JP2008156344A - In vivo antioxidant, food composition for in vivo antioxidation, pharmaceutical composition for in vivo antioxidation, and inhibitor for liver function disturbance, food composition for inhibition of liver function disturbance and pharmaceutical composition for inhibition of liver function disturbance - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an in vivo antioxidant, a food composition for in vivo antioxidation and a pharmaceutical composition for in vivo antioxidation, and an inhibitor for liver function disturbance, a food composition for the inhibition of liver function disturbance and a pharmaceutical composition for inhibition of liver function disturbance, which can be commercially produced with ease at a low cost and have a high practical use. <P>SOLUTION: The in vivo antioxidant contains, as an effective ingredient, an egg white hydrolyzate prepared by the treatment with a neutral protease derived from genus Aspergillus fungi, and the inhibitor for liver function disturbance contains the in vivo antioxidant. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

The present invention relates to an in vivo antioxidant material, an in vivo antioxidant food composition, an in vivo antioxidant pharmaceutical composition, a liver dysfunction inhibitor, a liver dysfunction food composition, and a liver dysfunction drug. Relates to the composition.

Conventionally, in vivo oxidation by active oxygen is said to cause various diseases such as cancer, arteriosclerosis, inflammation, complications due to diabetes, liver dysfunction, and Alzheimer's disease. However, in vivo oxidation cannot escape as long as you are breathing. Accordingly, the appearance of foods and pharmaceuticals that remove active oxygen generated in vivo and suppress in vivo oxidation is desired.

As a substance exhibiting in vivo antioxidant action, for example, a reactive oxygen inhibitor of neutrophils having a peptide represented by the amino acid sequence of His-Ser-Ser-Leu-Arg (Patent Document 1), or derived from milk protein An antioxidant food or drink or feed containing a peptide comprising a specific amino acid sequence (Patent Document 2) is disclosed. However, all of the substances described in these patent documents use a peptide having a specific amino acid sequence, and it is difficult to industrially produce a peptide having a specific amino acid sequence. It has not reached.

JP 2000-143696 A JP 2006-131626 A

The object of the present invention is industrially easy to produce, inexpensive and highly practical in vivo antioxidant material, in vivo antioxidant food composition and in vivo antioxidant pharmaceutical composition, and liver dysfunction inhibitor, To provide a food composition for suppressing liver dysfunction and a pharmaceutical composition for suppressing liver dysfunction.

In order to achieve the above object, the present inventors have conducted extensive research on antioxidative substances, and as a result, surprisingly, egg white hydrolyzate having antioxidative properties to foods also has an antioxidant effect in vivo. As a result, the present invention has been completed.

That is, the present invention
(1) An in vivo antioxidant containing egg white hydrolyzate as an active ingredient,
(2) The in vivo antioxidant material according to (1), wherein the egg white hydrolyzate is treated with a neutral protease derived from Aspergillus,
(3) The in vivo antioxidant material according to (2), wherein the egg white hydrolyzate is further treated with papain.
(4) The in vivo antioxidant material according to any one of (1) to (3), wherein the in vivo antioxidant material is a lipid antioxidant material of a biological membrane,
(5) The in vivo antioxidant material according to (4), wherein the lipid antioxidant of the biological membrane is a HEL production inhibitor,
(6) The in vivo antioxidant material according to (4) or (5), wherein the lipid antioxidant material of the biological membrane is a lipid antioxidant material of hepatocytes,
(7) In vivo antioxidant food composition comprising the in vivo antioxidant material according to any one of (1) to (6),
(8) In vivo antioxidant pharmaceutical composition comprising the in vivo antioxidant material according to any one of (1) to (6),
(9) A liver dysfunction inhibitor comprising the in vivo antioxidant material according to (6) as an active ingredient,
(10) A food composition for inhibiting liver dysfunction comprising the liver dysfunction inhibiting material according to (9),
(11) A pharmaceutical composition for inhibiting liver dysfunction comprising the liver dysfunction inhibiting material according to (9),
It is.

