JP6427638B1 - Composition for prevention, amelioration or treatment of sleep disorder, comprising a fermentation broth of Bacillus subtilis as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention, amelioration or treatment of sleep disorders, which comprises a fermentation broth of Bacillus subtilis as an active ingredient. Thereby, the pharmaceutical composition or the food composition of the present invention is produced by producing polygamma glutamic acid using Bacillus subtilis derived from clean rice, and prevents, ameliorates or treats sleep disorders such as insomnia, There is an effect that it is possible to improve the quality of sleep. [Selected figure] Figure 4

Description

  The present invention relates to a pharmaceutical composition for the prevention or treatment of sleep disorders, and a food composition for the prevention or amelioration of sleep disorders.

  The market size of the insomnia treatment in the US has reached about 2 trillion won, and recently it has shown a growth rate of over 20% every year. A typical US sleep disorder drug is the product Stillox, which has a market share of about 50% in the sleep agent market of not only the US but also Korea. In general, sleep agents work well in short-term use, but long-term use for 4 weeks or more has problems of dependence and side effects, especially for elderly people with a high prevalence of insomnia. You need to be careful about side effects when taking it. Therefore, there is an increasing interest in natural herbal sleep supplements that can replace sleep agents, and the market size is expanding. Currently, there are more than 10 natural sleep aids most commonly used in the United States and European countries, but very few reports have shown the effects of these sleep aids on insomnia treatment through scientific studies It is done. According to a US consumer report, 64% of people who use sleep agents and 20% of people who use sleep aids have experienced the effect of promoting sleep, and It turns out that the usefulness is very low. In the case of Korea, there are currently no Korean natural sleep supplement products, which are health functional foods for promoting and improving sleep recognized by the Food and Drug Safety Agency, and research on related functional ingredients is also limited. From now on, with the increase of elderly people's population, it is predicted that the demand to solve insomnia will increase while reducing medical expenses, and it is required to excavate natural sleep enhancement food materials that can be purchased inexpensively without a doctor's prescription. ing.

  Polygamma glutamic acid has been certified as a health functional ingredient by the Food and Drug Administration. Poly-gamma-glutamic acid has been certified to date as a function of enhancing immunity at a dose of 1000 mg / day. Other functions not known to be certified but known from dissertation include the improvement of obesity, the improvement of hyperlipidemia, and the improvement of intestinal calcium absorption. Studies related to improvement of sleep disorder by intake of polygammaglutamic acid have not been conducted yet.

Korean Patent Application Publication No. 10-0947353 Korean Registered Patent Publication No. 10-1690175

  An object of the present invention is to produce polygamma glutamic acid using Bacillus subtilis derived from Chongqingan, and administering it to prevent or treat sleep disorders such as insomnia and improve sleep quality It is an object of the present invention to provide a pharmaceutical composition that can be

  In addition, another object of the present invention is to produce polygamma glutamic acid using Bacillus subtilis derived from silkworm and apply it as a functional food to prevent or ameliorate sleep disorders such as insomnia, and for sleep It is an object of the present invention to provide a food composition which can improve the quality.

According to one aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating sleep disorders, which comprises a fermentation broth of Bacillus subtilis and magnesium (Mg) as active ingredients.

  The fermentation broth may comprise poly-gamma glutamic acid.

  The average molecular weight of the polygammaglutamic acid may be 1 to 15,000 kDa.

  The pharmaceutical composition may include polygammaglutamic acid and magnesium in a weight ratio of 1:50 to 50: 1.

  The sleep disorder may be one or more selected from insomnia, sleep-related respiratory disorder, sleep phase regression or progressive syndrome, and jet lag syndrome.

According to another aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of sleep disorders, which comprises poly-gamma glutamic acid and magnesium (Mg) as active ingredients.

According to still another aspect of the present invention, there is provided a food composition for preventing or improving sleep disorders, which comprises a fermentation broth of Bacillus subtilis and magnesium (Mg) as active ingredients.

  The fermentation broth may comprise poly-gamma glutamic acid.

