KR102207581B1 - Pharmacological Composition Comprising Lactic acid Bacteria Fermented Product of Morus alba Extract for Prevention or Treatment of Depressive Disorder or Anxiety Disorder - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing the lactic acid bacteria fermentation broth of the heartworm extract as an active ingredient, wherein the composition is fermented with lactic acid bacteria extracts from which the active component of heartbreak is extracted through hot water extraction and ultrasonic extraction to prevent depression or anxiety disorders or It shows excellent effect in treatment, and can reduce the occurrence of side effects caused by existing treatments for depression or anxiety disorders.

Description

Pharmaceutical Composition Comprising Lactic acid Bacteria Fermented Product of Morus alba Extract for Prevention or Treatment of Depressive Disorder or Anxiety Disorder}

The present invention relates to a pharmaceutical composition for the prevention or treatment of depression or anxiety disorders, containing a fermented broth of lactic acid bacteria extract from the heart of the heart as an active ingredient.

Looking at the proportion of the aging population in Korea, in 2000, the proportion of the population aged 65 or older reached 7.2% of the total population, entering an aging society, and in 2019, this ratio is expected to exceed 14% to enter the aging society. have. Due to the increase in the aging population, chronic degenerative diseases have emerged as a bigger problem than acute infectious diseases that have been the main cause of death for the past 50 years.

Alzheimer, a representative disease of degenerative brain diseases, is the most common form of dementia, and neurons in the brain are degenerated and the brain is contracted, causing progressive mental disorders. As representative dementia treatments known so far, tacrine (Cognex, 1994) and donepezil (Aricept, 1996) have been approved and used by the US FDA. The mechanism of action of these drugs is to increase the concentration of the neurotransmitter acetylcholine by inhibiting the activity of the enzyme acetylcholinesterase (AChE), an enzyme that degrades acetylcholine, which plays a central role in the central nervous system. It is known to prevent and treat. However, the tacrine is expensive compared to its efficacy and has serious hepatotoxicity problems, and the done cortex does not have hepatotoxicity, but is known to have various side effects such as vomiting, nausea, and diarrhea by stimulating the parasympathetic nerve.

In addition, patients with dementia have behavioral and psychological symptoms of dementia (BPSD), and the main symptoms are agitation, anxiety, depression, hallucinations, and personality changes. personality change). Accordingly, dementia treatment, antidepressants, and anti-anxiety drugs are being prescribed together in the treatment of dementia, but memory is reduced due to side effects of the complex prescription, and BPSD that does not respond to drugs is a problem.

Accordingly, the present inventors searched for a new composition derived from a natural substance capable of improving depression or anxiety disorders while having few side effects caused by drugs, and as a result, Sangsimja (Od), a medicinal material verified in oriental medicine through the lactic acid bacteria fermentation method. A composition that can be applied to prevent, ameliorate, or treat depression or anxiety disorders by containing a useful bioactive substance extracted from was developed.

Korean Patent Registration No. 10-1937321

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of depression or anxiety disorders, containing the fermented broth of lactic acid bacteria extract of the extract as an active ingredient.

Here, the upper heart extract may be one containing the active ingredient extracted from the upper heart using hot water extraction and ultrasonic extraction.

The lactic acid bacteria may be Lactobacillus brevis DF01 or Pediococcus acidilactici K10.

The fermentation broth may be cultured by inoculating 0.1 to 5 parts by weight of lactic acid bacteria to 100 parts by weight of the upper heart extract.

In addition, the present invention (a) preparing by crushing the heartache; (b) extracting the pulverized upper core at 80 to 120° C. with hot water for 1 to 5 hours to obtain a first extract; (c) ultrasonically extracting the first extract at 40 to 80° C. for 10 to 60 minutes to obtain a second extract; And (d) inoculating the second extract with lactic acid bacteria and fermenting at 25 to 45° C. for 2 to 5 days to obtain a fermentation broth.

