JP2020029401A - Liver function disorder inhibitor, food and drink product, pharmaceutical preparation, and method for producing liver function disorder inhibitor - Google Patents

Abstract

To provide a novel liver function disorder inhibitor, a food and drink product, and a pharmaceutical preparation which are derived from a natural product, and a method for producing a liver function disorder inhibitor.SOLUTION: A liver function disorder inhibitor contains a soybean fermented product by fungus containing at least one type of Rhizopus stolonifer and Rhizopus oryzae.SELECTED DRAWING: None

Description

  The present invention relates to a hepatic dysfunction inhibitor, a food and drink, a pharmaceutical, and a method for producing a hepatic dysfunction inhibitor.

  Liver disease with hepatic dysfunction develops due to viral infections, drugs, lifestyle-related diseases and the like. It is known that when these liver diseases become chronic and their symptoms progress, serious diseases such as liver cirrhosis and liver cancer eventually develop. For example, alcoholic or non-alcoholic fatty liver is known as a liver disease caused by lifestyle-related diseases. Fatty liver is a disease in which fat accumulates excessively in hepatocytes. In recent years, with the westernization of eating habits and the like, fat intake has increased, and patients with non-alcoholic fatty liver caused by a high-fat diet have increased. Non-alcoholic fatty liver may become non-alcoholic steatohepatitis when the disease progresses due to damage to hepatocellular tissue due to oxidative stress, etc., and may develop liver cirrhosis or liver cancer at the end stage It has been known. Under the circumstances described above, early suppression of liver dysfunction is important, and development of a highly effective liver dysfunction inhibitor for suppressing liver dysfunction is desired.

  Patent Literature 1 describes a liver dysfunction inhibitor containing, as an active ingredient, a water-soluble polymer substance obtained from a fermentation decomposition product obtained by fermenting a raw material containing soybean using koji mold. Patent Literature 2 discloses a liver function improving agent containing fermented steamed soybean and koji under pressure and heating, and containing salt-free miso as an active ingredient. In addition, in Non-Patent Document 1, when the tempe powder and the unfermented soybean powder were administered to two groups of white rats, the group to which the unfermented soybean powder was administered was compared with the group to which the tempe powder was administered. It is described that many pathological changes were observed.

Japanese Patent Application Laid-Open No. 2008-88151 (published on April 17, 2008) (Japanese Patent No. 4809785) JP-A-2015-15941 (released on January 15, 2015)

Japan Brewery Association Vol. 81, No. 8 (1986), 526-530

  In the prior art, Patent Literature 1, Patent Literature 2, and Non-Patent Literature 1 all describe that soybean-derived substances improve liver dysfunction. Among them, it is described that the hepatic dysfunction inhibitor of Patent Document 1 is particularly effective in suppressing iron-induced fatty liver and acute hepatic dysfunction caused by a drug of carbon tetrachloride. The main component is a high molecular substance obtained from soy sauce. Further, the liver function improving agent of Patent Document 2 needs to be fermented under pressurized heating, and requires equipment capable of pressurized heating. The tempeh used in Non-Patent Document 1 is a fermented soybean product obtained by fermenting soybeans with Rhizopus microsporus (hereinafter abbreviated as “R. microsporus”). In order to solve the above-mentioned health problems, there is a need to provide a novel hepatic dysfunction inhibitor that is more effective and easier to manufacture. In particular, it is useful to provide a novel hepatic dysfunction inhibitor derived from natural products.

  A main object of the present invention is to provide a novel liver function disorder inhibitor derived from a natural product, a food and drink, a pharmaceutical, and a method for producing a liver function disorder inhibitor.

  The present inventors have assiduously studied and, surprisingly, surprisingly found that Rhizopus stolonifer (hereinafter abbreviated as “R. stolonifer”) and Rhizopus oryzae (hereinafter “R. .Oryzae ")), a fermented soybean product obtained from a fungus containing at least one of the following was found to have an effect of suppressing hepatic dysfunction, thereby completing the present invention.

  That is, in order to solve the above problems, the present invention includes any one of the following embodiments.

  The agent for suppressing hepatic dysfunction according to one embodiment of the present invention includes a fermented soybean product of a fungus containing at least one of Rhizopus stolonifer and Rhizopus oryzae.