  The food composition may further include magnesium (Mg).

According to still another aspect of the present invention, there is provided a food composition for preventing or improving sleep disorders, which comprises poly-gamma glutamic acid and magnesium (Mg) as active ingredients.

  The fermentation of step (b) may be performed at 28-40 ° C.

  After the step (c), the purified product may be further pulverized to control the particle size of the polygammaglutamic acid.

  According to yet another aspect of the present invention, (a) preparing a Bacillus subtilis strain; (b) fermenting the strain with a culture solution to produce a fermentation broth; and (c) The method for producing a food composition for preventing or improving sleep disorder is provided, comprising the steps of: separating and purifying the fermentation broth to obtain a purified product containing polygammaglutamic acid.

  The pharmaceutical composition of the present invention can produce polygamma glutamic acid using Bacillus subtilis derived from the cleanser, and administration thereof can prevent or treat sleep disorders such as insomnia and improve sleep quality The effect is

  In addition, the food composition of the present invention produces polygamma glutamic acid using Bacillus subtilis derived from clean rice and applies it as a functional food to prevent or ameliorate sleep disorders such as insomnia, and for sleep There is an effect that it is possible to improve the quality.

It is a figure showing roughly the design of the clinical study by example 1 of an examination. It is the figure which showed the question to a Pittsburgh sleep questionnaire (Pittsburgh Sleep Quality Index, PSQI). It is a figure showing roughly the design of the animal test by example 2 of an examination. It is the figure which showed the result of the total sleep time by Test example 2, and a wake time. It is a figure showing roughly the design of the animal test by example 3 of an examination. It is the figure which showed the result of the total sleep time by Test example 3, and the awakening time.

  While the invention is susceptible to various modifications and alternative embodiments, the specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. However, it is to be understood that the present invention is not limited to a particular embodiment, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the present invention. In the description of the present invention, when it is determined that specific descriptions of related known techniques may obscure the subject matter of the present invention, the detailed descriptions thereof will be omitted.

  Hereinafter, the pharmaceutical composition for preventing or treating sleep disorders of the present invention will be described.

  The pharmaceutical composition of the present invention comprises a fermentation broth of Bacillus subtilis as an active ingredient.

  Preferably, the Bacillus subtilis may be Bacillus subtilis Chungkook jang deposited under accession number KCTC 0697BP.

  The Bacillus subtilis is a test strain, which can be distributed and used, and in some cases, can be obtained by traditional fermentation of clean rice.

  The fermentation broth is characterized by containing poly-gamma glutamic acid.

  The average molecular weight of the polygammaglutamic acid is preferably 1 to 15,000 kDa, more preferably 100 to 10,000 kDa, and still more preferably 1,000 to 7,000 kDa.

  By increasing the number of glass carboxyl groups in the molecular structure as the molecular weight of poly-gamma-glutamic acid increases, the reactivity with cationic minerals increases, and the role of mineral mediator can be effectively played. . The results show anti-cancer ability only in experimental animals administered with high molecular weight (2,000 kDa) polygammaglutamic acid.

  It is characterized by the molecular weight being very large compared with having the molecular weight of 300-700 kDa of the polygamma glutamic acid manufactured by the conventional Bacillus subtilis natto (Bacillus subtilis Natto).

  The fermentation broth can be subjected to separation and purification steps to remove Bacillus subtilis strains, cultures, and impurities in addition to polygammaglutamic acid.

  The pharmaceutical composition may further comprise magnesium (Mg), in which case polygammaglutamic acid and magnesium may be contained in a weight ratio of 1:50 to 50: 1, preferably 1:10 to 10 The weight ratio may be 1: 1, more preferably 1: 5 to 5: 1.

  The sleep disorder includes insomnia, sleep related breathing disorder, sleep phase regression syndrome or sleep phase progression syndrome, jet lag syndrome, etc., but the scope of the present invention is not limited thereto, and various forms of It may include sleep disorders.