Here, the step (b) may be to extract hot water by mixing the pulverized upper core and water in a ratio of 1:5 to 20 (w/v).

The step (c) may be performing ultrasonic treatment after adjusting the pH of the first extract to 6 to 7.

The lactic acid bacteria in step (d) may be Lactobacillus brevis DF01 or Pediococcus acidilactici K10.

In the step (d), the initial inoculation concentration of lactic acid bacteria may be 10 ⁷ to 10 ⁹ cfu/mL.

After the step (d), the step of filtering the fermentation broth of step (d) may be further included.

<Pharmaceutical composition for preventing or treating depression or anxiety disorder>

The composition according to an embodiment of the present invention may be a pharmaceutical composition for the prevention or treatment of depression or anxiety disorders containing a fermented broth of lactic acid bacteria extract of the heartwort extract as an active ingredient.

Sangsimja (Audi) is the fruit of Morus alba L. , a deciduous tree belonging to the Moraceae family , and contains fatty acids such as sugar, tannic acid, vitamins, and linolic acid. It is known to contain. These heartaches are collected when they change from blue to purple and black as they ripen, and are known to be effective in dizziness, tinnitus, googal, sogal, tonic, pain reliever, insomnia, low back pain, constipation, etc., and can be used for edible and medicinal purposes. Republic of Korea Patent Laid-Open No. 10-2005-0067131 relates to a composition containing a heartworm extract having monoamine oxidase (MAO) inhibitory activity, and the above heartbreak extract is also used in diseases such as depression and Parkinson's disease related to MAO enzyme. It is disclosed that it is effective. Therefore, the present inventors are effective in the lactic acid bacterium fermentation broth of the extract of lactic acid bacteria having a large effect on improving depression or anxiety disorder by applying the lactic acid bacteria fermentation method in order to select the lactic acid bacteria fermentation method in order to more efficiently use the active ingredients contained in the sedation and anxiety disorder It was possible to obtain a pharmaceutical composition for the prevention or treatment of depression or anxiety disorder containing as an ingredient.

In the present specification, the "broken heart extract" includes a useful physiologically active substance or an active ingredient (active ingredient) isolated from the heartworm, and when a solvent is used for extraction, it includes a solvent together with the active ingredient separated from the heartworm it means. The heartache may be raw raw material, dry, or a mixture thereof, and may be finely ground so that the active ingredient can be efficiently extracted. In addition, the upper heart extract also includes an extract that has undergone a conventional purification process. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through the purification method may also be included in the extract. In addition, the extract may be in the form of a powder by an additional process such as distillation under reduced pressure, freeze drying, spray drying, and the like.

The above extract can be obtained using an extraction method known in the art without limitation. Examples of the extraction method include cold needle extraction, ultrasonic extraction, reflux cooling extraction, hot water extraction, pressure extraction, solvent extraction, supercritical extraction, ultrasonic extraction, and the like. When a solvent is used for extraction, an extraction solvent known in the art may be used without limitation. Examples of the extraction solvent include purified water, distilled water, C ₁ ~ C ₄ alcohol, acetic acid, dichloromethane, dimethyl formamide, dimethyl sulfoxide (DMSO), acetone, acetonitrile, ethyl acetate, Methyl acetate, pentane, hexane, chloroform, diethyl ether, carbon tetrachloride, tetrahydrofuran (THF), and the like. Examples of the C ₁ to C ₄ alcohol include methanol, ethanol, propanol, n-butanol, and iso-butanol.

In the present invention, it is preferable to further perform ultrasonic extraction after hot water extraction of the heartache so that the active ingredient of the heartache can be extracted as much as possible. The hot water extraction is useful for extracting water-soluble substances, and when water or alcohol of C ₁ ~ C ₄ is used as a solvent, it is simpler and takes less time than when an organic solvent is used, and the possibility of causing toxicity by the solvent can be reduced. have. The ultrasonic extraction may increase extraction efficiency by destroying solids by the impact effect of ultrasonic energy. The supernatant extract obtained by this method may include an extract obtained by an extraction treatment, a dilution or concentrate of the extract, a dried product obtained by drying the extract, and any one of these preparations or purified products.