INDUSTRIAL APPLICABILITY The present invention can provide an in-vivo antioxidant that is easy to industrially produce, contains inexpensive and highly practical egg white hydrolyzate as an active ingredient. In addition, by using the in vivo antioxidant material of the present invention in a food composition or a pharmaceutical composition, it is possible to prevent or improve various diseases such as cancer, arteriosclerosis, inflammation, complications due to diabetes, liver dysfunction, and Alzheimer's disease. In-vivo antioxidant food composition and in-vivo antioxidant pharmaceutical composition are obtained. Further, the present invention can provide a liver dysfunction inhibiting material containing the in vivo antioxidant as an active ingredient, and the liver dysfunction inhibiting material can be used for foods and pharmaceuticals, thereby preventing liver dysfunction. Alternatively, a food composition for suppressing hepatic dysfunction and a pharmaceutical composition for suppressing hepatic dysfunction can be obtained.

The present invention will be described below. In the present invention, “%” means “% by mass” unless otherwise specified.

The in-vivo antioxidant of the present invention contains egg white hydrolyzate as an active ingredient. That is, the egg white hydrolyzate used in the present invention has an in vivo antioxidant action that suppresses in vivo oxidation caused by active oxygen, and thus can be used as an in vivo antioxidant material.

In addition, among the in vivo oxidation caused by active oxygen, the egg white hydrolyzate used in the present invention has an action of suppressing the formation of lipid peroxide in the biological membrane, so it can be used as a lipid antioxidant for biological membranes. In particular, the egg white hydrolyzate has an action of suppressing the production of HEL (hexanoyl lysine) generated in the initial stage of the production of lipid peroxide in such a biological membrane. It can be used. Furthermore, since the egg white hydrolyzate used in the present invention has an action of suppressing the production of lipid peroxide in the hepatocyte biomembrane, it can be used as a lipid antioxidant for hepatocytes.

By using such an in vivo antioxidant material of the present invention for foods and pharmaceuticals, various diseases such as liver dysfunction caused by in vivo oxidation, cancer, arteriosclerosis, inflammation, diabetes complications, Alzheimer's disease, etc. In vivo antioxidant food composition and in vivo antioxidant pharmaceutical composition having an effect of preventing or improving the above can be obtained.

Moreover, the liver dysfunction inhibitor of this invention contains the said in-vivo antioxidant, ie, an egg white hydrolyzate, as an active ingredient. Among the above diseases, hepatic dysfunction is caused by the production of lipid peroxide in the hepatocyte biomembrane, and the antioxidant of the present invention has the action of suppressing the production of lipid peroxide in the hepatocyte biomembrane. Therefore, it can be used as a liver dysfunction inhibitor.

By using the liver dysfunction-suppressing material of the present invention in foods and pharmaceuticals, a food composition for suppressing liver dysfunction and a pharmaceutical composition for suppressing liver dysfunction having an effect of preventing or improving liver dysfunction can be obtained.

The egg white hydrolyzate used in the present invention is a hydrolyzed egg white, and may be used as a raw material for the in vivo antioxidant of the present invention in the form of an aqueous suspension after hydrolysis, or After drying, it may be used as a raw material for the in vivo antioxidant material of the present invention. Examples of the method for drying the egg white hydrolyzate include spray drying and freeze drying.

In the present invention, the egg white used in producing the egg white hydrolyzate is, for example, raw egg white, frozen egg white, dried egg white rehydrated, egg white from which a specific egg white protein (such as lysozyme) has been removed, etc. Is mentioned. Here, the raw egg white means one obtained by dividing a chicken egg or the like and separating the egg yolk. Moreover, the egg white may have been subjected to a desugaring treatment with yeast, bacteria or enzymes, for example.

In the present invention, the egg white hydrolysis method may be either chemical treatment or enzyme treatment. The chemical treatment includes decomposition with acid, alkali, methyl bromide and the like. From the viewpoint of quality stability, hydrolysis by enzyme treatment is preferred, and enzyme treatment by protease is particularly preferred.

In the present invention, examples of proteases used in producing egg white hydrolyzate include animal origin (eg, pepsin, chymotrypsin, trypsin, pancreatin), plant origin (eg, papain, bromelain, ficin), microorganisms Examples include endoproteases and exoproteases of origin (for example, lactic acid bacteria, Bacillus subtilis, actinomycetes, fungi, yeast), and crudely purified products and crushed cells thereof, which are used alone or in combination of two or more. be able to. Among these, in order to obtain an egg white hydrolyzate having an excellent in vivo antioxidant effect, it is preferable to hydrolyze the egg white using a neutral protease derived from the genus Aspergillus as a protease.

