KR20180070403A - Mixed Naturopathic Compositions of Zizyphus jujuba Miller and Magnolia officinalis Containing Anti-depressant Components by Supercritical Fluid Extraction Process - Google Patents

Abstract

The present invention relates to a mixed extract of Zizyphus vulgaris and Machilus thunbergii extracted by using a supercritical fluid extraction process, and to a pharmacological composition and health food containing the same. More specifically, the mixed extract of Zizyphus vulgaris and Machilus thunbergii exhibits antidepressant activity through an animal behavioral method and safety is secured through a cytotoxicity test, so that the mixed extract of Zizyphus vulgaris and Machilus thunbergii can be usefully used as a composition for treating depression and health food.

Description

[0001] The present invention relates to a natural pharmaceutical composition comprising an acid depressant component and an acidophilus component, which comprises an antidepressant component by a supercritical fluid extraction process, and a supercritical fluid extraction process,

The present invention relates to the use of a mixed extract of a sanjoin and a japonicum as a pharmaceutical composition through a supercritical fluid extraction process, and more particularly to a pharmaceutical composition comprising an antidepressant effect using a natural acidophilus and a mixture of a natural acidophilus and a horsetail, Or memory loss and a health functional food.

     From ancient times to modern times, the reality of melancholia has been recognized to include symptoms of both depression and anxiety (Glass GA, 1994. A conceptual history of anxiety and depression. JA, Sitsen JMA, Handbook of depression and anxiety, New York, NY: Marcel Dekker, p. Anxiety or depression are very common with each other. 10-25% of the total population will experience some depression at some point in their lives, and 5-6% will be suffering right now. Depression can range from young students to the elderly, and women appear earlier, but most are in their late 30s and early 40s. Depression usually lasts for six months or longer, suffers from physical and mental suffering, and may suicide without appropriate treatment. Anxiety is expressed by emotional symptoms, physical symptoms, thought symptoms, emotional symptoms are restlessness, irritability and sensitivity. Physical symptoms include heart leaps, digestive problems, diarrhea, constipation, There are sweating on the hands, trembling hands and body, hands and feet, muscle tension, headache, nape or shoulder pulling pain, pressure or pain in the chest, dry mouth, shortness of breath, dizziness and insomnia.

     Anxiety and depression are also known to be closely related to memory impairment. There is evidence that depression or anxiety can occur as a result of memory decline [Schmand, B., Jonker, C., Geerlings, M. I., Lindeboom, J. Subjective memory complaints in the elderly, depressive symptoms and future dementia. Br. J. Psychiatry 171, 373-376,1997; Harwood, D. G., Barker, W. W., Ownby, R. L., Duara, R. Relationship of behavioral and psychological symptoms to cognitive impairment and functional status in Alzheimer's disease. Int. J. Geriatr. Psychiatry 15, 393-400, 2000; Clarnette, R. M., Almeida, O. P., Fors, H., Paton, A., Martins, R. N. Clinical characteristics of individuals with subjective memory loss in Western Australia: results from a cross-sectional study. Int. J. Geriatr. Psychiatry 16, 168-174, 2001].

     It has also been reported that anxiety and depression are caused not only as a cause of memory loss but also as a response to the onset of memory decline (Devanand, D. P., Sano, M., Tang, M. X. et al. Jenn, AF, Christensen, H., Korten, AE, Hendersen, < RTI ID = 0.0 > In this study, we examined the relationship between memory impairment, anxiety, and depressive symptoms in children with memory impairment and anxiety. Is known to be bi-directional because anxiety leads to amnesia [Derouesne, C., Lacomblez, L., Thibault, S., LePoncin, M. Memory complaints in young and elderly Psychiatry 14, 291-301, 1999], amnesia also causes anxiety [Schneider, LS Overview of generalized anxiety disorder in the elderly, J. Clin. Psychiatry 57, 45, 1996]. Thus, the importance of these two- Billion loss is the fact that you can not share a close relationship with each other [Sinoff, G., Werner, P. Anxiety dosorder and accompanying subjective memory loss in the elderly as a predictor of future cognitive decline. Int. J. Geriatr. Pychiatry 18, 951-959, 2003]. Anxiety and depression are common in 25-50% of patients, and these comorbidities increase the severity of anxiety or depression [Kessler RC et al., (1999). Lifetime comorbidities between social phobia and mood disorders in the US national comorbidity survey. Psychol med 29, 555-567].

