JP5993112B2 - Antidepressant - Google Patents

Description

  The present invention relates to an antidepressant used for the improvement or prevention of mental depressive symptoms such as depression, anxiety, fear, reduced motivation, and irritation.

  In the modern stress society, the number of patients with depression is increasing, and prevention and treatment measures are desired. Until now, the main mechanism of action of antidepressants was inhibition of monoamine reuptake. However, there are reports that conventional antidepressants are not effective in about 30% of patients, and antidepressants with a new mechanism of action are required. In addition, an antidepressant that can be safely taken without side effects is desired.

  On the other hand, it is conventionally known to use edible mushrooms as a component of an antidepressant. For example, Patent Document 1 contains a powder or extract of at least one food material selected from the group consisting of spices mainly composed of hop extract, licorice, toki, seri-root, and mushrooms, and is resistant to depression. The invention of an antidepressant food characterized by having a depressing action is disclosed, and mushrooms include dried shiitake mushrooms, fresh product shimeji, etc. (Claims of Patent Document 1, (See paragraph 0012). Patent Document 2 discloses an invention of a preventive or ameliorating agent for psychiatric symptoms of mild depression or depression, characterized by blending maitake (see claim 4 of Patent Document 2). .

JP 2002-58450 A JP 2005-325096 A

  However, an antidepressant comprising a mushroom fruit body powder or water extract as an active ingredient has not always been sufficiently effective.

  Therefore, an object of the present invention is to provide an antidepressant that has a basidiomycete-derived component such as mushrooms as an active ingredient and can be safely taken without side effects, and has a high effect of improving or preventing depressive symptoms.

  In order to achieve the above object, the present invention cultivates mycelia of basidiomycetes using a medium containing plant fiber raw materials, and extracts carbohydrates, proteins, and water-soluble lignin obtained by extraction from the culture. The present invention provides an antidepressant characterized by containing an extract containing it as an active ingredient.

  In the antidepressant of the present invention, it is preferable that the extract is obtained by performing autolysis of basidiomycetes from the culture.

  In addition, the plant fiber raw material is preferably prepared from a plant family plant, one selected from bagasse, corn stover, wheat bran, rice bran, rice bran, rice bran, bear cocoon, and bamboo or More preferably, it is prepared from two or more.

  Furthermore, it is preferable that the basidiomycete is selected from mannenpox, bukkyou, kofukisarokoshikake, kawara mushroom, shiitake mushroom, hira mushroom, mai mushroom, enoki mushroom, shimeji mushroom, yamabushi mushroom, and agaricus.

  When a mycelium of basidiomycetes is cultured using a medium containing plant fiber raw materials, cellulose, hemicellulose, lignin, etc. contained in the medium are cellulase, phenol oxidase, and laccase produced by mycelia of basidiomycetes during the culture period. Pentose-based proteoglycans are produced by digestion, degradation, and condensation with enzymes such as peroxidase and protease. Moreover, the water-soluble modified water-soluble lignin is generated. According to the present invention, mental extracts such as depression, anxiety, fear, reduced motivation, and irritability are obtained by using an extract containing the components from the culture medium in this way as an active ingredient. It is possible to provide an excellent antidepressant that can be expected to have an effect of improving or preventing the above. Moreover, the active ingredient is an extract obtained from a natural product, and can be safely taken on a daily basis without side effects.

It is a graph which shows the result of the basic examination of the elevated plus maze test by diazepam administration. It is a graph which shows the result of an elevated plus maze test when MAK or imipramine is administered. It is a graph which shows the result of the basic examination of the forced swimming test by imipramine administration. It is a graph which shows the result of a forced swimming test when MAK or imipramine is administered.

  The basidiomycetes used in the present invention are not particularly limited, for example, medicinal mushrooms such as mannen mushrooms, bukkyou, kofukisarokoshikake, kawara mushrooms, shiitake mushrooms, Japanese cypress mushrooms, mai mushrooms, enoki mushrooms, shimeji mushrooms, yamabushi mushrooms, Various foods such as edible rice cakes such as Agaricus are listed. Among these, mannen mushrooms and shiitake mushrooms are particularly preferably employed.