As the neutral protease derived from the genus Aspergillus, commercially available products can be used. For example, trade name: Protease P “Amano” 3G (origin: Aspergillus melleus, Amano Enzyme Co., Ltd.), trade name: Sumiteam FP (Origin: Aspergillus oryzae, Shin Nippon Chemical Industry Co., Ltd.), Trade name: Denateam AP (Origin: Aspergillus
oryzae, Nagase ChemteX Corporation).

For example, when the egg white is hydrolyzed using a neutral protease derived from Aspergillus, the pH of the egg white is adjusted to 6.5 to 9.5 (preferably pH 7), and the egg white is derived from Aspergillus. Neutral protease is added and the solution is held at 35-60 ° C (preferably 45-55 ° C) for 2-24 hours with slow stirring. Next, this liquid is heat sterilized, then cooled, and then a powdered egg white hydrolyzate can be obtained by spray drying or the like. In addition, pH, temperature conditions, and heating time may be appropriately adjusted according to the type and combination of proteases used.

In the present invention, the egg white hydrolyzate is more preferably obtained by hydrolyzing egg white using the above-mentioned neutral protease derived from the genus Aspergillus and further using papain. Papain is a protease extracted from the milk of papaya (Carica Papaya L). By hydrolyzing egg white using papain, hydrolysis can be promoted, and an egg white hydrolyzate having a superior in vivo antioxidant action can be obtained. In addition, when the egg white is hydrolyzed using only papain, the bitter taste of the obtained egg white hydrolyzate becomes strong, and when this egg white hydrolyzate is added to the food composition and the pharmaceutical composition, the food composition and the pharmaceutical composition of the present invention The flavor of things may be reduced.

Here, the order in which neutral protease and papain derived from Aspergillus are added to egg white is not particularly limited. Papain may be added first, or neutral protease derived from Aspergillus may be added first. Alternatively, neutral protease and papain from Aspergillus may be added simultaneously. Alternatively, hydrolysis may be performed using neutral protease and papain derived from Aspergillus.

As papain, commercially available products can be used. For example, trade name: purified papain for food (Nagase ChemteX Corporation), trade name: Papain F (Higuchi Shokai Co., Ltd.), trade name: Papain W -40 (Amano Enzyme Co., Ltd.), trade name: Papain (Solvay Enzymes, Inc.).

The food composition for in vivo antioxidants of the present invention contains an in vivo antioxidant material. That is, the food composition for in vivo antioxidant of the present invention contains egg white hydrolyzate which is an active ingredient of the in vivo antioxidant.

Moreover, the food composition for suppressing liver dysfunction of the present invention contains a liver dysfunction inhibitor. That is, the food composition for suppressing liver dysfunction according to the present invention includes an in vivo antioxidant that is an active ingredient of a liver dysfunction inhibitor.

The form of the egg white hydrolyzate to be added to the food composition of the present invention can be selected depending on the application. The blending amount of the in vivo antioxidant material of the present invention may be adjusted as appropriate depending on the food used, but if it is too small, it is difficult to obtain the in vivo antioxidant effect or the liver dysfunction inhibiting effect, The content of egg white hydrolyzate, which is an active ingredient of the in-vivo antioxidant in the food composition, is preferably 0.01 to 30% by mass because the effect may be difficult to obtain even if it is too much. More preferably, it is 0.05-10 mass%, More preferably, it is 0.3-5 mass%.

Although the aspect of the food composition of this invention is not specifically limited, For example, it can be processed food. Processed foods include, for example, confectionery, legume preparations, mayonnaise, dressings, dairy products (dairy beverages, fermented milk, lactic acid bacteria beverages, dairy lactic acid bacteria beverages, etc.), processed egg products, cooked foods, beverages (soft drinks) Etc.) and concentrated liquid foods.

Moreover, the food formulation (supplement) containing the in-vivo antioxidant of this invention can be prepared. In this case, by adding various food materials or beverage materials, for example, a food preparation in a powder form, a tablet form, a capsule form, a liquid form (such as a drink) can be obtained. Moreover, you may add a base material, an excipient | filler, an additive, a subsidiary material, a bulking agent, etc. to this food formulation suitably.

The food composition of the present invention can also be used in combination with other substances having a biological antioxidant action such as polyphenol. Moreover, it can also be used in combination with a substance having an effect of improving liver function, such as oyster extract.

The in vivo antioxidant pharmaceutical composition of the present invention contains an in vivo antioxidant material. That is, the in vivo antioxidant pharmaceutical composition of the present invention contains egg white hydrolyzate which is an active ingredient of the in vivo antioxidant.