  In addition, the present invention provides a pharmaceutical composition for preventing or treating sleep disorders, which comprises poly-gamma glutamic acid as an active ingredient. The said polygamma glutamic acid is not limited to what can be obtained from the fermentation culture solution of Bacillus subtilis (Bacillus subtilis).

  Hereinafter, the method for producing the pharmaceutical composition for preventing or treating sleep disorders of the present invention will be described.

  First, Bacillus subtilis strains are prepared (step a).

  The Bacillus subtilis is preferably Bacillus subtilis Chungkook jang deposited under accession number KCTC 0697BP.

  The strain is a test strain, which can be used after being distributed and, in some cases, can also be obtained by a traditional fermentation method of pure and clear.

  Next, the strain is fermented with a culture solution to produce a fermentation culture solution (step b).

  The fermentation is preferably performed at 28 to 40 ° C, more preferably 35 to 39 ° C.

  Thereafter, the fermentation broth is separated and purified to obtain a purified product containing polygammaglutamic acid (step c).

  Separation and purification can be carried out by filtration, alcohol precipitation and the like.

  Finally, a grinding step can also be performed to adjust the particle size of polygammaglutamic acid.

  On the other hand, the term "including as an active ingredient" as used herein is meant to include an amount sufficient to achieve the efficacy or activity of polygammaglutamic acid. In one embodiment of the present invention, polygammaglutamic acid in the composition of the present invention is, for example, 0.001 mg / kg or more, preferably 0.1 mg / kg or more, more preferably 1 mg / kg or more, still more preferably 10 mg It contains more than / kg. Since polygammaglutamic acid is a natural product and there is no side effect in the human body even if overdosed, the upper limit of the amount of polygammaglutamic acid contained in the composition of the present invention is selected by the person skilled in the art within a suitable range. Can be implemented.

  The pharmaceutical composition of the present invention can be manufactured using pharmaceutically suitable and physiologically acceptable auxiliaries besides the above-mentioned active ingredients. As the adjuvant, excipients, disintegrants, sweeteners, binders, binders, coatings, expanding agents, lubricants, lubricants, flavors and the like can be used.

  The above-mentioned pharmaceutical composition can be preferably formulated as a pharmaceutical composition containing, in addition to the above-mentioned active ingredients, one or more additional pharmaceutically acceptable carriers for administration.

  The form of the preparation of the pharmaceutical composition is granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, injectable solutions, etc. It is also good. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Also, if desired or necessary, suitable combinations, lubricants, disintegrants and color formers may also be included as a mixture. Suitable binders are, but not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate Sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

  In the composition to be formulated into a liquid solution, as a pharmaceutically acceptable carrier suitable for sterilization and living body, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, malto Dextrin solution, glycerol, ethanol and one or more of these components can be mixed and used, and, if necessary, other common additives such as antioxidants, buffers, bacteriostatic agents and the like are added can do. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form injectable preparations such as aqueous solutions, suspensions, emulsions etc., pills, capsules, granules or tablets. Can be formulated.

  The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc. , Preferably oral administration.

  Suitable dosages of the pharmaceutical composition of the present invention are factors such as formulation method, mode of administration, patient's age, body weight, sex, pathological condition, food, time of administration, route of administration, excretion rate and response sensitivity. However, a routinely skilled physician can readily determine and prescribe the desired therapeutic or prophylactically effective dosage. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 0.001-10 g / kg.

  The pharmaceutical composition of the present invention is formulated by using a pharmaceutically acceptable carrier and / or excipient by a method which can be easily implemented by those skilled in the art to which the present invention belongs. , May be manufactured in unit dose form or in multi-dose containers. At this time, the dosage form may be in the form of a solution, suspension or emulsion in oil or aqueous medium, or may be in the form of extract, powder, granules, tablet or capsule, and dispersion Or may further contain a stabilizer.

  In addition, the present invention provides a method for preventing or treating sleep disorders, which comprises the step of administering the fermentation broth of Bacillus subtilis to a subject.

  The present invention also provides a method for preventing or treating sleep disorders, which comprises the step of administering poly-gamma glutamic acid to a subject.