  A method for producing a hepatic dysfunction inhibitor according to another aspect of the present invention is a method for producing a hepatic dysfunction inhibitor, wherein the hepatic dysfunction inhibitor comprises at least one of Rhizopus stolonifer and Rhizopus oryzae. A method for producing a liver dysfunction inhibitor, comprising a step of producing a fermented soybean product by fermenting soybean using a fungus containing the fungus.

  ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of a novel liver function disorder inhibitor derived from a natural product, food and drink, a pharmaceutical, and a liver function disorder inhibitor can be provided.

  Hereinafter, embodiments of the present invention will be described in detail.

[1. Hepatic dysfunction inhibitor)
The agent for suppressing hepatic dysfunction according to one embodiment of the present invention includes R.I. stolonifer and R.S. soybean fermented product by fungi containing at least one of O. oryzae.

(Fermented soybeans)
The active ingredient of the liver dysfunction inhibitor according to the present invention is contained in a fermented soybean product. The fermented soybean product is described below. The fermented soybean product according to the present embodiment is R.R. stolonifer and R.S. It is a fermented soybean product by a fungus containing at least one of Oryzae. In other words, the fermented soybean product according to this embodiment is R. soybean. stolonifer and R.S. It may be obtained by fermenting soybeans using a fungus containing at least one of Oryzae.

  The material of the fermented soybean product according to the present embodiment, the method and conditions for production, and the like will be described in detail below.

(Fungi)
Fungi for fermenting soybean used in the present embodiment include fungi of the genus Rhizopus (Rhizopus).

  In this embodiment, soybean fermentation is used for R. soybean fermentation. stolonifer and R.S. oryzae which contains at least one of O. oryzae. Preferably, R. oryzae, R.A. stolonifer var. stolonifer (Rhizops stronifer) stolonifer var. lyococcus (Rhizopus stronifera liococcus). In addition, the place of origin of the fungus is not particularly limited. In addition to the above bacteria, microsporus var. oligosporus (Rhizopus microsporus). microsporus var. microsporus (Rhizopus microsporus); Rhizopus fungi such as delmar (Rhizopus derma), fungi included in commercially available tempeh, and bacteria of other genera such as Lactobacillus (lactobacillus) may be included. That is, a mixture of two or more of the above-mentioned bacteria may be used for soybean fermentation. Soybeans are fermented by adding fungi containing the above-mentioned bacteria to soybeans. The form of the bacterium to be added is not particularly limited as long as it is a vegetative mycelium or a spore, or a mixture thereof. Is preferred.

  Here, when the fungus is used in a combination of a plurality of different fungi, R.P./s. The proportion of the number of stronifer spores is preferably at least 30%, more preferably at least 40%, even more preferably at least 50%, particularly preferably at least 60%. Alternatively, R. per spore count of the spores of the fungus added to the soybean. The proportion of the number of spores of oryzae is preferably at least 30%, more preferably at least 40%, even more preferably at least 50%, particularly preferably at least 60%.

  R. stolonifer and R.S. or R. oryzae, for example, R. as the total number of spores of Rhizopus sp. stolonifer and R.S. As the ratio of oryzae, each bacterium may be contained in an equal amount.

  The soybean fermented product obtained by adding the above-described ratio of bacteria to soybean contains an active ingredient in an amount sufficient for use in the hepatic dysfunction inhibitor of the present invention.

(soy)
The form of the soybean used in the present embodiment may be a round soybean or a processed soybean, and a soybean hypocotyl or the like may be used.

(Soybean fermentation conditions)
In one embodiment, the fermentation of soybeans is carried out by inoculating the steamed soybeans with the above-mentioned fungi and performing solid culture. Hereinafter, each step and conditions in the fermentation will be described.

(Soybean pretreatment and steaming)
It is preferable to use soybeans immersed in a liquid before steaming. Preferred liquids for immersion include water or edible organic acids, and aqueous solutions (such as vinegar and brewed vinegar) containing foods containing these organic acids. Among them, the edible organic acid is preferably at least one organic acid selected from acetic acid, citric acid, malic acid, succinic acid and fumaric acid. The concentration of the organic acid in the aqueous organic acid solution is not particularly limited, but is preferably such that the growth of bacteria added to soybean is not inhibited, for example, 0.1 to 0.5% (w / w). The degree is preferred. In addition, soybeans are preferably shed before, after immersion or after steaming in a liquid, and more preferably soybean outer skin does not remain during inoculation. The temperature and time of the steaming are not particularly limited, but, for example, it is preferable that the soybeans are immersed in water and boiled in water for about 5 to 90 minutes or pressure cooked at 105 to 120 ° C for about 2 to 30 minutes. After steaming, the soybeans are cooled and the bacteria are inoculated.