Lactobacillus (lactic acid bacteria) refers to total bacteria that break down carbohydrates into lactic acid during metabolic processes. Representative genus belonging to lactic acid bacteria include Lactobacillus sp. , Streptococcus sp. , Pediococcus sp. , Leuconostoc sp. , and Bifidobak It may be the genus Bifidobacterium sp .

In the present invention, it may be Lactobacillus and/or Pediococcus that is easy to handle and has excellent fermentation efficiency of lactic acid bacteria, and in particular, lactobacillus, a kimchi lactic acid bacteria that is more effective in fermentation of vegetable substances than lactic acid bacteria derived from animal foods isolated from yogurt Bacillus brevis DF01 ( Lactobacillus brevis DF01) or Pediococcus acidilactici K10 ( Pediococcus acidilactici K10) is preferred. The fermentation process using such lactic acid bacteria has the advantage of making it easier to extract the active ingredient compared to general biological fermentation or extraction methods using a solvent.

In the present specification, "fermentation" means a process of decomposing sugars in which energy is obtained without the use of oxygen in a narrow sense, and in a broad sense means a process of obtaining substances useful to humans by using microorganisms or fungi. In addition, "fermentation liquid" refers to a material that is beneficial to humans, which is produced through various enzyme actions by culturing lactic acid bacteria in a material to be fermented, and refers to a result obtained by culturing the above-mentioned lactic acid bacteria extract. The fermentation broth is obtained by inoculating the lactic acid bacteria in the lactic acid bacteria extract and culturing the lactic acid bacteria under optimal growth conditions, and in order to improve fermentation efficiency, it is preferable to inoculate 0.1 to 5 parts by weight of lactic acid bacteria in 100 parts by weight of the lactic acid bacteria extract.

The pharmaceutical composition containing the lactic acid bacterium fermentation broth of the extract according to the present invention as an active ingredient may further include a pharmaceutically acceptable carrier. In the above, "pharmaceutically acceptable" refers to a composition that is physiologically acceptable and does not cause allergic reactions or similar reactions such as gastrointestinal disorders and dizziness when administered to humans. Pharmaceutically acceptable carriers include, for example, a carrier for oral administration such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like, and a carrier for parenteral administration such as water, suitable oils, saline, aqueous glucose and glycol. And the like, and may further include a stabilizer and a preservative. Suitable stabilizers may be sodium hydrogen sulfite, sodium sulfite, ascorbic acid and the like. Suitable preservatives may be benzalkonium chloride, methyl-paraben, propyl-paraben, chlorobutanol, and the like.

The pharmaceutical composition may be formulated in a suitable form according to a method known in the art together with a pharmaceutically acceptable carrier as described above. That is, the pharmaceutical composition of the present invention may be prepared in various parenteral or oral administration forms according to known methods. A typical formulation for parenteral administration is a formulation for injection, and an isotonic aqueous solution or suspension is preferred. Formulations for injection can be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component can be formulated for injection by dissolving it in saline or buffer. In addition, the formulation for oral administration may be a powder, granules, tablets, pills and capsules.

The pharmaceutical composition formulated by the above method may be administered in an effective amount through various routes including oral, transdermal, subcutaneous, intravenous, or intramuscular. In the above,'effective amount' refers to an amount that exhibits a preventive or therapeutic effect when administered to a patient. The dosage of the pharmaceutical composition may be appropriately selected according to the route of administration, the subject of administration, age, gender, weight, individual difference, and disease state.

According to an embodiment of the present invention, when the lactic acid bacterium fermentation broth of the heartworm extract is administered to an animal model of depression or anxiety disorder, a forced swim test, a tail suspension test, an open field test ( open field test), the elevated plus maze test (elevated plus maze test), since the immobility time and activity ability can be restored to normal levels, the pharmaceutical composition containing the lactic acid bacteria fermentation broth of the upper heart extract may be used for depression or It can be usefully used for preventing or treating anxiety disorders.