  In addition, the present invention provides a food composition for preventing or improving sleep disorders, which comprises a fermentation broth of Bacillus subtilis as an active ingredient.

  The Bacillus subtilis is preferably Bacillus subtilis Chungkook jang deposited under accession number KCTC 0697BP.

  The description of the food composition is the same as the description of the pharmaceutical composition for the prevention or treatment of the sleep disorder of the present invention described above, so that the specific content is referred to the part.

  The present invention also provides a food composition for preventing or improving sleep disorder, which comprises poly-gamma glutamic acid as an active ingredient. The said polygamma glutamic acid is not limited to what can be obtained from the fermentation culture solution of Bacillus subtilis (Bacillus subtilis).

  The present invention also provides a method of producing a food composition for preventing or improving sleep disorder.

  The description of the method of producing the food composition is the same as the description of the method of producing the pharmaceutical composition for preventing or treating sleep disorders of the present invention described above, and thus the specific content thereof is referred to.

  The food composition according to the present invention can be used as a functional food or can be added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meats, chocolates, pizzas, breads, ramen, other noodles, gums, ice cream And vitamins and health supplements.

  The food composition of the present invention can contain, as an active ingredient, not only polygammaglutamic acid but also ingredients generally added in the preparation of foods, such as proteins, carbohydrates, fats, nutrients, seasonings and flavors. Containing agents. Examples of carbohydrates mentioned above are monosaccharides such as glucose, fructose etc .; disaccharides such as maltose, sucrose, oligosaccharides etc; and polysaccharides such as conventional sugars such as dextrin, cyclodextrin etc. and xylitol And sugar alcohols such as sorbitol and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin etc.)) and synthetic flavoring agents (saccharin, aspartame etc.) can be used. For example, when the food composition of the present invention is produced as drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts other than polygammaglutamic acid Etc. can be included further.

  The present invention provides a health functional food comprising a food composition for improving or preventing sleep disorders, which comprises polygammaglutamic acid as an active ingredient. A health functional food is a food produced by adding polygamma glutamic acid to food materials such as beverages, teas, spices, gums and confectionery, or produced by encapsulation, powderization, suspension, etc. Ingestion of H means to bring about a specific effect on health, but unlike general medicines, since it uses food as a raw material, it has the advantage that there are no side effects that may occur when taking medicine for a long time. The health functional food of the present invention thus obtained is very useful because it can be taken on a daily basis. The amount of poly-gamma-glutamic acid added to such a health functional food is different depending on the type of the health functional food, and thus can not be defined uniformly, but it is added within a range that does not impair the original taste of the food. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight, based on the target food. In the case of a health functional food which is in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. . In one embodiment, the health functional food of the present invention may be in the form of a pill, a tablet, a capsule or a beverage.

[Example]
Example 1: Production of poly-gamma-glutamic acid powder (1) Fermentation process

  A strain of Bacillus subtilis Chungkookang (KCTC 0697BP) derived from Cinnamon which is a traditional fermented food was prepared.

Basic medium for production (glucose 5%, L-glutamic acid 5%, (NH ₄ ) ₂ SO ₄ 1%, KH ₂ PO ₄ 0.27%, Na ₂ HPO ₄ 12H ₂ O 0.42%, NaCl 0.05% Using a 0.3 L solution of MgSO ₄ · 7 H ₂ O, 1 ml / L of vitamin solution, pH 6.8) and culturing in a 50 L fermenter at 32 ° C, 150 rpm, air injection rate 1 vvm for 24 hours A culture solution was produced.

(2) Separation and Purification Step The fermentation broth prepared in the above fermentation step is subjected to primary filtration with a filter press, followed by purification and concentration using a UF membrane (Ultra-filtration Membrane). Filter press filtration. Next, ethanol precipitation was carried out to remove impurities, and after washing, it was redissolved and finally freeze-dried, and then ground to obtain polygammaglutamic acid powder having an average molecular weight of about 2000 kDa. As the polygamma glutamic acid used in Test Example 1, one produced by the method of Example 1 was used.