The pharmaceutical composition for suppressing liver dysfunction of the present invention contains a liver dysfunction inhibitor. That is, the pharmaceutical composition for suppressing liver dysfunction of the present invention includes an in vivo antioxidant that is an active ingredient of a liver dysfunction inhibitor.

The form of the egg white hydrolyzate added to these pharmaceutical compositions of the present invention can be selected according to the application. The blending amount of the in vivo antioxidant material of the present invention may be appropriately adjusted depending on the pharmaceutical used, but if it is too small, it is difficult to obtain the in vivo antioxidant effect or the liver dysfunction inhibiting effect, The content of egg white hydrolyzate, which is an active ingredient of the in vivo antioxidant in the pharmaceutical composition, is preferably 0.01 to 30% by mass because the effect may be difficult to obtain even if it is too much. More preferably, it is 0.05-10 mass%, More preferably, it is 0.3-5 mass%.

The administration form of the pharmaceutical composition of the present invention is not particularly limited, and examples thereof include oral administration and tube administration. The administration time may be, for example, before a meal, after a meal, or between meals. Moreover, the frequency | count of administration is 1 to several times a day, for example.

Although the dosage form of the pharmaceutical composition of the present invention is not particularly limited, it can be an oral preparation. For example, the material containing the above-described in vivo antioxidant of the present invention can be processed into the following dosage form.

Examples of oral preparations include internal preparations such as tablets, capsules, powders, fine granules, granules, solutions, syrups, and drinks. In this case, the pharmaceutical composition of the present invention can contain a binder, an excipient, a swelling agent, a lubricant, a sweetener, a flavoring agent and the like together with the in-vivo antioxidant of the present invention. Here, the tablets can also be coated with shellac or sugar. In addition, the capsule may further contain a liquid carrier such as fats and oils in the above material. Syrups and drinks may contain sweeteners, preservatives, pigment flavors and the like.

Next, the present invention will be described more specifically with reference to examples. In addition, the following description shows the aspect of this invention generally, and this invention is not limited by this description without a particular reason.

Example 1
180 kg of liquid egg white (manufactured by QP Corporation) was put into a tank and adjusted to pH 7 using citric acid. Next, 300 g of yeast (manufactured by Oriental Yeast Co., Ltd.) was added, the liquid temperature was kept at 35 ° C., and desugaring treatment was performed for 4 hours with slow stirring. Thereafter, the liquid temperature was raised to 65 ° C., and the mixture was stirred at 65 ° C. for 30 minutes. Next, 150 g of neutral protease (trade name “Denateam AP”, manufactured by Nagase ChemteX Co., Ltd.) and papain (trade names “purified papain for food”, Nagase ChemteX ( 150 g) was added, the liquid temperature was kept at 55 ° C., and the enzyme treatment was performed for 6 hours with slow stirring. Further, the obtained enzyme treatment solution was treated with a kneader at 97 ° C. for 10 minutes, then cooled to 10 ° C. or less and spray-dried. As a result, an in-vivo antioxidant material of white powder composed of 13.7 kg of egg white hydrolyzate was obtained. The degree of decomposition of the egg white hydrolyzate by formol titration was 11.3.

Example 2
180 kg of liquid egg white (manufactured by QP Corporation) was put into a tank and adjusted to pH 7 using citric acid. Next, 200 g of a neutral protease (trade name “Sumiteam FP”, manufactured by Shin Nippon Chemical Industry Co., Ltd.) from Aspergillus oryzae is added to the liquid egg white, and the liquid temperature is kept at 45 ° C. while slowly stirring. Enzyme treatment was performed for 8 hours. Next, the obtained enzyme treatment liquid was treated with a kneader at a liquid temperature of 97 ° C. for 10 minutes, cooled to 10 ° C. or lower, and spray-dried. As a result, a white powder in-vivo antioxidant comprising 15.7 kg egg white hydrolyzate was obtained. The degree of decomposition of the egg white hydrolyzate by formol titration was 9.9.