<Method of preparing a pharmaceutical composition for preventing or treating depression or anxiety disorder>

The method for preparing a pharmaceutical composition for preventing or treating depression or anxiety disorder comprises the steps of: (a) preparing a broken heart by crushing it; (b) extracting the pulverized upper core at 80 to 120° C. with hot water for 1 to 5 hours to obtain a first extract; (c) ultrasonically extracting the first extract at 40 to 80° C. for 10 to 60 minutes to obtain a second extract; And (d) inoculating the second extract with lactic acid bacteria and fermenting at 25 to 45° C. for 2 to 5 days to obtain a fermentation broth.

Looking specifically at the steps (a) to (d) are as follows.

(a) preparing by crushing the heartache

The heartache may be in a raw or dry state, or a mixture thereof. In order to efficiently extract the active ingredient from these heartaches, it is prepared by grinding.

Grinding methods known in the art can be used without limitation as a method of crushing the upper core, such as a ball mill, hammer mill, rod mill, vibration mill, roll crusher, centrifugal impact mill, vertical bead mill, abrasion mill, vibration mill, It can be pulverized using a pin mill, hammer mill, pulverizer, crusher, knife cutter, or the like. In the present invention, it can be pulverized for 5 to 30 minutes so that the upper core has uniform particles.

(b) extracting the pulverized upper core with hot water to obtain a primary extract

By hot water extraction of the pulverized upper core, a primary extract containing the active ingredient of the upper core can be obtained.

The method of hot-water extraction of the crushed heartwort is to use water as an extraction solvent to prevent toxic or unnecessary substances from being generated during the extraction process due to the remaining organic solvents in the extract or fermentation broth at a later stage. desirable. In the present invention, it is preferable to extract hot water at 80 to 120°C for 1 to 5 hours, and at this time, hot water extraction by mixing the pulverized heart core and water in a ratio of 1:5 to 20 (w/v) can do. At this time, if the extraction condition is less than 80°C or less than 1 hour, the active ingredient may not be properly extracted from the heartache, and if it exceeds 120°C or 5 hours, the active ingredient may be destroyed or lost by heat. In addition, if the ratio of the heartbreak to water is less than 1: 5, the active ingredient of the heartbreaker cannot be sufficiently extracted due to insufficient solvent, and when it exceeds 1:20, the extracted active ingredient is diluted in the solvent, resulting in lower extraction efficiency. A further step of concentrating may be required.

(c) ultrasonically extracting the first extract to obtain a second extract

The first extract may be subjected to ultrasonic treatment to obtain a second extract.

The primary extract is a state in which a solvent containing an active ingredient extracted from the upper core and a solid upper core are mixed. In order to structurally stabilize the active ingredient in the solvent and prevent the active ingredient from being destroyed or lost due to heat during ultrasonic treatment, the acidity of the primary extract may be adjusted. At this time, it is preferable to adjust the pH of the first extract to 6 to 7 by adding an acidic or basic substance to the first extract. As the acidic or basic substance, substances known in the art may be used without limitation.

The extraction method using ultrasonic waves is a method of extracting using energy generated by ultrasonic vibration. Ultrasound can destroy the insoluble solvent contained in the heart in a water-soluble solvent, and due to the high local temperature generated at this time, the kinetic energy of the reactant particles located around it increases, so that sufficient energy required for the reaction is obtained. By inducing a high pressure due to the impact effect of ultrasonic energy, it can increase the extraction efficiency by enhancing the mixing effect of the material and solvent contained in the heartache. In the present invention, it is preferable to ultrasonically extract the first extract at 40 to 80° C. for 10 to 60 minutes. At this time, when the extraction conditions are less than 40°C or less than 10 minutes, the extraction efficiency may be low, and when the extraction conditions exceed 80°C or 30 minutes, the active ingredient may be destroyed or lost due to heat generated by ultrasonic waves.