  In the present invention, mycelia of these basidiomycetes are cultured using a medium containing plant fiber raw material, and active ingredients are extracted from the culture. In this case, either a solid medium or a liquid medium can be used as the medium. As the plant fiber raw material used for the medium, those prepared from plants containing lignin are preferably used. As the lignin-containing plant, a plant family such as bagasse (squeezed sugar cane), corn stover, wheat bran, rice bran, rice straw, and straw are preferably used. In addition, bears, bamboo, etc. can be used. Particularly preferably, a culture medium containing at least one selected from bagasse, bear stems and corn stalks and rice bran is used. In addition, yeast extract, dry yeast, chlorella, spirulina, corn meal, okara, and the like may be added and mixed with the medium as other nutritional components as necessary.

  The mycelium of basidiomycetes is cultured by inoculating the mycelium of basidiomycetes in a medium containing the above-mentioned plant fiber raw material. In the case of a solid medium, the water content is adjusted to 60 to 80%, sterilized by autoclaving according to a conventional method, inoculated with mycelia, and, for example, 3 to 6 in a culture room conditioned at a temperature of 18 to 25 ° C. Incubate for months. The medium in which the mycelium has spread is preferably transferred to a temperature treatment chamber and subjected to a temperature change treatment. In the temperature change treatment, for example, the mixture is first heated at 30 to 34 ° C. for 24 to 48 hours, and then transferred to a low temperature chamber and treated for 3 to 5 days. After that, when it is moved to a culture room, the generation of fruiting bodies starts. At this point, the culture is terminated and the culture is crushed with a crusher.

  On the other hand, in the case of a liquid medium, after pulverizing the plant raw material as described above, adding other nutritional components such as rice bran as necessary, and preparing the medium so that the raw material is 5 to 20% by mass, The culture is preferably carried out by aeration and agitation culture or shaking culture at a temperature of 20 to 28 ° C. for about 1 week to 2 months. The culture is terminated when the pH of the medium is lowered to 3.5 to 5 and the mycelium is prevalent in the medium.

  After completion of the culture, the culture is extracted. As a preferable method, it is preferable to self-digest the mycelium and extract the culture using an enzyme present in the mycelium.

Specifically, in the case of a solid medium, the culture after culturing is crushed, a small amount of water is added as necessary, and the mixture is treated at 30 to 60 ° C. for 3 to 6 hours to advance the mycelial enzyme reaction, Let yourself digest. Subsequently, this crushed material is infiltrated with warm water or hot water of 50 ° C. or higher, and an active ingredient is extracted. The extraction can also be carried out under a high pressure such as 120 ° C. under a vapor pressure of 1.2 kg / cm ² , for example. The extract suspension thus obtained is preferably filtered or centrifuged, and the filtrate or supernatant is collected to obtain an extract containing a degradation product of the culture medium, a mycelium metabolite, a mycelium cell degradation product, and the like. Can be obtained.

On the other hand, in the case of liquid culture, the whole culture is treated at 30 to 60 ° C. for 3 to 6 hours to self-digest the mycelium to obtain a liquid suspension culture. Then, if necessary, a small amount of water is added, and the mixture is heated to 50 ° C. or higher, possibly under high pressure (for example, under a vapor pressure of 1.2 kg / cm ² ), and the extract is collected. By filtering or centrifuging the extract as necessary, and collecting the filtrate or supernatant, an extract containing a degradation product of a medium, a mycelium metabolite, a mycelium cell degradation product, or the like is obtained. Can do.

The extract thus obtained can be used as it is or after being concentrated, but can also be used by pulverizing it by means such as freeze drying or spray drying. When using those liquid as such or concentrated to as extract, Ru can also be, for example, a liquid or jelly drink type products. When utilizing the extract was triturated a conventional method by, powders, granules, tablets, Ru can be manufactured as capsules and the like.

  The above extract, which is an active ingredient of the present invention (a component extracted from a culture obtained by culturing mycelia of basidiomycetes in a medium containing plant fiber raw material) is a substance mainly composed of carbohydrates. However, it is preferable to have the following physicochemical properties.