     Currently, benzodiazepines based on GABA receptors are used as antihypertensive drugs, but they have side effects such as premature forgetfulness, movement disorders, mental disorders, and confusion; Currently, acetylcholinesterase inhibitors are used for the treatment of dementia. However, they are generally not effective. Most commonly used tacrine is used in one third of the patients with abdominal cramps, appetite deficiency, nausea, vomiting and diarrhea And other acetylcholinesterase inhibitors such as donepezil, ribastigmain, and galactamine, which are also restricted in their use due to side effects such as nausea, vomiting, diarrhea and insomnia [Harman JG, Limbird, LE Goodman & Gilman ' s The Pharmacological Basis of Therapeutics. 10th edition, McGraw Hill, 2001].

     As antidepressants, tricyclic antidepressants, monoamine oxidase inhibitors (MAOIs) and selective serotonin reuptake inhibitors (SSRIs), which suppress the reabsorption of monoamines into the nerve endings, have been used. Tricyclic antidepressants can cause insomnia, anxiety, fatigue, weakness, dry mouth, and pupil dilation due to harmful effects. In patients with heart disease, sudden death may occur in ventricular arrhythmias or myocardial infarction even at the normal dose. In the case of MAOI, it causes side effects such as hepatotoxicity and somatic hypotension, and overdose causes insomnia, excitement, and convulsions. Significant side effects of SSRIs lead to gastrointestinal disorders such as diarrhea, nausea and vomiting, and cause anxiety, anxiety, sleep disorders, weight gain, sexual dysfunction and withdrawal [Harman J. G., Limbird, L. E. Goodman & Gilman ' s Pharmacological Basis of Therapeutics. 10th edition, McGraw Hill, 2001]. Therefore, it is required to develop a substance which can treat anxiety, depression and memory disorder simultaneously as a drug with less side effects and excellent effects.

     In general, an organic solvent such as methanol, ethanol, hexane or the like is used as a typical method for extracting an active ingredient from plants and plants. The extraction method using an organic solvent is easy to apply, while selection of an appropriate solvent, Difficulty in solvent removal, environmental pollution, low extraction yield, and usually requires a long extraction time of about 24 to 48 hours. However, for commercial use, it is necessary to shorten the stability to the solvent and the extraction time, to acquire environmental pollution and the ingredients to be extracted to the maximum extent, and to have activity, but there is no clear method for this.

     Supercritical Fluid Extraction Technology is a technique using fluid above critical temperature and critical pressure. Critical temperature and pressure are the temperature and pressure point at which the phase transition from the gas phase to the liquid phase occurs. It changes the temperature pressure volume, Refers to the temperature and pressure in a state in which no change takes place at any point when a change such as liquefaction, vaporization of liquid, or the like occurs. For example, ammonia is known to be at 132 ° C, carbon dioxide at 31 ° C, and oxygen at -119 ° C, which is known as the critical temperature. At higher temperatures, no matter how large the pressure is, the gas does not liquefy.

     Among the candidates for supercritical fluids, carbon dioxide is not only present in an infinite amount in nature, but also in the steel industry or petrochemical industry. It is colorless, odorless, harmless to the human body and chemically very stable. Above all, it has a lower critical temperature (31.1 ℃) and a critical pressure (7.4 MPa) than any other fluid and can be easily adjusted to supercritical conditions, resulting in higher energy efficiency. In addition to this, it is possible to solve the problems occurring in the existing processes, that is, the human toxicity due to the organic solvent remaining in the final product.

     Supercritical extraction has attracted attention as a new environmentally friendly cleaner technology that can replace existing processes in the field of extraction, purification, etc. of pharmaceuticals, food processing, petrochemical refining, etc. (CD Bevan and PS Marshall. Nat. Prod., Rep. 11, 451-576, 1994; WK Modey, DA Mullholl et al., Phytochem.

     The fields where the early supercritical fluid technology is mainly applied to the extraction and purification of natural materials are limited to nonpolar materials such as spices, cosmetics, and fats, and low-cost foods and fragrance components. Recently, the development of various phenomenological characteristics and additional technologies Polar, small amount, and expensive natural drug extraction and purification.

     On the other hand, Sanjin (Zizyphus jujuba Miller) is a kind of jujube, a broad-leaved arboreous tree belonging to the Fhamnaceae family. It is widely grown from Europe to Asia. In one room, there are three kinds of plants, such as control (大 夏), dried (夏 夏), swan (白 棗), grape (桂 夏), brew (棗), 조 (棗) It is also called hydration, and it can be used as a sedative, hypnotic action, heart failure, stomach cramps, circulatory diseases, hepatitis, cirrhosis, diuretic, tonic, emollient, anemia, anorexia, nervous breakdown and hysterical treatment, , Bronchitis, tuberculosis, hemorrhagic disease, and expectorant drugs.