(d) inoculating and fermenting the lactic acid bacteria in the secondary extract to obtain a fermentation broth

By fermenting the second extract with lactic acid bacteria, it is possible to obtain a fermentation broth containing all of the active ingredients of the heartworm and the products produced from the lactic acid bacteria using the same.

The secondary extract is in a state in which the active ingredient of the heartache is sufficiently eluted through the steps of hot water extraction and ultrasonic extraction. By fermenting the active ingredient of the heartworm into lactic acid bacteria, it can be converted into a form that is more easily absorbed or used in the human body, or a new useful substance can be created through fermentation.

As a method of fermenting lactic acid bacteria, a method of fermenting lactic acid bacteria known in the art may be used without limitation. In the present invention, in order to provide the optimum culture conditions for lactic acid bacteria to grow, it is preferable to inoculate the lactic acid bacteria in the second extract and ferment at 25 to 45° C. for 2 to 5 days, and in particular, ferment at 30 to 40° C. It is more preferable. At this time, if the fermentation temperature is out of the above range, the fermentation rate may be slowed down or unwanted fermentation products may be generated due to the killing of lactic acid bacteria, and if the fermentation time is out of the above range, the fermentation efficiency decreases or the raw materials are spoiled. Symptoms may occur. Since the lactic acid bacteria fermentation can be cultured under both anaerobic and aerobic conditions, both anaerobic and aerobic fermentation are possible. In addition, in order to improve the growth of lactic acid bacteria, the secondary extract may be maintained at a pH of 4 to 4.5. In addition, in order to promote fermentation by lactic acid bacteria without significantly changing the efficacy of the secondary extract, a carbon source and an energy source of lactic acid bacteria may be added to the secondary extract before inoculation and fermentation of lactic acid bacteria. At this time, it may be added by adjusting the type and amount of the carbon source and energy source in consideration of the fermentation time and the degree of fermentation. As the carbon source and energy source, materials known in the art may be used without limitation, and for example, oligosaccharides, lactose, glucose, fructose, sugar, and mixtures thereof may be used.

The lactic acid bacteria is Lactobacillus genus (Lactobacillus sp.), Streptococcus genus (Streptococcus sp.), Phedi O Rhodococcus genus (Pediococcus sp.), Flow Pocono stock in (Leuconostoc sp.), Bifidobacterium (Bifidobacterium sp . ) Etc. The lactic acid bacteria are preferably Lactobacillus brevis DF01 or Pediococcus acidilactici K10 having excellent fermentation efficiency, and more preferably Pediococcus acidilactici K10.

The initial inoculation concentration of the lactic acid bacteria is 10 ⁷ to 10 ⁹ cfu/mL, and can be fermented by inoculating 0.1 to 5 parts by weight of lactic acid bacteria to 100 parts by weight of the secondary extract. At this time, when the initial concentration and inoculation amount of lactic acid bacteria are out of the above range, fermentation may not be properly performed, and manufacturing cost may increase.

The fermentation broth obtained through such lactic acid bacteria fermentation contains solids such as heartworm residue, lactic acid bacteria, and fermentation impurities, and may form sediments or suspended solids, and may negatively affect the final product. Therefore, in order to remove the solids contained in the fermentation broth, the step of filtering the fermentation broth may be further performed. The method of filtering the fermentation broth may be used without limitation, a filtration method known in the art, for example, it may be filtered using a filter press, a centrifuge, filter paper, or the like. In order to completely remove the solids, the filtration process may be repeated one or more times.

In addition, the fermentation broth may be dried to facilitate storage and use of the fermentation broth. As a method of drying the fermentation broth, a drying method known in the art can be used without limitation. For example, freeze drying, vacuum drying, aeration drying, blow drying, hot air drying, flow drying, spray drying, infrared drying, high frequency drying, etc. have.