<Bacterial inoculation and culture>
For inoculating the steamed soybeans, a spore suspension, a spore powder or the like is preferably used, and preferably 1 × 10 ³ to 1 × 10 ⁹ , more preferably 1 ×, per 100 g (wet weight) of the steamed soybeans. Add 10 ⁴ to 1 × 10 ⁸ spores.

  Here, when a plurality of bacterial species are used, the spores may be mixed in advance, or may be separately added to the steamed soybeans for each bacterial type.

<Culture conditions>
After the inoculation, the cells are mixed and filled in, for example, a polybag having one or more holes, or left in a stainless steel tray, and solid-cultured by a usual method. Preferably, the cells are cultured aerobically in the presence of oxygen.

  As the culture conditions, the culture temperature is preferably 20 to 42 ° C, more preferably 25 to 37 ° C, and still more preferably 28 to 34 ° C. The cultivation time varies depending on the temperature, the amount of inoculation, and the like, but is preferably 10 to 60 hours, more preferably 15 to 50 hours, and still more preferably 20 to 40 hours. The culture is performed under normal pressure (atmospheric pressure). The culturing can be performed under a pressure of, for example, 0.1 MPa to 100 MPa.

  By culturing the bacterium under the above conditions, the hepatic dysfunction inhibitory component in the fermented soybean product can be increased.

  The fermented soybean product of the present invention can be obtained by performing fermentation under the above conditions. Further, the fermented soybean product obtained by fermenting soybean under the above-mentioned conditions contains a high concentration of an active ingredient having a hepatic dysfunction inhibitory activity in the hepatic dysfunction inhibitor of the present invention.

  The fermented soybean product obtained can be stored frozen. In addition, drying may be performed, and drying may be performed by air drying, spray drying, vacuum drying, freeze drying, or a combination of these methods. Furthermore, it may be sterilized or sterilized before or after drying. The obtained fermented soybean may be sterilized or sterilized by a method such as wet heat heating, dry heat heating, electromagnetic wave treatment, or high pressure treatment, for example, heat treatment at 60 to 121 ° C. for 1 minute to 6 hours. By sterilization or sterilization, propagation of various bacteria can be suppressed and safety and health can be ensured. Further, decolorization and / or deodorization treatment may be performed by any method. By drying, it becomes easy to pulverize the dried product by pulverizing it. The composition obtained by performing the decolorizing step, the deodorizing step, and the like has a high versatility and can be suitably used for various uses such as food and drink and pharmaceuticals described below.

(Other components of liver dysfunction inhibitor)
Soybeans or fermented soybeans are further rich in proteins, dietary fiber, amino acids, vitamins (for example, vitamin E and vitamin B group), minerals such as iron, lecithin, saponin, isoflavones, etc. It is rich in nutritional value and contains various functional ingredients. Therefore, in one embodiment, the hepatic dysfunction inhibitor has at least one of various functional components derived from soybean or fermented soybeans, as well as a component having hepatic dysfunction suppressing activity. Therefore, the hepatic dysfunction inhibitor may have not only a hepatic dysfunction-suppressing effect but also a health-promoting effect of one or more known or novel components contained in soybeans or soybean fermented products.

(Liver dysfunction)
As used herein, “liver dysfunction” refers to a state in which liver function is abnormal. The state in which liver function is abnormal refers to a state in which the function is lower than that of a normal liver. In one embodiment, the decrease in liver function is determined by a numerical value of an index of liver function disorder described below. As used herein, the term “hepatic dysfunction” includes a state in which liver disease has not been developed but liver function is abnormal, and a liver disease exhibiting liver dysfunction. Hepatic dysfunction includes alcohol-induced hepatic dysfunction, non-alcohol-induced hepatic dysfunction, drug (halogen compounds, galactosamine, etc.)-Induced hepatic dysfunction, and the like.