Example 3
180 kg of liquid egg white (manufactured by QP Corporation) was put into a tank and adjusted to pH 7 using citric acid. Next, 300 g of yeast (manufactured by Oriental Yeast Co., Ltd.) was added, the liquid temperature was kept at 35 ° C., and desugaring treatment was performed for 4 hours with slow stirring. Next, 300 g of neutral protease (trade name “Protease P (Amano)”, manufactured by Amano Enzyme Co., Ltd.) derived from Aspergillus melleus was added to the resulting desugared solution, and the liquid temperature was maintained at 50 ° C. The enzyme treatment was performed for 12 hours with slow stirring. Further, the obtained enzyme treatment solution was treated with a kneader at 97 ° C. for 10 minutes, then cooled to 10 ° C. or less and spray-dried. As a result, a white powder in-vivo antioxidant comprising 16.3 kg of egg white hydrolyzate was obtained. The degree of decomposition of the egg white hydrolyzate by formol titration was 10.9.

Test Example 1 (Evaluation of In Vivo Antioxidant Action of In Vivo Antioxidant Containing Egg White Hydrolyzate) Next, a white powder in vivo antioxidant material consisting of egg white hydrolyzate obtained in Example 1 was used as an animal. It used for the sample of experiment and confirmed the in vivo antioxidant effect. The test methods and test results are shown below.

Test animals
System: Wistar Rat (SPF)
・
Source: Nippon SLC Co., Ltd.
・
Age (sex): 5 weeks of age (male) at the start of the study

Test sample The white powder in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1 was used.

The feed for which the composition of AIN-93G was modified was used. Among the compositions of AIN-93G, tert-butylhydroquinone and L-cystine, which are antioxidants, are excluded because they may suppress the enhancement of in vivo oxidation by CCl ₄ , and the vitamin mixture is vitamin E (V. E) No additives, oil is V.V. E-free added tryptocorn oil was used. Overall adjustment was made with casein and β-corn starch.

・
Each group was tested with n = 8.
・
CCl ₄ (special grade, Junsei Kagaku Co., Ltd.) was dissolved in tryptocorn oil so as to be 25% (v / v), and then intraperitoneally administered 2.0 ml / kg body weight (BW).

Test Contents After the rats were brought in, they were individually housed in a wire mesh cage and acclimated to the environment for 7 days. During the acclimatization period, the acclimatized food was freely ingested as feed. After acclimation, the modified AIN-93G containing no in vivo antioxidant (1%) diet in the CCl ₄ + in vivo antioxidant (1%) group, and no in vivo antioxidant in the control and CCl ₄ groups Each of the control meals, which are feeds of the composition, was freely ingested for 7 days.

Fasted overnight (20 o'clock to 10 o'clock the next day), CCl ₄ dissolved in tryptocorn oil was intraperitoneally administered, and fasting was continued until the end of the test. Twenty- _four hours after administration of CCl ₄ , blood was collected from the abdominal aorta under anesthesia with pentobarbital sodium (“Nenbutal” Dainippon Sumitomo Pharma Co., Ltd.), and the liver was collected after perfusion with physiological saline. After the obtained blood was coagulated at room temperature, serum was collected by centrifugation at 1000 × g for 15 minutes and used for serum HEL analysis. The collected liver was homogenized and centrifuged at 10,000 × g for 15 minutes, and the supernatant was used for liver TBARS and liver GSH analysis.

Evaluation item / Serum HEL: HEL is a substance specifically generated in the first stage of lipid peroxidation, and an increase in HEL serves as an index indicating that the living body is oxidized.
-Liver TBARS: TBARS is an aggregate of aldehydes such as malondialdehyde and alkenal produced in the second stage of lipid peroxidation, and an increase in TBARS serves as an index indicating that the living body is oxidized.
-Liver GSH: GSH is a substance consumed by glutathione peroxidase that scavenges active oxygen in the body, and a decrease in GSH is an indicator that active oxygen increases in the body and that the body is oxidized .

The serum HEL level was measured using a trade name: “ELISA kit for measuring hexanoyllysine” (Nikken Zile Co., Ltd.).

Liver TBARS values were measured with reference to the thiobarbituric acid (TBA) method. That is, in a 12 mL screw-cap test tube containing 0.1 mL of the supernatant obtained by homogenizing the liver, 8.1% (w / v) sodium dodecyl sulfate / 0.2 mL of distilled water, 0.8% (w / V) Butyl hydroxytoluene / acetic acid 0.05 mL, 0.8% (w / v) thiobarbituric acid / distilled water 1.5 mL, distilled water 0.7 mL Then, 1.5 mL of 20% (w / v) acetate buffer (pH 3.5) was added. The cap was capped and stirred, left at 5 ° C. for 1 hour, and then allowed to react at 100 ° C. for 1 hour. After cooling with running water, 1 mL of distilled water and 5 mL of butanol / pyridine (15/1) mixture were added, the cap was closed, the mixture was stirred, centrifuged at 10,000 × g for 15 minutes, and the absorbance of the supernatant (wavelength: 532 nm) ) Was measured. 110 mg of 1,1,3,3-tetramethoxypropane (1,1,3,3-tetraethoxypropane) was made up to 50 mL with 1% (v / v) sulfuric acid (H ₂ SO ₄ ), and the room temperature was 2 hours. Then, 1 mL was collected, and the volume up to 100 mL with distilled water was used as a standard for TBARS 100 nmol / L for preparing a calibration curve. The TBARS value (nmol / mL) quantified by the above method was converted to the amount (nmol / mg protein) per protein amount (mg / mL) measured by “Protein Assay Rapid Kit Wako”.