     It is described in Korean Patent No. 10-0931880 that a herbal medicine containing a sanjoin, an oriental soya, a soya sauce and a raw paper has a depression relieving effect and a heart attack prevention effect by solving insomnia Korean Patent No. 10-0763620 discloses that a sanjoin extract is used as a composition for promoting growth hormone secretion and Korean Patent No. 10-1071154 discloses an anti-stress composition using a sedative effect of a sanjoin have.

     In addition, the sanjoin is used as an effective ingredient of the external preparation for skin, Korean Patent Laid-Open No. 10-2005-0026806 discloses that the acidophilus exhibits an effect of prolonging the sleeping time and suppressing the secretion of cortisol by the stress hormone, thereby being used as a cosmetic composition .

     Magnolia officinalis is an evergreen broad-leaved tree belonging to the dicotyledonous plant, and is an evergreen broad-leaved tree belonging to the family Ranunculaceae. It lives in Korea, Japan, Taiwan, and southern China. As a medicinal herb, stem and egg shells have been used in the treatment of asthma and gastrointestinal disorders and contain ingredients such as magnolol, isomagnolol, honokiol, and machiol. Among them, honokiol belongs to polyphenol and is used as a useful substance showing antidepressant, thrombolytic inhibition, nerve stabilization and antibacterial activity. Recently, it has been reported that honokiol induces cytotoxicity through induction of mitochondrial permeability transition pore and activation of cytochrome crease and caspase of various cancer cells. (JY Yang, MA DellaFera, S. Rayalam and CA Baile, Enhanced effects of xanthohumol plus honokiol on apoptosis in 3T3-L1 adipocytes. Obesity, 16 (6), 1232 (2008)).

     The results of this study suggest that the antidepressant effect of Sanjo and Yakju is likely to be antidepressive. However, only medicines related to antidepressant medicines such as ginseng, jujube, livestock, There is no study on the action of the acidophilus.

Accordingly, the present inventors have conducted studies on substances which cause prevention, treatment and memory enhancement of anxiety, depression and dementia of modern people who are suffering from brain damage due to various environmental factors such as stress, drinking, smoking, The present invention has been accomplished based on this discovery that it exhibits excellent effects in preventing and treating anxiety, depression and dementia, and enhancing memory.

KR 10-2006-0041105 KR 10-2013-0060835 KR 10-2006-0083267

It is an object of the present invention to provide a pharmaceutical composition exhibiting an effect of inhibiting anxiety, depression, memory and dementia by using a mixture of Sanjoin and Lilium japonica extract. Especially, it is a main object to effectively prevent and treat the mutual anxiety, depression and memory disorder which interact with each other by using a combination extract of Sanjoin and Zucchini having both an anticancer and antidepressant and memory enhancing effects.

In the present invention, the extracts maximizing the antidepressant effect using the supercritical fluid extraction technique in the acidophilus and ascidian, which are natural substances estimated to have an antidepressant effect, Provide medicines.

As described above, the supercritical fluid extraction method can extract the antioxidant substances derived from the acidophilus such as acidophilus and acanthin in a short extraction time without residual organic solvent, and can relieve anxiety, improve seizure, sedation, It is possible to extract anti-depressant substances that are effective for improvement, and it can be applied to industrial fields such as foods and medicines.

FIG. 1 is a process diagram of a supercritical fluid extraction process of the present invention using a supercritical fluid extraction process.
2 is a graph showing the immobility time in the FST
3 is a graph showing the swimming time in the FST.
4 is a graph showing the climing time in the FST.
5 is a graph showing immobility time in TST.
6 is a graph showing the survival rate in the MTT asay experiment.

     Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention and are not intended to limit the present invention.

<Example 1> Production of antioxidant acidophilus and ascorbic acid complex extract

<1-1> Drying and crushing stage of natural acidophilus

     Prior to the extraction of Sanjo (Zizyphus jujuba Mill var. Spinosa Hu) and Magnolia officinalis L., steps were taken to dismantle. At this time, the samples were frozen at 0 ~ 10 ℃ using a freeze dryer and then dried at 0.001 ~ 0.006atm. Dried sanjoin and latex were crushed using a mixer.

