudani.
Examples of the processed livestock products include canned livestock, pastes, livestock ham, sausages and the like.

Examples of the milk / dairy products include processed milk, milk beverages, yogurts, lactic acid bacteria beverages, cheese, ice creams, prepared powdered milks, creams, and other dairy products.
Examples of the fats and oils include butter, margarines, and vegetable oils.
For example, examples of the basic seasoning include soy sauce, miso, sauces, processed tomato seasonings, mirin, and vinegar.
Examples of the complex seasonings and foods include cooking mixes, curry ingredients, sauces, dressings, noodle soups, spices, and other complex seasonings.
Examples of the frozen food include raw material frozen food, semi-cooked frozen food, cooked frozen food, and the like.

Examples of the confectionery include caramel, candy, chewing gum, chocolate, cookies, biscuits, cakes, pie, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, and other confectionery.
For example, the beverages include carbonated beverages, natural fruit juices, fruit juice beverages, soft drinks with fruit juices, fruit drinks, fruit drinks with fruits, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks Sports drinks, nutritional drinks, alcoholic drinks, other taste drinks, and the like.
For example, examples of commercially available foods other than the above include baby foods, sprinkles, and tea paste.

In the blood triglyceride lowering agent and lipid metabolism improving agent of the present disclosure, the content of the casein degradation product of the present disclosure is preferably at least 0.001% by mass with respect to the final composition of the preparation.
The intake or dose of the casein degradation product of the present disclosure varies depending on age, symptoms, etc., but is usually 0.001 to 3000 mg / kg body weight / day, preferably 0.01 to 200 mg / kg body weight / day, It may be administered once to three times a day. The intake or dose for humans is preferably 10 mg / kg body weight / day.

The dose at the time of administration of the casein degradation product to humans can be calculated from a conversion formula by “HED (Human Equivalent Dose) exchange from animals based on body surface area” (for example, see Reference 1).
HED = [dosage to animal (mg / kg body weight)] × {[animal body weight (kg)] ÷ [human body weight (kg)]} ^0.33
Human weight: 60kg
Mouse weight: 20g
Reference 1: Guidance for Industry, Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers, V. STEP 2: HUMAN EQUIVALENT DOSE CALCULATION, July 2005, Pharmacology and Toxicology, p.6-7 / US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER)

The present technology can also employ the following configurations.
[1] A blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent, or a lipid metabolism improving agent comprising a casein degradation product as an active ingredient. It can also be used as a therapeutic agent for hyperlipidemia or a therapeutic agent for hypertriglyceridemia.
[2] The blood triglyceride lowering agent, the blood LDL / HDL cholesterol ratio lowering agent, or the lipid metabolism improving agent according to [1], wherein the casein degradation product is a hydrolyzate by a protein hydrolase.
[3] The blood triglyceride lowering agent, the blood LDL / HDL cholesterol ratio lowering agent, or the lipid metabolism improving agent according to [1] or [2], wherein the casein degradation product has a molecular weight of 1000 Da or less.
[4] The blood triglyceride reducing agent, blood LDL / HDL cholesterol ratio reducing agent or lipid metabolism improving agent according to [2] or [3], wherein the protein hydrolase is an endonuclease.

[5] Use of a casein degradation product for the production of a blood triglyceride lowering agent, a blood LDL / HDL cholesterol ratio lowering agent or a lipid metabolism improving agent.
[6] Use of a casein degradation product for the production of a food for lowering blood triglyceride, a food for lowering blood LDL / HDL cholesterol ratio or a food for improving lipid metabolism.
[7] Casein degradation product, blood triglyceride lowering agent, blood LDL / HDL cholesterol ratio lowering agent or lipid metabolism improving agent, blood triglyceride lowering food, blood LDL / HDL cholesterol ratio lowering food or lipid metabolism Use in foods for improvers.

[8] For prevention, improvement or treatment of blood triglyceride rise, blood LDL / HDL cholesterol ratio rise, disease, disease or symptom related to abnormal lipid metabolism, hyperlipidemia, hypertriglyceridemia, etc. Casein degradation product.
[9] Use in prevention, amelioration, or treatment of diseases, diseases or symptoms associated with elevated blood triglyceride, elevated blood LDL / HDL cholesterol ratio, abnormal lipid metabolism, etc., or hyperlipidemia, hypertriglyceridemia, etc. For casein degradation product.
[10] For prevention, improvement or treatment of blood triglyceride elevation, blood LDL / HDL cholesterol ratio elevation, disease, disease or symptom involving lipid metabolism abnormality, or hyperlipidemia, hypertriglyceridemia, etc. Use of casein degradation products.
[11] Ingestion or administration of casein degradation product as an active ingredient, blood triglyceride elevation, blood LDL / HDL cholesterol ratio elevation, illness or symptom involving lipid metabolism abnormality, hyperlipidemia, high triglyceride How to prevent, improve, or treat blood.