[Test example]
Test Example 1: Clinical trial The design of this clinical trial is schematically shown in FIG. 1, and the clinical trial method will be described based on FIG.

In this study, we conducted a randomized, double-blind, placebo-controlled, cross-mix study in adults with a body mass index of at least 23 kg / m ² and an age of at least 20 years. However, those who have chronic diseases (such as chronic diseases such as liver, kidney and cardiovascular disease, cancer, blood related diseases etc.), those who regularly use drugs due to diseases, allergic symptoms when eating pregnant women, food or nutritional supplements Those who experienced it, those who currently take nutritional supplements, those who have anemia and those who donated blood three months ago were excluded.

  The selected subjects are assigned '1' and '2' to 'A' and '1 and 2' to 'B' and are randomly allocated to each test. I did not know if I was assigned. The study was divided into two groups, 'A study' and 'B study' depending on the type of supplement to be evaluated, and group A-1 (placebo group) was a placebo (placebo: corn starch), group A-2 Was administered Mg, the B-1 group (placebo group) was supplemented with Mg, and the B-2 group was supplemented with Mg and γ-PGA. Mg was used as a powder in the form of pills, and was supplemented with 300 mg of Mg daily, and γ-PGA was supplemented with 600 mg daily. Regarding the preparation of questionnaires corresponding to the results of this study (outcome), anthropometry, blood pressure measurement, and blood sampling, a total of 4 times were performed with the same contents before and after the start of the test.

  Questionnaires consist of general items, lifestyle information, sleep quality, etc., and surveys on sleep quality use the Pittsburgh Sleep Quality Index (PSQI), for reference The questions are shown in FIG. The PSQI value indicates that the higher the score, the lower the quality of sleep. Statistical analysis of the PSQI values was performed using SAS software (SAS 9.1.3, SAS Institute, Cary, NC, USA).

  The PSQI questionnaire is a self-administered questionnaire for evaluating the quality of sleep in the past month, and the detail items are subjective sleep quality, sleep latency, sleep It consists of time (sleep duration), habitual sleep efficiency, sleep disorder, sleep disorder, use of sleep medication, daytime dysfunction, and each item has 0 points. The total score is 21 points from 3 points. The item 'subjective sleep quality' among the above-mentioned detail items means that the higher the score, the lower the subjective sleep quality. That is, a group having a PSQI global score of 5 or less is classified as a satisfactory sleep group (good sleeper), and a group having 6 or more points is classified as an unsatisfactory sleep group (poor sleeper). Also, it is pointed out in most of the papers that there are no significant problems with PSQI reliability [Reference: Joo NS. Koran J Fam Med. 2010; 31: 745-746].

  In order to evaluate the validity of the PSQI question which is a subjective sleep evaluation method, sensitivity and specificity of PSQI were evaluated for 80 insomnia patients and 45 healthy people. As a result, sensitivity was 98.7 and specificity was 84.4 when diagnosing insomnia based on a total of 6 points (satisfied sleep) of PSQI questions. This indicates that the PSQI question is reliable as a diagnosis of insomnia [Reference: Backhaus J et al. J Psychosom Res. 2002; 53: 737-740].

  The results of the clinical trials conducted in this way are shown in Table 1 below.

  In this study, group B-2 (Mg + γ-PGA supplementation) had a lower PSQI score after intake of the nutrient than before intake of the nutrient, while the Mg supplementation group had a PSQI score before and after intake of the nutrient. The score shows that there is almost no change. (P <0.05)

Test Example 2 Animal Test for Observation of Effects of γ-PGA and Mg Supplement The design of the present animal test is schematically shown in FIG. 3, and the animal test method will be described based on FIG.