<1-2> Hot water extraction method

     The acidophilus and hinoki were dried, ground finely with a grinder, and heated in a hot water bath to obtain a mixture of acidophilus and hinoki extract. At this time, about 1 L of distilled water was added to 50 g of each of acidophilus and ascidian samples, and the mixture was heated for 3 hours in a hot water of 100 ° C. and extracted. The extract was kept at a constant temperature (90 ° C.) The filtrate was concentrated in a rotary vacuum concentrator.

&Lt; 1-3 > Organic solvent extraction method

     In order to prepare the solvent extract, the acidophilus and the ascorbic acid were dried and finely pulverized by a pulverizer, and then immersed in ethanol for 24 hours. After centrifugation, only supernatant was separated and filtered under reduced pressure with filter paper. The organic solvent extract was concentrated under reduced pressure with a rotary vacuum concentrator at 40 ° C to prepare a sample.

<1-4> Preparation of Supercritical Fluid Extract of Acanthopanax acantholum

     The acidophilus and hindsack were dried and 2 kg of the sample crushed at a ratio of 1: 1 and 1 L of the co-solvent ethanol were added to the supercritical fluid extraction reactor, and the reactor was extracted under the conditions that the temperature was 65 ° C and the pressure was 350 bar. For the supercritical fluid extraction in the following example, 65 ° C and 350 bar were the best results, and this condition was used in the present invention. The extract was dried with a rotary evaporator to obtain a mixture of the acidophilus and succulent supercritical fluid. High-performance liquid chromatography (HPLC) was performed under the following conditions in order to analyze supernatant fluid extracts of the obtained acidophilus and ascidian. These results show that the supernatant fluid extraction method can effectively extract the essential oil components from the acidophilus. Therefore, we tried to establish the optimal condition for extracting essential oil components from Sanjia and Yakju.

&Lt; Preparation Example 1 > Optimization of supercritical fluid extraction time of acidophilus and hippurate

     In Preparation Example 1, supercritical fluid extraction parameters other than the extraction time were fixed, and the extraction time was changed by changing the extraction time to establish the optimum extraction time. Using the supercritical fluid extraction device used in Example 1-4, 0.5 g of the acidophilus powder was extracted, the extraction temperature was 65 ° C., the extraction pressure was 350 bar, the supercritical fluid carbon dioxide flow rate was 5 g / min And the time was changed from 2 hours to 3 hours and 30 minutes at intervals of 30 minutes. The results are shown in Table 1 below.

Extraction amount of essential oil component [mg / g] 2 hours 2 hours 30 minutes 3 hours 3 hours and 30 minutes 65 ℃
350 bar 12.94 15.28 19.63 17.70

     According to the above Table 1, the extraction amount of the essential oil component gradually increased as the extraction time increased to 2 hours, 2 hours 30 minutes, and 3 hours, but the extraction amount of the essential oil component decreased rather than 3 hours 30 minutes I could. Therefore, extraction of essential oil components from acidophilus and ascidian using supercritical fluid carbon dioxide showed the best extraction yield when extracted for 3 hours.

PREPARATION EXAMPLE 2 Optimization of Supercritical Fluid Extraction Pressure and Temperature of Acidophil

     In Production Example 2, the optimum extraction temperature and pressure were established by confirming the extraction amount while changing the temperature and pressure at the optimal extraction time (3 hours) obtained through Production Example 1 above. Supercritical fluid Extraction pressure of carbon dioxide was changed from 350 bar to 500 bar at 50 bar intervals and supercritical fluid extraction was performed. The temperature was changed from 65 ℃ to 75 ℃ at 5 ℃ intervals to extract the supercritical fluid. The results are shown in Table 2 below.

Extraction amount of essential oil component [mg / g] 65 ℃ 70 ℃ 75 ℃ 350bar 19.63 11.95 9.67 400bar 17.12 16.91 12.18 450bar 10.55 14.15 12.10 500bar 7.98 13.86 11.95

According to the above table 2, when the extraction pressure is 350 bar, the extraction amount decreases as the temperature increases. When the extraction pressure is 350 to 500 bar, the extraction amount increases and then decreases again as the temperature increases. Also, when the temperature was 65 ℃, the extraction amount decreased with increasing pressure. When the temperature was 70 ~ 75 ℃, the extraction amount increased and then decreased again with increasing pressure. In addition, extraction volume was the highest at extraction pressure of 350 bar and temperature of 65 ℃.

&Lt; Example 2 > Test for anti-depressive effect of a mixture of acidophilus and rosemary extract

     The anti-depressant effect was confirmed by using the mixed-type supercritical fluid extract (hereafter abbreviated as 'mixed supercritical extract') of the acidophilus prepared from Example 1-4.

