KR101776143B1 - Pharmaceutical composition for preventing or treating schizophrenia comprising wood-cultivated ginseng extract - Google Patents
Pharmaceutical composition for preventing or treating schizophrenia comprising wood-cultivated ginseng extract Download PDFInfo
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- KR101776143B1 KR101776143B1 KR1020150181593A KR20150181593A KR101776143B1 KR 101776143 B1 KR101776143 B1 KR 101776143B1 KR 1020150181593 A KR1020150181593 A KR 1020150181593A KR 20150181593 A KR20150181593 A KR 20150181593A KR 101776143 B1 KR101776143 B1 KR 101776143B1
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- ginseng
- goat
- extract
- pharmaceutical composition
- schizophrenia
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Abstract
The present invention relates to a pharmaceutical composition for preventing or treating schizophrenia including an extract of goat ginseng. More particularly, the extract of goat ginseng, which is known to have a higher content of saponin and ginsenoside than that of cultivated ginseng, also has an action on the central nervous system And to a pharmaceutical composition for preventing or treating schizophrenia.
According to the present invention, based on the behavioral pharmacological evaluation including the positive symptoms, the negative symptoms and the cognitive memory impairment, and the biochemical evaluation as the antioxidant and oxidative stress indicator, There is an effect that can be exhibited as a composition for use.
Description
The present invention relates to a pharmaceutical composition comprising a goat's ginseng extract, and more particularly to a pharmaceutical composition for preventing or treating schizophrenia including a goat's ginseng extract.
Goat ginseng is rich in organic germanium and saponin, which have excellent pharmacological effects, and has health promotion effects regardless of age, including children and elderly people. Goat ginseng is produced in a mountain without artificial facilities, and is grown at a same position for more than 7 years. It is necessary to secure a cool mountain area with a summer temperature of 20 to 25 ° C, a lush shade with 80 to 90% of which is more than 20 years old, a weakly acidic sandy loam rich in organic matter and drainage, It is also tricky.
Ginseng is artificially managed by installing rain forest facilities in man-made fields, and it is divided into 4, 5, and 6 year old trees according to cultivation period. In order to increase the yield, it is thick by using appropriate fertilizer, compost, pesticide. On the other hand, goat ginseng can be cultivated for 7 to 10 years in a mountain. Wild or wild ginseng grown for more than 11 years is distinguished. The ginseng saponin and the biotissue constituents vary depending on the growth period of the ginseng, so that useful components are increased. However, since ginseng can not be cultivated for more than 7 years in field due to limitation of soil ability, it is known that ginseng is the most sensitive classification standard.
It is said that the goat ginseng grows in a shady, humid place such as a deep mountain birch, a lacquer tree, especially in a shade of about half, and it grows in a remote place, and it grows after 6 to 30 years after planting a ginseng seed in a mountain. In the case of ginseng, the head part (outsole) is about 3 ~ 7, whereas the goat ginseng is more than that according to the age. The roots of ginseng are thick and short but the goat ginseng is thin and long, sometimes exceeding 1 m. The life span of goat ginseng is 50 years ~ several hundred years depending on soil and climatic conditions, but ginseng is up to 20 years or so. It is characterized by strong. The efficacy of ginseng is enhanced by the stimulation and developmental effects on the central nervous system, activation of liver function, hypoglycemia, diabetes mellitus, inhibition of cancer cell growth, hypotension, prevention of arteriosclerosis, activation of immune system function, prevention of anemia, And stimulation of memory, stress and relieving fatigue.
In addition, the ratio of protopanaxadiol to protopanaxtriol was 3.25, which was higher than that of cultivated 4-year-old 2.45 and 6-year-old 2.18 in the comparative study of ginseng saponin of goat and cultivated ginseng In particular, the ratio of ginsenoside Rb1 to Rg1 was reported to be higher than that of goat ginseng at 10.22, 3.51 at 4 years and 4.86 at 6 years. In addition, the contents of Rd and Re have been reported to be higher in goat ginseng than in cultivated ginseng (Analysis of Ginsenoside Composition of Woods Grown Ginseng Roots, Sung Tai Han, Cha Gyun Shin, Byung Wook Yang, Young Tae Hahm, Byung Ok Im, Soon Hyun Cho, Boo Yong Lee and Sung Kwon Ko, Food Science & Biotechnology, 16 (2), 281-284, 2007).