(1) Molecular weight: 1 million or less (2) Chemical composition:
・ Sugar: 30-50% by mass
・ Protein: 8-20% by mass
Water-soluble lignin: 20-40% by mass

  In addition, in order to specify the range of the composition of the saccharide, protein, and water-soluble lignin in the above extract, it is specified by the mass composition obtained by measurement by the phenol sulfuric acid method, the semi-micro Kjeldahl method, and the acetyl bromide method, respectively. It is preferable.

  The results of testing the safety of the above culture extract (powder) obtained using mannen koji are as follows.

(A) Acute toxicity test (minimum lethal dose)
・ Rat single oral administration Male: 22,500mg / Kg or more
Female: 22,500 mg / Kg or more ・ Single mouse oral administration Male: 2,000 mg / Kg or more
Female: 2,000 mg / Kg or more (B) Rat 3-month repeated oral administration test (maximum no-effect level)
Male: 3,610 mg / Kg
Female: 4,190mg / Kg

  The effective dose of the antidepressant of the present invention is 1 to 10 g per day for an adult in terms of oral intake in terms of solid content of the extract. If the dose is smaller than this, the effect of the antidepressant action is not sufficiently obtained, and if the dose is larger than this, loose stool or abdominal bloating may occur. However, there is no problem in safety even if the dose is larger than the above.

  EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples, but these examples do not limit the scope of the present invention.

(1) Preparation Example of Culture Medium Extract <Preparation Example 1> (Solid culture of N. gonorrhoeae using bagasse)
90 parts by mass of bagasse and 10 parts by mass of defatted rice bran were mixed, adjusted to a moisture content of 70%, a solid medium was prepared, and autoclaved as usual. This solid medium is inoculated with mycelium of mannen moth and cultured in a culture chamber adjusted to 25 ° C. for 3 months. After the mycelium spreads in the medium, it is transferred to a temperature treatment chamber and heated at 35 ° C. for 24 hours. Then, it was treated in a low temperature room at 10 ° C. for 3 days. Then, it culture | cultivated for 3 days in the said culture room, and crushed the culture medium in the magnitude | size about the thumb with the crusher. The crushed medium was treated at 40 ° C. for 6 hours to promote autolysis, then packed in an extraction tank, and extracted for 16 hours while circulating 60 ° C. warm water. The obtained extract was filtered through a cartridge filter, further sterilized by filtration through a membrane filter, concentrated, and freeze-dried to obtain a brown powder. The results of analyzing the components were as follows.

・ Sugar: 40% by mass (phenol sulfate method)
・ Protein: 12% by mass (semi-micro Kjeldahl method)
・ Water-soluble lignin: 30% by mass (acetyl bromide method)
・ Inorganic: 13% by mass (direct ashing method)

(2) Animal test The animal test using the rat (Sprague Dawley, Sankyo Lab Service) for the mycelium culture medium extract (hereinafter referred to as “MAK”) of mannen koji obtained by the method of Preparation Example 1 above. Thus, the antidepressant effect was examined. Specifically, MAK was administered to rats in an oral sonde once a day at a dose of 3 mL / kg at a dose of 1 g / kg, and this was continued for 15 days to form test groups (6 animals). Moreover, imipramine which is an antidepressant was similarly administered in a volume of 3 mL / kg at a dose of 10 mg / kg, and this was used as a positive control group (6 animals). Furthermore, rats fed with water alone served as a negative control group (6 animals). An elevated plus maze test was performed on the rats on the 10th day after administration, and a forced swimming test was performed on the 14th day after administration. On the test day, the test substance was administered 1 hour before the test.

  Hereinafter, the result of the elevated plus maze test is shown in Test Example 1, and the result of the forced swimming test is shown in Test Example 2.