The liver GSH value was analyzed for the supernatant obtained by homogenizing the liver using a trade name: “Total Glutathione Quantification Kit” (Dojindo Laboratories).

The test results are shown as mean ± standard error, and a risk rate of less than 5% is considered significant. The significant difference test was performed by a one-way analysis of variance. When a significant difference was observed, analysis was performed by Fisher's PLSD test. When significant differences were observed, they were indicated by different alphabets in the tables and figures. In addition, computer software “Dr. SPSS II for Windows (registered trademark) (trade name)” (SPS Co., Ltd.) was used for statistical analysis.

Test results Serum HEL values are shown in Table 3. Serum HEL levels were significantly increased by CCl ₄ administration. In the CCl ₄ + in vivo antioxidant material (1%) group, the increase in the HEL value due to CCl ₄ administration was significantly suppressed.

Table 4 shows liver TBARS values. CCl ₄ administration significantly increased liver TBARS values. In the CCl ₄ + in vivo antioxidant material (1%) group, the increase in TBARS value due to CCl ₄ administration was significantly suppressed.

Table 5 shows liver GSH values. The CSH ₄ administration significantly decreased the GSH value. There was no significant effect on the GSH value due to the addition of the in vivo antioxidant.

From the above results, the increase in lipid peroxide, that is, serum HEL and liver TBARS due to carbon tetrachloride (CCl ₄ ) was suppressed by ingestion of the in vivo antioxidant (1%) diet. In-vivo antioxidants have been shown to have in vivo antioxidant effects. Serum HEL and liver TBARS are lipid peroxides called primary oxidation products and secondary oxidation products, respectively, and egg white hydrolyzate results in suppression of both of these two levels of lipid peroxides. I can understand that. In addition, since both serum HEL and liver TBARS are indices for measuring the degree of oxidation occurring in the living body hydrophobic part, that is, the biological membrane, egg white hydrolyzate suppresses the formation of lipid peroxide in the biological membrane. It can be said that it has been shown to have an action.

On the other hand, since the decrease in liver GSH by CCl ₄ was not suppressed by ingestion of the in vivo antioxidant (1%) diet, the in vivo antioxidant was not involved in the mechanism through liver GSH production. Conceivable. Moreover, when the test sample of Test Example 1 was changed to a white powdered in-vivo antioxidant made of egg white hydrolyzate obtained in Examples 1 to 2 and 3, the same test was performed. Although it was slightly inferior to Example 1, it was confirmed that an in vivo antioxidant effect was obtained.

Test Example 2 (Evaluation of liver dysfunction inhibiting action of liver dysfunction inhibitor made of egg white hydrolyzate)
The white powder in vivo antioxidant material consisting of the egg white hydrolyzate obtained in Example 1 was applied to a sample for animal experiments, and the liver function disorder inhibitory action was confirmed. The test methods and test results are shown below.

Test animals
System: ddY Mouse (SPF)
・
Supply source: Nippon SLC Co., Ltd.
・
Age (sex): 6 weeks of age (male) at the start of the study

・
Each group was tested with n = 4.
・
CCl ₄ (special grade, Junsei Kagaku Co., Ltd.) was dissolved in mineral oil so as to be 0.003% (v / v), and then 10.0 ml / kg body weight (BW) was intraperitoneally administered.

In Test Example 1, the mice were acclimated and bred for 7 days in the same manner as in Test Example 1, except that the test animals were changed to the aforementioned mice and the test group was changed to the test group shown in Table 6. Thereafter, the mouse liver and serum were collected in the same manner as in Test Example 1 except that the fasting was changed to a fasting for 4 hours. The following items were measured using the obtained liver and serum.