<2-1> Design of experiments

Animal preparation

     The animals used in the experiment were male Sprague-Dawley rats weighing 240-270 g and purchased from Samtaco Animal Co. (Seoul, Korea). Rats were kept in restricted access facilities for up to 5 animals per polycarbonate cage. The cage temperature was maintained at 22 ° C ± 2 ° C and the relative humidity was maintained at 55-60%. The cage was ignited by artificial light for 12 hours daily. Sterile drinking water and a standard feeding diet were randomly fed into each cage during the experiment. The breeding indoor ventilator and the air purifier were always operated. Animal testing was conducted in accordance with the National Institutes Health Guide to Laboratory Animals Management and Use (NIH Publication No. 80-23), revised in 1996. All animals were used for one week after arrival at the feeding room.

Classification of experiment group

     The animals used in the experiment were randomly divided into 4 groups as follows.

1) Normal group: Normal group (5 animals) without administration of stress (immobilization stress)

2) Control group: Groups (5 animals) were administered with 1 ml / kg of physiological saline,

3) Low concentration group: A group of stress (5 rats) orally administered with 200 mg / kg of mixed supercritical extract of Sanjoin &

4) High concentration group: A group of stress (5 rats) orally administered at 400 mg /

Statistical processing

     All measurements were expressed as means ± standard error (mean ± SEM), and statistical analysis was performed by one-way ANOVA of the SPSS program for windows (ver. 11.0). Statistical significance was significant at p <0.05.

<2-2> Forced swimming test (FST)

     The antidepressant activity by forced swimming (FST) was measured by Porsolt et al. The first FST was performed after having the adaptation period of the incubation room for one week. Five mice per group were forced into a cylinder (25 cm in diameter) filled with water of 23 ± 1 ° C at a level of 40 cm, at which the rat's tail did not reach the bottom, and swim for 15 minutes. The time it took to reach immobility for the first time while observing the swimmers was recorded. Twenty-four hours later, the second FST experiment was performed for 5 minutes. The low-concentration group (200 mg / kg) or the high-concentration group (400 mg / kg orally) was administered 30 minutes before the experiment. For five days, mixed supercritical extracts were orally injected once a day, and the last FST experiment was run for 5 minutes. At this time, rats were measured for climbing, swimming, or immobility, respectively. Typical immobility is that the mouse is only partially exposed to the surface of the body, including the face, while maintaining body balance and no movement of the limbs. Swimming is a state in which a rat moves around the surface of the water and sometimes dives under water. Climbing is the most intense exercise, and you are going to climb above the acrylic cylinder by actively using your forelegs. To reduce errors and errors caused by human errors, we measured the data with a video camera.

<2-3> Forced swimming test (FST) results

     As a result, as shown in Table 3 and FIG. 2, in the immobility test, the immobility time was reduced by 50% or more in the mixed supercritical extract administration group compared with the control group administered with the physiological saline solution, and the mixed supercritical extract low concentration 200 mg / kg orally) and high concentration (400 mg / kg orally) administration group. In addition, we measured not only the immobility time but also the aggressive climbing and swimming behavior. As shown in FIG. 3 and FIG. 4, in the above experiment, the swimming time was decreased and the aggressive climbing time was increased in the mixed supercritical extract group than the control group.

FST Control group Low dose group (200mg / kg dose) High dose group (400mg / kg dose) Average Standard error Average Standard error Average Standard error Immobility 19.10 7.21 6.67 2.96 6.91 4.92 Climbing 51.17 7.54 135.21 14.55 76.97 12.95 Swimming 238.25 6.86 193.28 17.01 198.95 15.85

 (Unit: sec, sec)

<2-4> Tail suspension test (TST)

     Measurement of antidepressant activity by the tail suspension method (TST) was performed by Steru et al. Tail-dangled mice exhibit activity or immobility, using tape to hang the mice upside down. Under the red light, the patient is suspended for 6 minutes in a state where the noise is blocked, and the depression behavior is evaluated by measuring the time taken for floating action for 4 minutes without measuring for 2 minutes.

<2-5> Tail suspension test (TST) results

     As a result, as shown in Table 4 and FIG. 5, the control group remained immobilized for a remarkably long period of time, compared with the normal group, and the supercritical extract of low concentration (200 mg / kg orally) and high concentration (400 mg / The duration of immobility was shorter in the treated group than in the untreated group and the duration of immobility was shorter in the high dose group.