  4 groups of 20 white rats whose sleep was restricted by intake of caffeine (20 mg / kg) (comparative group, group supplemented with 25 mg / kg polygammaglutamic acid, group supplemented with 300 mg / kg magnesium, 25 mg / kg polygammaglutamic acid + magnesium The same number was assigned to 300 mg / kg (supplemented group), and after oral administration for 2 weeks, the effect of supplementation with polygammaglutamic acid and magnesium was evaluated by an electroencephalographic experiment. The result of the total sleep time and awakening time by this is shown in FIG. According to this, it is shown that the total sleep time increased and the awakening time decreased as compared with the other groups in the group supplemented with 25 mg / kg of polygammaglutamic acid and 300 mg / kg of magnesium, and the supplementation with polygammaglutamic acid and magnesium Has shown that it can help sleep.

Test Example 3: Animal test for observation of the effect of a dose of γ-PGA The design of this animal test is schematically shown in FIG. 5, and the clinical test method will be described with reference to FIG.

  In mice that have been sleep-restricted by taking caffeine (20 mg / kg), after oral administration of 25 mg / kg of polygammaglutamic acid to 10 mice, 5 of them are 25 mg / kg and the remaining 5 are 25 Oral administration of 50 mg / kg for 1 week, the effect of the replacement dose of polygammaglutamic acid was evaluated by an electroencephalographic experiment. The results of the total sleep time and awake time from the electroencephalogram measurement experiment by this are shown in FIG. According to this, it is shown that the total sleep time increased and the awakening time decreased as compared with the 25 mg / kg supplementation group in the polygammaglutamic acid 50 mg / kg replacement group, and the polygammaglutamic acid supplement content was high. It can be seen that the more effective the increase in sleep time and the decrease in arousal time.

  While the embodiments of the present invention have been described above, those having ordinary skill in the art can add components without departing from the concept of the present invention described in the claims. The present invention can be variously modified and changed by changes, deletions, additions, and the like, which are also included in the scope of the present invention.

Claims (13)

A pharmaceutical composition for the prevention or treatment of sleep disorders, which comprises a fermentation broth of Bacillus subtilis and magnesium (Mg) as active ingredients.   The pharmaceutical composition for preventing or treating sleep disorders according to claim 1, wherein the fermentation broth comprises poly-gamma glutamic acid. The pharmaceutical composition for preventing or treating sleep disorders according to claim 2 , wherein the average molecular weight of the polygammaglutamic acid is 1 to 15,000 kDa. The pharmaceutical composition for preventing or treating sleep disorders according to claim 1 , wherein the pharmaceutical composition comprises polygammaglutamic acid and magnesium in a weight ratio of 1:50 to 50: 1.   The prevention of a sleep disorder according to claim 1, wherein the sleep disorder is at least one selected from insomnia, sleep related breathing disorder, sleep phase regression or progressive syndrome, and jet lag syndrome. Therapeutic pharmaceutical composition. Pharmaceutical composition for prevention or treatment of sleep disorder which contains poly gamma glutamic acid (poly-gamma glutamic acid) and magnesium (Mg) as an active ingredient. A food composition for preventing or improving sleep disorders, which comprises a fermentation broth of Bacillus subtilis and magnesium (Mg) as active ingredients. The food composition for preventing or improving sleep disorders according to claim 7 , wherein the fermentation broth comprises poly-gamma glutamic acid. A food composition for preventing or improving sleep disorders, which comprises poly-gamma glutamic acid and magnesium (Mg) as active ingredients. (A) preparing a Bacillus subtilis strain;
(B) fermenting the strain with a culture solution to produce a fermentation culture solution;
(C) separating and purifying the fermentation broth to obtain a purified product containing polygammaglutamic acid, and producing a pharmaceutical composition for preventing or treating sleep disorder. The method for producing a pharmaceutical composition for preventing or treating sleep disorders according to claim 10 , wherein the fermentation in step (b) is performed at 28 to 40 ° C. The pharmaceutical composition for preventing or treating sleep disorders according to claim 10 , further comprising the step of crushing the purified product to adjust the particle size of the polygammaglutamic acid after the step (c). Method of making the composition. (A) preparing a Bacillus subtilis strain;
(B) fermenting the strain with a culture solution to produce a fermentation culture solution;
(C) separating and purifying the fermentation broth to obtain a purified product containing polygammaglutamic acid, and producing a food composition for preventing or improving sleep disorder.