Evaluation item, liver TBARS
Serum GOT and GPT: GOT (glutamate oxaloacetate transaminase) and GPT (glutamate pyruvate transaminase) are enzymes that exist in the liver. When hepatocytes are damaged, they leak into the blood. An increase in GOT value or GPT value is an index indicating that hepatocytes are damaged by oxidation or the like.

Liver TBARS was measured in the same manner as in Test Example 1. Serum GOT and GPT values were measured using a trade name: “Transaminase C2-Test Wako” (manufactured by Wako Pure Chemical Industries, Ltd.).

Test results Table 7 shows liver TBARS values. CCl ₄ administration significantly increased liver TBARS values. In the CCl ₄ + in vivo antioxidant material (1%) group, the increase in TBARS value due to CCl ₄ administration was significantly suppressed.

Table 8 shows serum GOT values. Serum GOT levels were significantly increased by CCl ₄ administration. In the CCl ₄ + in vivo antioxidant material (1%) group, an increase in serum GOT value due to CCl ₄ administration was significantly suppressed.

Table 9 shows serum GPT values. Serum GPT levels were significantly increased by CCl ₄ administration. In the CCl ₄ + in vivo antioxidant material (1%) group, the increase in serum GPT value due to CCl ₄ administration was significantly suppressed.

From the above results, the intake of in vivo antioxidant (1%) diet suppresses the increase in lipid peroxide, that is, liver TBARS caused by carbon tetrachloride (CCl ₄ ), and is an index of the degree of liver dysfunction. Since the rise of certain serum GOT and GPT was suppressed, it was shown that the liver dysfunction inhibitor made of an in vivo antioxidant material (egg white hydrolyzate) has a liver dysfunction inhibitory action. Since the generation of liver TBARS, a lipid peroxide generated in hepatocyte biological membranes, is suppressed by carbon tetrachloride, egg white hydrolyzate has the effect of suppressing the production of lipid peroxides in hepatocyte biological membranes. Thus, it can be understood that liver dysfunction is suppressed. Moreover, when the test sample of Test Example 1 was changed to the white powder liver dysfunction inhibitor made of egg white hydrolyzate obtained in Examples 1 to 2 and 3, the same test was performed. Although it was slightly inferior to Example 1, it was confirmed that an effect of suppressing liver dysfunction was obtained.

Test example 3
Livers were collected from mice in the same manner as in Test Example 2 except that the amount of egg white hydrolyzate blended in the ingested feed in Test Example 2 was changed to the amount shown in Table 10. The liver TBARS value was measured using the obtained liver. The results are shown in Table 11.

Since the liver TBARS value is an index showing an in vivo antioxidant effect and an inhibitory effect on liver dysfunction, the content of egg white hydrolyzate in the food composition or the pharmaceutical composition is preferably from the results of Table 11. When the content is 0.01 to 30.0%, more preferably 0.05 to 10%, and still more preferably 0.3 to 5%, a superior in vivo antioxidant effect or liver function disorder suppressing effect can be obtained. I understand that.

Example 5 (In-vivo antioxidant food composition containing in-vivo antioxidant)
Cookies were prepared using the white powder in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1 with the following composition. Dried egg yolk (Kewpie Co., Ltd., dried egg yolk No. made by adding shortening and white sugar into a stirrer (Kitchen Aid, INC. Kitchen Aid K5SS), mixing with a speed control lever 6 for 1 minute to make a cream, and rehydrating with fresh water. 0.1) was gradually added, and stirring was further continued for 2 minutes. After mixing, the sieved flour, baking powder and in-vivo antioxidant were added, and stirring was continued for 1 minute to prepare a dough. The obtained dough was allowed to stand in a refrigerator for 2 hours, then extended to a thickness of about 3 to 5 mm, removed from the mold, and baked in an oven at 180 ° C. for 10 minutes to obtain cookies. In addition, the obtained cookie contains 1% of egg white hydrolyzate which is an active ingredient of the in-vivo antioxidant against the mass. I sampled these cookies and they were as good as regular cookies.