TST Control group Low dose group (200mg / kg dose) High dose group (400mg / kg dose) Average Standard error Average Standard error Average Standard error Immobility 122.95 11.5 124.13 10.8 72.15 6.95

 (Unit: sec, sec)

<Example 3> Stability test of a mixture of acidophilus and succulent supercritical extracts using MTT assay

     It is necessary to confirm the stability of the sanjoin and sukbai mixed supercritical extract prepared in Example 1-4 for use as a drug. Thus, MTT assay experiments were conducted to test whether the cells were toxic for use as a drug.

<3-1> Mixed supernatant extract of MTT Assay

     Neuronal cell line SH SY-5Y cells were plated at a density of cells / well in a 12-well plate and cultured at 37 ° C in a 5% CO 2 incubator. Each sample was treated for each concentration and cultured for 24 hours. After culturing, 100 μL of MTT was added to each well and incubated for 4 hours. Then, the supernatant was removed and formazan formed was dissolved in 100 μL of DMSO (dimethylsulfoxide) solution. After that, the absorbance was measured at 570 nm using a microplate reader, and the absorbance difference between the addition of the sample solution and the no addition was expressed as a percentage.

<3-2> MTT assay result of supernatant extract of mixture of acidophilus and glutinous rice

     As a result, as shown in FIG. 6, cell survival was decreased at a concentration of 0.3 to 0.4 mg / well. If interpreted differently, there is no big problem for cells to survive up to a concentration of 0.3 mg.

&Lt; Example 4 > Absorption degree of extract components for maintenance selection of soft capsules

      The absorbency of the active ingredients of the soft capsule of the present invention was measured for each suspension, and the results are shown in Table 3 below. The absorbency of the fat is represented by the number of gauge of the fat base required to suspend 1 g of the active ingredient into a moldable suspension. For the maintenance agent, 1 ~ 5% lecithin was used as a wetting agent. All experimental values in Table 5 below are in units of g.

Sanjo and Yakju
Mixed Supercritical Extract Time and peppermint
Mixed Supercritical Extract MCT oil 0.3 0.5 Soybean oil 0.5 0.7 olive 0.5 0.7 Grape seed oil 0.5 0.8 Evening Primrose Oil 0.6 0.8 Small-leaved ayu 0.5 0.6 Refined fish oil 0.6 0.7

 (Unit: g)

      From Table 5, it can be seen that the MCT oil, which is a kind of refined processed oil, has the best absorption characteristics. Accordingly, the present inventors have selected the MCT channel as the holding of the soft capsule.

<Example 5> Preparation of soft capsules

200 mg of a mixed extract of Sanjoin and hagi were mixed with 250 mg of MCT oil, 34 mg of palm oil and 16 mg of white wax to prepare 500 mg of the active ingredient mixture. Further, 1 part by weight of mint flavor essence was added to a capsule base containing 70 parts by weight of gelatin, 20 parts by weight of glycerin and 10 parts by weight of sorbitol solution per 100 parts by weight of the capsule base, and the mixture was mixed with ethyl vanadate Capsule coatings were prepared by appropriately adding tar pigments and red iron oxide. The capsule of the active ingredient was packed into a capsule by a molding machine, and the capsule was formulated into capsules by pressure and heat. The capsule was dried and coated to prepare the epilatorized soft capsule according to the present invention.

     Using the extraction method presented in the present invention, it is possible to produce the acidophilus extract and the extract of Zucchini, which have more antidepressant properties than the extracts extracted by the conventional extraction method.

Claims (3)

1) Processing step of crushing acidophilus,
2) filling the supernatant fluid extractor with the acidophilus powder crushed in step 1
3) adding a supercritical fluid to the extractor filled with powder through a heat exchanger in step 2, and then extracting the extract
4) Preparation method of extracts containing high content of antidepressant substance by supercritical fluid extract isolated in step 3 1)
2) filling the supernatant fluid extraction device with the acidophilus powder crushed in step 1;
3) Extracting the powder filled in step 2 for 3 hours at 350 bar and 65 ° C using a supercritical carbon dioxide extracting apparatus to prepare the acidophilus and supercritical extract
4) Preparation of high-content extract of antidepressant substance by supercritical fluid extract isolated in step 3       The pharmaceutical composition for prevention and improvement of depression according to claim 1, wherein the weight ratio of the extract to the extract (1) is in the weight ratio (w / w) The soft capsule preparation according to claim 1, wherein the MCT oil according to the fifth embodiment of the present invention is used as a prescription method.

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