<Test Example 1> (Elevated cross maze test)
The elevated plus maze test is a behavioral test that evaluates anxiety levels. A maze is used in which a road without a wall (open arm) and a road surrounded by a wall (closed arm) cross each other in a cross shape. Usually, the subject animal feels anxiety and fear in an open space without a wall, so that the staying time in the closed arm becomes long. Anti-anxiety medications increase open arm residence time. In this test example, a rat's behavior trajectory is acquired by a video camera installed above the maze, and measured and analyzed by a video tracking system (trade name “CompACT”, manufactured by Muromachi Kikai Co., Ltd.). We calculated the total number of times (momentum) and the percentage of open arm stay time (anxiety level) in both the open and closed arm tracks per minute.

  FIG. 1 shows, as a basic study, when an elevated plus maze test was conducted on a group of rats in which diazepam, an anxiolytic drug, was administered intraperitoneally 30 minutes before the test at a dose of 0.5 mg / kg. The results are shown. As shown in FIG. 1a, the diazepam administration increases the total number of times the open arm and the closed arm enter the runway compared to the control given water only, and the momentum in the maze is increased. I understand that. In addition, as shown in FIG. 1b, it can be seen that administration of diazepam increased the percentage of staying in the open arm compared to the control given water only, and the anxiety level was reduced.

  FIG. 2 shows the results of the elevated plus maze test conducted on the 10th day from the start of administration in the MAK administration group and the imipramine administration group. As shown in FIG. 2a, the MAK administration increases the total number of times the open arm and the closed arm enter the road compared to the control given water only, and as shown in FIG. The percentage of staying time was increased compared to controls given water only. This result was the same tendency as when diazepam was administered. Therefore, it became clear that MAK administration increased the amount of exercise in the maze and reduced the level of anxiety. On the other hand, with imipramine administration, a reduction in anxiety level was observed (FIG. 2b), but no increase in momentum in the maze was observed (FIG. 2a).

<Test Example 2> (Forced swimming test)
The forced swimming test is a method in which a test animal is placed in an inaccessible water tank, and the state of despair (immobility time) caused by the test animal is similar to that of depression in humans. It is considered as a screening method with high specificity for antidepressants because it shortens the kinetic time and is highly correlated with the clinical dose. In this test example, water having a depth of 25 cm was poured into a cylindrical container, a rat was put therein, and the proportion of the immobility time during which the operation was not performed was determined within the observation time of 5 minutes.

  In FIG. 3, as a basic study, when a forced swimming test was performed on a group of rats in which imipramine, an antidepressant, was administered intraperitoneally 30 minutes before the test at a dose of 10 mg / kg or 30 mg / kg. The results are shown. As shown in FIG. 3, it can be seen that immobilization time is reduced depending on the dose of imipramine administration as compared with the control given with physiological saline alone.

  FIG. 4 shows the results of a forced swimming test conducted on the 14th day from the start of administration in the MAK administration group and the imipramine administration group. As shown in FIG. 4, the immobility time was significantly reduced by MAK administration (risk rate <0.05) compared to the control given water alone. A significant reduction in immobility time was also observed with imipramine administration.

(3) Other Preparation Examples of Culture Medium Extract <Preparation Example 2> (Solid culture of Shiitake mushroom using bagasse)
80 parts by mass of bagasse and 20 parts by mass of defatted rice bran were blended, adjusted to a moisture content of 70%, a solid medium was prepared, and autoclaved as usual. This solid medium was inoculated with shiitake mycelia and cultured for 4 months in a culture chamber adjusted to a temperature of 22 to 24 ° C. After the mycelium invaded in the medium, it was transferred to a temperature treatment chamber at 24 to 34 ° C. Warm for hours and then treat in a cold room at 10 ° C. for 5 days. Then, it culture | cultivated for 2 days in the said culture room, and the culture medium was crushed twice with the crusher. The crushed medium was treated at 50 ° C. for 4 hours to promote autolysis, then packed in an extraction tank, and extracted for 16 hours while circulating 60 ° C. warm water. The obtained extract was pre-filtered with a filter cake made of diatomaceous earth, further filtered and sterilized with a membrane filter, concentrated, and freeze-dried to obtain a brown powder. The results of analyzing the components were as follows.