  In one embodiment, the liver dysfunction inhibitor can be used effectively for the prevention or treatment of the various liver dysfunctions described above. As used herein, "prevention" refers to preventing a person from suffering from a disease, disease or disorder. “Treatment” refers to alleviation or elimination of a disease, disorder or disorder that is already suffering, or symptoms associated therewith.

  The hepatic dysfunction inhibitor of the present invention is particularly preferably used for nonalcohol-induced hepatic dysfunction, and more preferably for hepatic dysfunction induced by a high fat diet. Here, a high-fat diet means a diet that is rich in fat. For example, in the “Standards of Daily Dietary Intake of Japanese (2015 version)” set by the Ministry of Health, Labor and Welfare, the standard of daily lipid intake is , About 80 g / day for men and about 70 g / day for women, which means that the diet is a dietary intake of lipids above this intake standard.

  In one embodiment, the liver dysfunction to which the liver dysfunction inhibitor is applied is preferably non-alcoholic fatty liver, non-alcoholic liver dysfunction such as non-alcoholic steatohepatitis, and more preferably non-alcoholic fatty liver. The liver. Also, these non-alcoholic liver dysfunctions may be induced by a high fat diet.

  In addition, the hepatic dysfunction inhibitor has not only the function of suppressing the dysfunction of the liver itself, but also the effect of preventing or treating complications accompanying the hepatic dysfunction. Complications include dehydration, hepatic encephalopathy, hepatic diabetes, esophageal varices, gastric ulcer and the like. Furthermore, applying the liver dysfunction inhibitor to prevent the above-mentioned hepatic dysfunction from occurring is to suppress hepatic dysfunction at an early stage, to prevent cirrhosis and prevent aggravation of symptoms to liver cancer and the like. It has clinical significance.

<Evaluation of the effect of suppressing liver dysfunction>
The hepatic dysfunction inhibitory effect of the hepatic dysfunction inhibitor of the present invention can be evaluated by various hepatic dysfunction indexes generally used in liver function tests.

  As an index of liver dysfunction, first, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are mentioned. Another indicator of liver dysfunction is lactate dehydrogenase (LDH). Further, typical indices representing other liver function states include total bilirubin and ammonia.

  Hepatic dysfunction can be determined based on whether or not the measured value of any one or more of the above-mentioned indices deviates from a normal value (reference value) range. The determination is made based on, for example, the presence or absence of a significant difference from the reference value. As the reference value, a predetermined reference value determined in advance by a laboratory or the like may be used, and for example, a value obtained from a normal human or animal having no liver dysfunction may be used.

(Form of liver dysfunction inhibitor)
In one embodiment, the hepatic dysfunction inhibitor has a soybean fermented product as an active ingredient, as described above, and the hepatic dysfunction inhibitor in one embodiment contains the soybean fermented product itself or all components in the soybean fermented product. The dried product of the fermented soybean product or the product containing all the components in the dried product of the fermented soybean product may be used. In addition, it can be formulated into a suitable form and used according to each use such as a medicine, food and drink. Furthermore, it can be blended with known foods and drinks and pharmaceuticals. Details of additives and the like in the case of use for pharmaceuticals and foods and beverages are described in the following items. Further, the administration method, administration form, dosage amount and the like of the hepatic dysfunction inhibitor to the subject will be described in detail in the following items.

[2. Drugs containing hepatic dysfunction inhibitors)
The medicine according to one embodiment of the present invention contains a hepatic dysfunction inhibitor. Hepatic dysfunction inhibitors contained in pharmaceuticals include [1. Liver dysfunction inhibitor] as described above.

  Since the hepatic dysfunction inhibitor of the present embodiment has a remarkable hepatic dysfunction inhibitory activity, it is possible to prevent or treat hepatic dysfunction by administering a drug containing the hepatic dysfunction to a subject. Therefore, the use of the pharmaceutical product of the present embodiment is useful, for example, for the treatment of a patient having a disease, disorder or disorder exhibiting liver dysfunction in a living body. For specific examples of liver dysfunction, see [1. Hepatic dysfunction inhibitor]. In addition, the medicament of the present invention can be used for therapeutically or prophylactically treating subjects such as animals and humans at an increased risk of developing liver dysfunction.