<Formulation (g)>
Flour 200
Baking powder 1
In vivo antioxidant 5
Shortening 120
Super white sugar 80
Dried egg yolk 16
Shimizu 20
――――――――――――――――
Total 442

Example 6 (In-vivo antioxidant food composition containing in-vivo antioxidant)
Using the white powdered in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1, cocoa with granular milk was prepared with the following composition. Each raw material powder was mixed in a V-type mixer and then granulated with an extrusion granulator to obtain cocoa containing granular milk. The content of egg white hydrolyzate which is an active ingredient of the in-vivo antioxidant of the obtained cocoa containing granular milk is 2% with respect to the product. The obtained cocoa containing granular milk is put into a 20 g cup, 150 mL of hot water at 98 ° C. is poured, and stirred with a spoon 30 times to prepare and taste a cocoa containing milk. As with commercially available milk cocoa, it was delicious.

<Formulation (g)>
Cocoa powder 210
Whole milk powder 240
Lactose 100
Sugar 430
In vivo antioxidant 20
―――――――――――――――
Total 1000

Example 7 (In vivo antioxidant pharmaceutical composition containing in vivo antioxidant)
300 g of white powder in vivo antioxidant material made of egg white hydrolyzate obtained in Example 1 and 100 g of crystalline cellulose (Asahi Kasei Co., Ltd., “Avicel”), dextrin (Matsuya Chemical Co., Ltd., “TK”) -16 ", DE value 16) 450 g and lactose 50 g were mixed with powder, and sucrose fatty acid ester (treated by Mitsubishi Chemical Foods Corporation," Ryoto Sugar Ester (S-570) ", HLB value 5) 4 g And 6 g of corn starch were added, and tableting was performed with a single-type tablet press. As a result, 820 g of a tablet-shaped pharmaceutical composition having a weight of 300 mg, a diameter of 9 mm, a layer thickness of 5 mm, and a hardness of 6.5 kp (more than 10 average values) was obtained. This contains 35% of egg white hydrolyzate which is an active ingredient of in vivo antioxidants.

Example 8 (Food composition for inhibiting liver dysfunction containing a liver dysfunction inhibitor)
A white powder in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1 was blended as a liver dysfunction inhibitor, and a cookie was obtained in the same manner as in Example 5. In addition, the obtained cookie contains 1% of an in vivo antioxidant material (egg white hydrolyzate), which is an active ingredient of a liver dysfunction inhibitor, with respect to mass. I sampled these cookies and they were as good as regular cookies.

Example 9 (Food composition for inhibiting liver dysfunction containing a liver dysfunction inhibitor)
A white powdered in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1 was blended as a liver dysfunction inhibitor, and cocoa with milk was obtained in the same manner as in Example 6. In addition, content of the in-vivo antioxidant material (egg white hydrolyzate) which is an active ingredient of the liver dysfunction inhibiting material of the obtained cocoa containing granular milk is 2% with respect to a product. When cocoa with milk was prepared and sampled in the same manner as in Example 6, it was as delicious as commercially available milk cocoa.

Example 10 (pharmaceutical composition for suppressing liver dysfunction including a liver dysfunction inhibitor)
A white powdered in-vivo antioxidant made of egg white hydrolyzate obtained in Example 1 was blended as a liver dysfunction inhibitor, and a tablet-like pharmaceutical composition was obtained in the same manner as in Example 7. This contains 35% of an in vivo antioxidant (egg white hydrolyzate) which is an active ingredient of a liver dysfunction inhibitor.

Claims (11)

  An in-vivo antioxidant comprising an egg white hydrolyzate as an active ingredient.   The in vivo antioxidant material according to claim 1, wherein the egg white hydrolyzate is treated with a neutral protease derived from Aspergillus.   The in vivo antioxidant material according to claim 2, wherein the egg white hydrolyzate is further treated with papain.   The in vivo antioxidant material according to any one of claims 1 to 3, wherein the in vivo antioxidant material is a lipid antioxidant material of a biological membrane.   The in vivo antioxidant material according to claim 4, wherein the lipid antioxidant material of the biological membrane is a HEL production inhibitor.   The in vivo antioxidant material according to claim 4 or 5, wherein the lipid antioxidant material of the biological membrane is a lipid antioxidant material of hepatocytes.   A food composition for in vivo antioxidant, comprising the in vivo antioxidant material according to any one of claims 1 to 6.   A pharmaceutical composition for in vivo antioxidants, comprising the in vivo antioxidant material according to any one of claims 1 to 6.   A liver dysfunction-suppressing material comprising the in vivo antioxidant material according to claim 6 as an active ingredient.   A food composition for inhibiting liver dysfunction, comprising the liver dysfunction inhibiting material according to claim 9. A pharmaceutical composition for inhibiting liver dysfunction comprising the liver dysfunction inhibiting material according to claim 9.