The composition containing the lactic acid bacterium fermentation broth of the heartworm extract according to the present invention exhibits excellent effect in preventing or treating depression or anxiety disorder by fermenting the extract from which the active ingredient of the heartworm is extracted through hot water extraction and ultrasonic extraction as lactic acid bacteria. It can reduce the incidence of side effects caused by treatment or anxiety disorder treatment.

1 is a graph showing the result of a forced swimming test in a stressed animal model in which a lactic acid bacteria fermentation broth of an extract of heartworm was administered for 30 days as a result of Example 2-2 according to the present invention.
FIG. 2 is a graph showing the result of Example 2-2 according to the present invention, a result of a forced swimming test in a stressed animal model in which the lactic acid bacteria fermentation broth of the extract of heartworm was administered for 60 days.
3 is a graph showing the result of performing a tail suspension test in a stressed animal model in which the lactic acid bacteria fermentation broth of the extract of Ssangsimja was administered for 30 days as a result of Example 2-2 according to the present invention.
4 is a graph showing the result of performing a tail suspension test in a stressed animal model in which the lactic acid bacteria fermentation broth of the extract of Ssangsimja was administered as a result of Example 2-2 according to the present invention.
5 is a photograph and graph showing the result of performing an open field test in a stress animal model in which a lactic acid bacterium fermentation broth of a heartworm extract was administered for 60 days as a result of Example 2-3 according to the present invention. to be.
6 is a result of Example 2-3 according to the present invention, showing the result of performing an elevated plus maze test in a stressed animal model in which the lactic acid bacteria fermentation broth of the heart extract was administered for 60 days. It is a graph.

Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1: Preparation of fermented extract of heartworm

Morus alba was purchased and used by Taewondang Pharmaceutical Company (Jung-gu, Daegu). After pulverizing the heartwort for 10 minutes or more with a blender, extract it for 3 hours in a constant temperature water bath at 100℃ at 1:10 w/v with distilled water, and then use a sonicator (ultrasonic output 40 kHz) to 60 Further extraction was performed at °C for 30 minutes. The extract was adjusted to pH 6.5 with 1N NaOH, and then Lactobacillus brevis DF01 (source: Korea P&P Research Institute, Yesan) or Pediococcus acidilactici K10 (source: Korea Food Science Research Institute, Gyeonggi-do) at 1 x 10 ⁸ cfu/mL. A 1% inoculation was performed at a concentration and fermented at 37° C. for 3 days, and the lactic acid bacteria fermentation broth of the extract of the upper heart extract from which the remaining bacteria and foreign substances were removed through a filter was used for later experiments.

Example 2: Confirmation of the effect of fermented extract of Sangsimja

Example 2-1. Stress model

The chronic mild stress (hereinafter referred to as'CMS') model was implemented by partially modifying the experimental method based on the experimental model proposed by Willner (1984, Psychopharmacology. 83:1-16.). CMS was performed for a total of 3 weeks. After the completion of CMS, as a behavior test, a forced swimming test, tail suspension test, open field test, and elevated cross maze test were performed.

8-week-old male mice weighing 22 to 24 g (BALB/c; Daehan Biolink, Eumseong, Korea) were divided into 6 groups as follows.

-Normal group (N): non-stress group with daily administration of distilled water (number of subjects: 10)

-Control group (S): stress group administered with distilled water daily (number of individuals: 10)

-Positive control group (F): stress group administered orally for 30 or 60 days with fluoxetine hydrochloride (Sigma), an antidepressant, at 10 mg/kg/day (number of subjects: 10)

-Lactobacillus (L): Stress group in which Duolac Oriental lactobacilli 1 x 10 ⁸ cfu/g was administered orally for 30 or 60 days (Number of individuals: 10)

-Lactobacillus K10 fermentation broth (K10): Stress group in which lactic acid bacterium fermentation broth of the extract of Ssangsimja was orally administered for 30 or 60 days at a concentration of 6.6 mL/kg/day (Number of subjects: 5)