  In one embodiment, the pharmaceutical composition of the present invention further includes at least one substance selected from a suitable solvent, carrier, excipient, adjuvant, and the like. Hereinafter, each component constituting the pharmaceutical is described. In addition, as long as the effect of the active ingredient of the drug according to the present invention is not inhibited, it may contain a known other hepatic dysfunction inhibitor or an active ingredient having another effect.

(solvent)
Examples of the solvent of the drug of the present invention include water, buffer, and the like, and examples of the buffer include physiological saline, phosphate buffer, Ringer's solution, and the like. Further, the solvent used in the composition may be a mixture of two or more of the above solvents.

(Carrier)
The carrier used for the pharmaceutical product of the present invention is not particularly limited, and various kinds of desired carrier substances known as materials for pharmaceutical preparations can be used. The carrier substance is not particularly limited. For example, in a solid preparation, an excipient, a lubricant, a binder, a disintegrant and the like can be mentioned. In addition to the above carrier substances, pharmaceutical additives such as preservatives, coloring agents, natural pigments, sweeteners and the like can also be used if necessary.

(Formulation)
The dosage form of the medicament of the present invention is not particularly limited, and may be liquid, solid, semi-solid or semi-liquid. These dosage forms can be easily produced based on a method known to those skilled in the art. When the dosage form is a liquid, for example, preparations, syrups, and the like in which the hepatic dysfunction inhibitor of the present invention is dispersed, suspended or dissolved in a substantially aqueous solvent are included. Here, a substantially aqueous preparation or preparation refers to, in one embodiment, a composition containing a certain ratio of one or more non-aqueous components or one or more other pharmaceutically acceptable components. Intended. When the dosage form is solid, powder, granules, tablets, capsules (hard capsules, soft capsules, microcapsules) and the like can be mentioned. When the dosage form is a semi-solid or semi-liquid, examples thereof include ointments, lotions, creams, gels, suspensions, and emulsions.

  The above-mentioned pharmaceuticals can be manufactured by using a conventional method in the technical field of formulation technology.

  In another embodiment, the medicament can also be produced by adding the hepatic dysfunction inhibitor of the present invention to various known medicaments.

(Administration route)
Routes of administration of pharmaceuticals include, but are not limited to, oral, topical, subcutaneous, intramuscular, intravenous, intradermal, transdermal, and the like. Among them, oral administration is preferred from the viewpoint of easy administration.

(Method of administration)
Administration of the pharmaceutical agent of the present invention to a subject can be performed by subcutaneous, intravenous, intramuscular, direct injection into the intraperitoneal cavity, intranasal, intraoral, intrapulmonary, intravaginal, rectal, etc. It is performed based on known methods such as oral administration and intravascular administration.

(Dose and administration period)
The dose (therapeutically effective amount) of the drug can be appropriately selected depending on the age, body weight, and sex of the living body to be administered, the dosage form to be administered, the desired degree of the effect of suppressing hepatic dysfunction, and the like. Also, the administration period of the drug can be appropriately selected so as to obtain the intended effect of suppressing liver dysfunction. The administration period is, for example, preferably 1 day to 90 days, more preferably 3 days to 30 days. The number of times of administration is preferably 1 to 3 times per day, more preferably 1 to 2 times. During the administration period, it may be administered continuously every day or once every 2 to 4 days.

  In addition, for example, as long as the hepatic dysfunction-suppressing effect of the hepatic dysfunction-suppressing agent is not impaired, it may be administered in combination with another drug having a hepatic dysfunction-suppressing effect or a drug having another effect.

(Use applications)
A specific use of the medicament of the present invention is, for example, the treatment or prevention of the above-mentioned diseases, diseases or disorders exhibiting liver dysfunction. [1. Liver dysfunction inhibitor] as described above.

(Target of administration)
The subject to which the medicament according to the present invention is administered includes all animals, preferably vertebrates such as mammals and birds, and more preferably mammals. Further, it is preferred that the mammalian subject is a human, but livestock, laboratory animals or pet animals may be subjects. Specific examples include domestic animals such as chickens, pigs, horses, goats, sheep and cattle, pet animals such as cats, dogs, hamsters, rabbits and guinea pigs, mice, monkeys, fish, birds and the like.

  Since the active ingredient for suppressing liver dysfunction is naturally derived and has high safety, the medicament of the present invention is suitably used for long-term administration. Therefore, for example, for the prevention or treatment of fatty liver caused by a high-fat diet or a lifestyle-related disease, it is possible to take it for a long time.