[12] The casein degradation product according to any one of [5] to [11] is a degradation product resulting from one or more treatments selected from heat treatment, acid treatment, alkali treatment, and proteolytic enzyme treatment. Is preferred.
[13] The casein degradation product according to [12] is preferably a hydrolysis product of casein by a protein hydrolase.
[14] The casein degradation product according to any one of [5] to [13] preferably has a molecular weight of 1000 Da or less.

  Specific examples and the like will be described below, but the present invention (the present disclosure) is not limited thereto.

<Example: Lipid metabolism-related test of casein enzyme degradation product>
[Production Example 1: Production of peptide by enzymatic degradation of casein]
Add 900 mg of water to 100 mg of commercially available casein (manufactured by New Zealand Daily Board), disperse well, adjust the pH of the solution to 7.0 by adding sodium hydroxide, completely dissolve the casein, concentration of about 10% An aqueous casein solution was prepared.
The aqueous casein solution is sterilized by heating at 85 ° C for 10 minutes, adjusted to 50 ° C, adjusted to pH 9.5 by adding sodium hydroxide, and then 100 units of biolase sp-20 (manufactured by Nagase Seikagaku Corporation) (1200 active units per gram of protein), protease N (manufactured by Amano Enzyme) 170 active units (2000 active units per gram of protein), and PTN6.0S (manufactured by Novozymes Japan) 600 active units (7000 active units per gram of protein) ) Was added to initiate the hydrolysis reaction. After 8 hours, the enzyme reaction was stopped by heating at 80 ° C. for 6 minutes to stop the enzyme reaction and cooled to 10 ° C.
This hydrolyzed solution was ultrafiltered with an ultrafiltration membrane (manufactured by Nippon Pall Co., Ltd.) having a molecular weight cut off of 1000, concentrated and freeze-dried to obtain 85 mg of a freeze-dried product.

  The casein hydrolyzate of Production Example 1 had a non-protein nitrogen ratio of 25%, an amino acid release rate of 5%, and a molecular weight of 337 Da. These were calculated by the above <Method for calculating non-protein nitrogen ratio>, <Method for calculating amino acid release rate>, and <Method for calculating molecular weight>.

〔experimental method〕
Experimental animal: B6.KOR / Stm Slc-Apoe ^shl (Apo E deficient mouse: Atherosclerosis model, Japan SLC), B6 (C57BL / 6J) mouse as non-pathological animal.
Experimental method: MF feed (Oriental Yeast Co., Ltd.) supplemented with 1.25% cholesterol and 10% coconut oil was ingested as soon as the experimental animals arrived at the age of 6 weeks. The non-pathological group had free intake of MF diet.
After acclimation for 1 week, casein 100 mg / kg body weight group (undegraded casein: casein 100 mg / kg), casein degradation product 25 mg / kg body weight group (casein hydrolysate 25 mg / kg), and casein The degradation product was divided into three groups of 100 mg / kg body weight (casein hydrolysate 100 mg / kg), and each was orally administered once a day for 8 weeks using an oral sonde.
After the final administration, the experimental animals were fasted for 16 hours, and then blood was collected from the inferior vena cava under isoflurane anesthesia and centrifuged at 12000 rpm for 15 minutes to collect plasma. The total triglyceride concentration, total cholesterol concentration, and HDL cholesterol concentration in blood were measured using Spot Chem II and Screening-2 (Arkray).
The LDL cholesterol concentration was calculated from the formula [total cholesterol concentration] − [HDL cholesterol concentration] − [neutral fat concentration] ÷ 5.

〔result〕
In animal experiments using an arteriosclerosis model, oral administration of a casein degradation product has an effect of suppressing an increase in blood triglyceride concentration compared to undegraded casein, and the dose is 100 mg / kg body weight. The concentration difference was significant. The blood LDL / HDL cholesterol concentration ratio also showed a similar tendency (see FIGS. 1 and 2).
Casein enzyme hydrolyzate has the effect of suppressing blood triglyceride concentration increase and blood LDL / HDL cholesterol ratio decreasing action, so it can be used for prevention, improvement or treatment of lipid metabolism abnormality, or prevention, improvement or treatment of dyslipidemia can do.

Claims (4)

The molecular weight is contained as an active ingredient casein hydrolyzate is 337Da, the casein hydrolyzate is 100 mg / kg body weight / day, wherein the to Ruchichu triglyceride for lowering and blood LDL / HDL-cholesterol ratio for lowering to be used in Agent. The agent according to claim 1 , wherein the casein degradation product is a hydrolysis product of a protein hydrolase. A casein degradation product having a molecular weight of 337 Da is contained as an active ingredient, and the casein degradation product is used at a dose of 100 mg / kg body weight / day, for blood triglyceride reduction and blood LDL / HDL cholesterol ratio reduction Food and beverage composition. The food / beverage product composition according to claim 3, wherein the casein degradation product is a hydrolysis product of a protein hydrolase.

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