-Lactobacillus DF01 fermentation broth (DF01): Stress group in which lactic acid bacterium fermentation broth from the extract of Ssangsimja was orally administered for 30 or 60 days at a concentration of 6.6 mL/kg/day (Number of individuals: 5)

Mice in the group were randomly assigned and applied so as not to predict various stresses such as restraint, fasting, and isolation for 3 weeks. The normal group was reared in another breeding room to avoid contact with the stressed group, and water and food were supplied freely. After applying the stress, the forced swimming experiment, tail suspension experiment, open field experiment and elevated cross maze experiment were performed.

2-2. Confirmation of antidepressant effect of fermented Sangsimja extract

① Forced swimming test (FST)

After 30 or 60 days of drug administration, the degree of depression of the mice was confirmed by a forced swimming test.

A 50x20x30cm cylindrical plastic equipment was filled with water of 25±1℃ to a height of 23cm, and the mice belonging to the six groups were forced to swim for 6 minutes to observe the behavior of the mice. The behavioral observation video was taken using a video camera connected to the software (Smart Junior, Panlab SL, Barcelona, Spain). The case where the mouse has foot motion for swimming under the water and no motion with only the head exposed above the water was defined as activity/immobility, respectively. The mouse's climbing time, swimming time, and immobility time were automatically measured using software linked with video tracking recording, and the results are shown in FIGS. 1 and 2.

As shown in Figures 1 and 2, when the lactic acid bacteria K10 fermentation broth (K10) or lactic acid bacteria DF01 fermentation broth (DF01) was administered, it was confirmed that the climbing time and swimming time increased and the immobility time decreased compared to the control group (S). In addition, when lactic acid bacteria K10 fermentation broth (K10) or lactic acid bacteria DF01 fermentation broth (DF01) was administered for 60 days (FIG. 2), it was confirmed that swimming time increased and immobility time decreased compared to when administered for 30 days (FIG. 1). Could In particular, it was found that the behavioral improvement effect was improved when the lactic acid bacteria DF01 fermentation broth (DF01) was administered compared to the case where the lactic acid bacteria K10 fermentation broth (K10) was administered.

② Tail suspension test (TST)

The degree of depression of mice 30 or 60 days after drug administration was confirmed through tail suspension experiment.

The tails of mice belonging to the six groups were hung on the top of a 30x30x50cm plastic square device, and the behavior was observed for 6 minutes in a state where noise was blocked under red lighting. The behavioral observation video was taken using a video camera connected to the software (Smart Junior, Panlab SL, Barcelona, Spain). A state in which the mouse is suspended and completely stopped without any movement was regarded as a floating state. During the experiment, the mouse holding the tail and rising was excluded from the experimental results, and the results are shown in FIGS. 3 and 4.

As shown in FIGS. 3 and 4, when the lactic acid bacteria K10 fermentation broth (K10) or the lactic acid bacteria DF01 fermentation broth (DF01) was administered, it was confirmed that the immobility time decreased compared to the control group (S). In particular, it was found that when lactic acid bacteria DF01 fermentation broth (DF01) was administered, the behavior improvement effect was improved compared to when lactic acid bacteria K10 fermented broth (K10) was administered, and the behavior improvement effect was excellent compared to the case where an existing antidepressant (F) was administered. Could In addition, it was found that even if the lactic acid bacteria K10 fermentation broth (K10) or the lactic acid bacteria DF01 fermentation broth (DF01) was administered for 30 days, the behavior improvement effect was excellent.

2-3. Confirmation of anti-anxiety effect of fermented Sangsimja extract

① Open field test

60 days after drug administration, the degree of anxiety of the mice was confirmed through an open field experiment. The open field experiment is a qualitative and quantitative measure of general gait activity and exploration willingness commonly used in rodents.