[3. Treatment method for liver dysfunction)
(Administration method / dose)
The present invention also includes a method for treating hepatic dysfunction, which comprises the step of administering a therapeutically effective amount of a drug containing the above-mentioned hepatic dysfunction inhibitor to a subject. In one embodiment, the subject to be administered has impaired liver function. Here, as the hepatic dysfunction inhibitor, the hepatic dysfunction inhibitor alone may be administered alone, or may be administered as one component of a pharmaceutical composition suitable for the purpose of administration. The administration method, dosage, administration period, etc. are described in [2. Drugs Containing Hepatic Dysfunction Inhibitor]. In addition, the form of prophylactic administration, which includes the step of administering the drug to a healthy subject before the onset of liver dysfunction, is also included in the category of the therapeutic method of the present invention. That is, a mode in which the hepatic dysfunction inhibitor is administered to a subject in an amount equal to or more than a therapeutically effective amount in a living body to prevent hepatic dysfunction in advance.

(Combination therapy)
In the method for treating hepatic dysfunction according to the present invention, a combination therapy may be performed by administering in combination with a hepatic dysfunction inhibitor other than the hepatic dysfunction inhibitor of the present invention (for example, known hepatic dysfunction suppression). By performing the combination therapy, a therapeutic effect synergistic with the therapeutic effect of the combination drug can be expected.

[4. Food and drink containing hepatic dysfunction inhibitor)
A food or drink according to one embodiment of the present invention contains the hepatic dysfunction inhibitor according to the present invention. As used herein, the term "food and drink" includes not only beverages, foods and health foods consumed by humans, but also feeds or feeds fed to animals such as domestic animals and pets. Further, the food or beverage may further contain additives other than the liver dysfunction inhibitor. Hepatic dysfunction inhibitors contained in foods and drinks include [1. Liver dysfunction inhibitor] as described above. The amount of the hepatic dysfunction inhibitor contained in the food and drink depends on the type and form of the food and drink, and is not particularly limited.

  As other components contained in the food or drink, any desired additive may be used as long as it does not impair the hepatic dysfunction inhibitory effect of the hepatic dysfunction inhibitor, for example, a pH adjuster, an organic acid, a sugar alcohol, Sweeteners, flavors, vitamins, bone metabolism vitamins, antioxidants, excipients, solubilizers, binders, lubricants, suspending agents, wetting agents, film-forming substances, flavoring agents, flavoring agents, coloring Additives commonly used in compositions such as additives, preservatives, antibacterial agents, bactericides, anti-inflammatory agents, and the like.

(Form of food and drink)
The form of the food and drink composition is not particularly limited, and may be, for example, a pharmaceutical form, processed food, and various beverages. Examples of the formulation include health foods, functional foods, foods for specified health use, and the like. In addition, the food for specified health use (including the food for specified health use with conditions) is a food that can be displayed on its packaging container or the like with a function indication or a health use indication approved or approved by the Ministry of Health, Labor and Welfare of Japan. When the "food or drink" of the present invention is in the form of a preparation, it can be configured in accordance with a pharmaceutical. That is, the content of the hepatic dysfunction inhibitor in the food or drink can be adjusted to the content according to the drug, and the dosage form can be adjusted to the drug, such as tablets, capsules, drinks, and powders.

  The food or drink according to the present invention suppresses hepatic dysfunction in a living body by orally ingesting the body. Therefore, it is preferable that the food or drink is in a form that can be easily taken orally. For example, known drinks such as tea, coffee, black tea, soy milk, powdered juice, confectionery such as chewing gum, biscuit, candy and gummy candy, or troches, tablets, capsules, tablets, and formulated forms such as granules can be mentioned. Can be

  In one embodiment, foods and drinks can be manufactured by adding to the above-mentioned various known foods and drinks.

[5. Method for producing hepatic dysfunction inhibitor)
The present invention further relates to a method for producing a hepatic dysfunction inhibitor, the hepatic dysfunction inhibitor comprising R.R. stolonifer and R.S. The present invention provides a method for producing a hepatic dysfunction inhibitor, which comprises a step of producing a fermented soybean product by fermenting soybean using a fungus containing at least one of Oryzae.