・ Carbohydrate: 35% (phenol sulfate method)
・ Protein: 12% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 35% (acetyl bromide method)
・ Inorganic: 13% (direct ashing method)

<Preparation Example 3> (Solid culture of Shiitake mushroom using corn stalk)
Corn stalks were washed with water, dried, crushed, and rice bran was added at a mass ratio of 20%, adjusted to 70% moisture, a solid medium was prepared, and autoclaved as usual. Shiitake mycelium was inoculated into this solid medium, and a culture extract was obtained in the same manner as in Preparation Example 2. The results of analyzing the components were as follows.

・ Sugar: 40% (phenolic sulfuric acid method)
・ Protein: 12% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 35% (acetyl bromide method)
・ Inorganic: 12% (direct ashing method)

<Preparation Example 4> (Solid culture of N. gonorrhoeae using Kumagusu)
10% by mass of rice bran was added to the crushed fiber residue obtained by extracting the pigment and scent of kumabuchi with ethyl alcohol, and a solid medium was prepared by adjusting the water content to 70%, followed by high-pressure steam sterilization as usual. This solid medium was inoculated with mycelium of mannen koji, and a culture extract was obtained in the same manner as in Preparation Example 2. The results of analyzing the components were as follows.

・ Sugar: 30% (phenolic sulfuric acid method)
・ Protein: 15% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 35% (acetyl bromide method)
・ Inorganic: 10% (direct ashing method)

<Preparation Example 5> (Liquid culture of Shiitake mushroom using bagasse)
To 1 L (liter) of water, 60 g of bagasse finely crushed with a mixer, 20 g of dried okara, 20 g of rice bran, and 2 g of yeast extract were added, followed by high-pressure steam sterilization as usual. This liquid medium is inoculated with mycelia of shiitake mushrooms and aerated for 3 weeks at 23 ° C., and then the culture solution is heated at 60 ° C. for 1 hour and at 80 ° C. for 2 hours to promote the self-digestion reaction of the mycelium enzyme Metabolites were extracted. The culture solution was centrifuged at 3,000 rpm for 30 minutes, the precipitate was removed, and a brown solution was obtained. The extract was lyophilized and analyzed, and the results were as follows.

・ Carbohydrate: 38% (phenol sulfate method)
・ Protein: 16% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 35% (acetyl bromide method)
・ Other: 11%

<Preparation Example 6> (Liquid culture of gonococcus manneum using bear candy)
90 g, 10 g of rice bran, and 5 g of yeast extract were added to 1 L of water and the fiber residue obtained by extracting the pigment residue and fragrance of Kumagusu with ethyl alcohol finely with a mixer was sterilized under high pressure steam as usual. This liquid medium was inoculated with mycelium of mannen koji, and aerated at 23 ° C. for 2 weeks. The culture was heated and centrifuged in the same manner as in Preparation Example 4 to obtain a dark brown liquid. The extract was lyophilized and analyzed, and the results were as follows.

・ Carbohydrate: 33% (phenol sulfate method)
・ Protein: 18% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 40% (acetyl bromide method)
・ Others: 9%

<Preparation Example 7> (Solid culture of Yamabushi persimmon using bagasse)
65 parts by mass of bagasse, 20 parts by mass of wheat bran, 10 parts by mass of rice bran, and 5 parts by mass of dry yeast were mixed, adjusted to a moisture of 70%, a solid medium was prepared, and autoclaved as usual. This solid medium was inoculated with mycelium of Yamabushi mushroom, and a culture extract was obtained in the same manner as in Preparation Example 2. The results of analyzing the components were as follows.

・ Carbohydrate: 43% (phenol sulfuric acid method)
・ Protein: 12% (semi-micro Kjeldahl method)
・ Water-soluble lignin: 28% (acetyl bromide method)
・ Inorganic: 10%

Claims (2)

It is characterized by containing, as an active ingredient, an extract containing saccharides, proteins and water-soluble lignin obtained by culturing mycelium of mannen koji using a medium containing bagasse and rice koji and extracting from the culture. An antidepressant.   The antidepressant according to claim 1, wherein the extract is obtained by performing autolysis of basidiomycetes from the culture.