Mice belonging to the six groups were placed in a 30x40x30cm plastic square box and observed for 5 minutes per mouse. The illumination of the chamber was adjusted to 7-8 lux. Mice were placed in the same environment 30 minutes prior to exposure to the open range. Each mouse was individually placed in the center of the open range and movement was recorded for 5 minutes. The horizontal movement activity was determined through the movement distance of the mouse. The internal 25% range was judged as the central part. The behavioral observation video was taken using a video camera connected to the software (Smart Junior, Panlab SL, Barcelona, Spain). Thereafter, the distance the mouse moved and the time spent in the center were measured every minute through video file analysis using video tracking software, and the results are shown in FIG. 5.

As shown in Figure 5, when lactic acid bacteria K10 fermentation broth (K10) is administered, the central distance increases compared to the control group (S), and when lactic acid bacteria DF01 fermentation broth (DF01) is administered, the total distance and It was confirmed that the peripheral distance increased.

② Elevated plus maze test

After 60 days of drug administration, the degree of anxiety of the mice was confirmed through an expensive cross maze experiment.

The elevated cross maze device consists of two open arms (30x5cm) and two closed arms (5x30cm, wall 15cm high) extending from a common central platform (5x5cm). Became. The structure was formed in the shape of a plus-sign in which corresponding branches were arranged to face each other. The device is raised 50 cm above the floor level.

Mice belonging to the six groups were placed in the elevated cross maze device and observed for 7 minutes per mouse. Each mouse was individually positioned in the center of the open range, and the time spent in each open branch and closed branch was measured. The behavioral observation video was taken using a video camera connected to the software (Smart Junior, Panlab SL, Barcelona, Spain). Thereafter, the time the mouse stayed was measured every second through video file analysis using video tracking software, and the results are shown in FIG. 6.

As shown in Figure 6, when the lactic acid bacteria K10 fermentation broth (K10) or lactic acid bacteria DF01 fermentation broth (DF01) was administered, it was confirmed that the staying time in the open and closed branches increased compared to the control (S). In particular, it was found that the case of administering the lactic acid bacteria K10 fermentation broth (K10) or the lactic acid bacteria DF01 fermentation broth (DF01) was superior to the case where the conventional antidepressant (F) was administered, and the behavior improvement effect was excellent.

Claims (10)

Contains the lactic acid bacteria fermented broth from the extract of the upper heart japonica as an active ingredient,
The Sangsimja extract contains the active ingredient extracted from the Sangsimja using hot water extraction and ultrasonic extraction,
The lactobacillus is Lactobacillus brevis DF01 ( Lactobacillus brevis DF01) or Pediococcus acidilactici K10 ( Pediococcus acidilactici K10) is a pharmaceutical composition for the prevention or treatment of depression or anxiety disorders.
delete delete The method of claim 1,
The fermentation broth is a pharmaceutical composition that is cultured by inoculating 0.1 to 5 parts by weight of lactic acid bacteria to 100 parts by weight of the upper heart extract.
(a) preparing by crushing the heartache;
(b) extracting the pulverized upper core at 80 to 120° C. with hot water for 1 to 5 hours to obtain a first extract;
(c) ultrasonically extracting the first extract at 40 to 80° C. for 10 to 60 minutes to obtain a second extract; And
(d) inoculating the second extract with lactic acid bacteria and incubating at 25 to 45° C. for 2 to 5 days to obtain a fermentation broth
Including,
The lactic acid bacteria in step (d) are Lactobacillus brevis DF01 or Pediococcus acidilactici K10.
The method of claim 5,
The step (b) is to extract hot water by mixing the pulverized heart and water in a ratio of 1:5 to 20 (w/v).
The method of claim 5,
In the step (c), the first extract is adjusted to pH 6 to 7 and then ultrasonicated.
delete The method of claim 5,
In the step (d), the initial inoculation concentration of lactic acid bacteria is 10 ⁷ to 10 ⁹ cfu/mL.
The method of claim 5,
After the step (d), the production method further comprising the step of filtering the fermentation broth of step (d).

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