  In one embodiment, R.I. stolonifer and R.S. a step of producing a fermented soybean product by fermenting a soybean using a fungus containing at least one of S. oryzae (a soybean fermented product production process) and mixing the fermented soybean product with other components to thereby enhance the liver. A step of preparing a dysfunction inhibitor (hepatic dysfunction inhibitor preparation step) is included.

  In still another embodiment, the method for producing a hepatic dysfunction inhibitor may further include a step (drying step) of drying the fermented soybean product obtained in the above-mentioned soybean fermentation step. As another further step in the production method, at least one of the heating step and the sterilizing step may be performed after the above-described soybean fermented product producing step.

  ADVANTAGE OF THE INVENTION According to the manufacturing method of the hepatic dysfunction inhibitor which concerns on this invention, the hepatic dysfunction inhibitor which can be used for various wide-ranging uses including the active ingredient with a high hepatic dysfunction inhibitory effect can be manufactured.

  The present invention is not limited to the embodiments described above, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention.

  Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited to the following examples.

[Example 1: Species R. sp. Preparation of fermented soybean powder of stolonifer]
To 100 g of the dehulled soybean, a 5% aqueous brewed vinegar solution of 5 times the weight was added, and immersion treatment was performed at room temperature overnight. After the immersion treatment, 160 g of immersed soybeans were obtained by draining water. 320 g of tap water was added to 160 g of immersed soybeans, and the mixture was steamed at 100 ° C. for 10 minutes using a pressure cooker. After the steaming treatment, 200 g of steamed soybeans were obtained by draining the water. For 100 g of steamed soybean cooled to 40 ° C. or less, R.I. A spore suspension of stolonifer NBRC 30816 was added so that the number of spores became 1 × 10 ⁶ spores, and the mixture was thoroughly mixed to perform seeding treatment. This was filled in a perforated bag and subjected to a fermentation treatment for 40 hours in a thermostat set at 32 ° C. After the fermentation treatment, sterilization was performed at 90 ° C. for 30 minutes, and further freeze-dried to obtain a fermented soybean powder (Example 1).

[Example 2: Bacterial species R. Preparation of fermented soybean powder of oryzae]
Instead of the spore suspension in Example 1, R. A fermented soybean powder was obtained in the same manner as in Example 1 except that a spore suspension of oryzae NBRC4716 was used (Example 2).

[Comparative Example 1: Bacterial species R. Preparation of powder of fermented soybean of microsporus]
Instead of the spore suspension in Example 1, R. A fermented soybean powder (tempe powder) was obtained in the same manner as in Example 1 except for using a spore suspension of microsporus NBRC32002 (Comparative product 1).

[Example 1: Evaluation test for suppression effect of high-fat diet-induced hepatic dysfunction by administration of soybean fermented product powder]
(Test method)
Four-week-old Sprague Dawley male rats with an average body weight of 109 g were divided into 4 groups of 10 rats each, and after 1 week of preliminary breeding (fed with a normal commercial feed), Examples 1 and 2 and Comparative product 1 A high-fat diet (30% by weight of fat) containing 20% by weight of the fermented soybean powder per feed weight was continuously ingested daily for 21 days. In addition, in place of the soybean fermented product powder, casein was added as a control so that the protein content (20% by weight) became equal (Comparative Product 2). After the rearing, AST, ALT, total bilirubin, ammonia, and LDH in serum, which were test items for liver dysfunction, were measured. Table 1 shows the measurement results. The rate of increase / decrease in the measured values of the comparative examples 1 and 2 and the measured values of the comparative product 1 is shown in parentheses.

(Test results)
R. Example 1 which is a soybean fermented product powder obtained using stolonifer was statistically significantly impaired in AST, total bilirubin and ammonia in comparison with Comparative Example 2 which was a control casein intake group. Was suppressed. In this test, Tukey's range test was used to test for a significant difference for any index. Further, in the case of ALT and LDH, the value of Example 1 was lower than that of Comparative Example 2, indicating a tendency to suppress liver dysfunction.

  In addition, R. Example 2, a soy fermented powder obtained using Oryzae, statistically significantly improved liver dysfunction in ammonia and control casein intake in AST, ALT, total bilirubin and LDH. The value was lower than that of the comparative product 2, which was a group, and was in an improving tendency.

  On the other hand, R. Comparative product 1 which is a tempeh powder obtained using microsporus is improved in other test items for liver dysfunction except that it has a low value in ammonia as compared with comparative product 2 which is a control casein intake group. Was not observed. Table 1 summarizes the above results. In the table,% in parentheses indicates the rate of increase / decrease with respect to Comparative Product 2.

[Reference Example: Test for evaluating the content of ethanol-insoluble substances in soybean fermented product extracts of the actual product and the comparative product]
(Test method)
1. Preparation of Extract Solution To a 50 mL centrifuge tube, 2 g of each of the soybean fermented product powders of Example 1, Example 2 and Comparative Example 1 and 20 mL of 25% ethanol were added, and the mixture was subjected to shaking extraction at room temperature for 2 hours. (3,500 rpm, 10 minutes), and the supernatant was passed through a 0.45 μm filter to obtain an extract of each fermented soybean product.
2. Fractionation by Ultrafiltration 10 mL of the prepared extract was subjected to fractionation by centrifugation using an ultrafiltration membrane having a molecular weight cutoff of 5,000. The non-permeated liquid was collected in a 10-mL volumetric flask, and the volume was adjusted using 25% ethanol to obtain a fraction of each soybean fermented product extract (molecular weight: 5,000 or more).
3. Removal of ethanol-soluble substance Transfer all 10 mL of the soybean fermented product extract fraction (molecular weight: 5,000 or more) to a 50-mL centrifuge tube whose weight was previously weighed, add 20 mL of ethanol thereto, and vortex for 30 seconds. After standing at room temperature for 30 minutes, solid-liquid separation was performed by centrifugation (3,500 rpm, 10 minutes). After removing the supernatant, the remaining ethanol in the centrifuge tube was removed by heating in a constant temperature bath at 80 ° C., followed by freeze-drying, and measuring the weight of the dried centrifuge tube to obtain a molecular weight of 5,000. The weight of the above ethanol-insoluble substance was calculated.

(result)
R. stolonifer or R.S. oryzae containing ethanol-insoluble substances having molecular weights of 5,000 or more (Examples 1 and 2) contained in the soybean fermented product powder obtained using R. oryzae. The content was lower than that of the ethanol-insoluble substance having a molecular weight of 5,000 or more (Comparative Product 1) contained in the powder of the fermented soybean by microsporus. On the other hand, according to the results of the examples, the hepatic dysfunction-suppressing effect was determined by Example 1 which is a soybean fermented product powder obtained using stolonifer has the best effect, oryzae, an effect was observed in Example 2, which was a soybean fermented product powder obtained using R. oryzae. No effect was observed in Comparative Product 1, which is a fermented soybean powder obtained using microsporus.

  From the above, there is no correlation between the weight of an ethanol-insoluble substance having a molecular weight of 5,000 or more and the effect of suppressing hepatic dysfunction, and the active ingredient for inhibiting hepatic dysfunction in the fermented soybean product according to the present invention is described in Patent Document 1. It was presumed that it was not an ethanol-insoluble polymer substance.

  The hepatic dysfunction inhibitor according to the present invention has a hepatic dysfunction inhibitory effect, and can be used in a wide range of fields such as testing and research as well as food and drink and pharmaceutical applications.

Claims (7)

  An agent for inhibiting hepatic dysfunction, comprising a fermented soybean product from a fungus containing at least one of Rhizopus stolonifer and Rhizopus oryzae (Rhizopus oryzae).   The liver dysfunction inhibitor according to claim 1, wherein the fermented soybean product is fermented under normal pressure.   The hepatic dysfunction inhibitor according to claim 1 or 2, wherein the hepatic dysfunction is a non-alcoholic hepatic dysfunction.   The hepatic dysfunction inhibitor according to any one of claims 1 to 3, wherein the non-alcoholic hepatic dysfunction is caused by a high-fat diet.   A food or drink comprising the hepatic dysfunction inhibitor according to any one of claims 1 to 4.   A pharmaceutical comprising the hepatic dysfunction inhibitor according to any one of claims 1 to 4. A method for producing a liver dysfunction inhibitor,
A method for producing a hepatic dysfunction inhibitor, comprising: producing a fermented soybean product by fermenting soybean using a fungus containing at least one of Rhizopus stolonifer and Rhizopus oryzae. .