In the current situation where the number of adult diseases such as cancer, heart disease and stroke is increasing along with the increase of the elderly population, people are increasingly interested in health. In order to prevent such chronic diseases, the third function of food, The development of food is becoming more emphasized.
In Korean Patent No. 10-0727435 (2007.06.05), which is a technique using pasty ginseng, there is no decay or deterioration even if it is left for a long time by removing water, and it can be easily ingested in water. However, Korean Patent No. 10-0799654 (2008.01.24) discloses a method for manufacturing a goat gum and a method for manufacturing a goat gum, which is useful for health, A method of manufacturing a soymilk for processing a very easy-to-eat form for the tasting of goat's ginseng, and a method for preparing the soymilk .
However, there is no report on the differentiated functionalities of goat ginseng, and there is no report on the central nervous system effect. Therefore, the present inventor completed the present invention in order to contribute to the increase in income of forestry workers by identifying the differentiation of the new functions through experiments related to the action of the central nervous system such as schizophrenia of the goat ginseng and developing the raw goat gins as processed products.
The object of the present invention is to provide a composition for preventing or treating schizophrenia by demonstrating the mitigating effect on the evaluation index of schizophrenia by comparing the efficacy of the goat's ginseng extract with that of existing cultivars.
In addition, the present invention aims at contributing to the increase of incomes of forestry workers by developing processed products including functional foods, etc., by confirming the differentiation of goat's ginseng against the existing cultivation ginseng.
In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating schizophrenia comprising extract of goat ginseng.
In one embodiment of the present invention said goat juice extract is selected from the group consisting of hallucinations, delusions, abnormal behavior, aphasia, autism, social isolation, spontaneous abortion, attention deficit and cognitive memory impairment The above symptoms can be alleviated.
In one embodiment of the present invention, the goat ginseng extract may be extracted from a sample harvested from the geochang-gun, Kyungsangnam-do.
According to the present invention, based on the behavioral pharmacological evaluation including the positive symptoms, the negative symptoms and the cognitive memory impairment, and the biochemical evaluation as the antioxidant and oxidative stress indicator, There is an effect that can be presented as a pharmaceutical composition.
Also, according to the present invention, it is possible to prove that the efficacy of the goat's ginseng extract is superior to the existing cultivated ginseng, and thus there is an effect of suggesting the possibility of producing a processed product such as a functional food.
FIG. 1 is a schematic diagram of a construction process of a schizophrenia-like mouse model according to an embodiment of the present invention.
FIG. 2 is a schematic diagram for performing (A) exercise activity test, (B) social interaction test, (C) forced swimming test, and (D) new object recognition experiment according to an embodiment of the present invention.
FIG. 3 is a graph showing the pharmacological efficacy of the extract of the goat ginseng against the positive symptoms in the schizophrenia-like mouse model according to the example of the present invention as data of the exercise activity experiment.
FIG. 4 is a graph showing the pharmacological efficacy evaluation of the extract of Aspergillus oryzae on negative symptoms in a schizophrenia-like mouse model according to an embodiment of the present invention as data of (A) social interaction and (B) forced swimming experiment.
FIG. 5 is a graph showing the pharmacological efficacy evaluation of the extract of the goat ginseng against the cognitive memory impairment in the schizophrenia-like mouse model according to the example of the present invention as data of a new object recognition experiment.
FIG. 6 is a graph showing the protective effect of the goat's ginseng extract on depletion of frontal glutathione concentration in a schizophrenia-like mouse model according to an embodiment of the present invention.
FIG. 7 is a graph showing the protective effect of the extract of the goat's ginseng on the oxidative stress index in the schizophrenia-like mouse model according to the present invention as (A) reactive oxygen species increase, (B) peroxide lipid increase, (C) protein carbonyl group increase Fig.
Hereinafter, the present invention will be described in detail.
The present invention provides a pharmaceutical composition for the prevention or treatment of schizophrenia including a goat's ginseng extract.
In one embodiment of the present invention, the goat extract is selected from the group consisting of hallucinations, delusions, abnormal behaviors, negative symptoms corresponding to positive symptoms which are evaluation index of schizophrenia, , Spontaneous abortion, attention deficit, and cognitive dysfunction. ≪ Desc /
In one embodiment of the present invention, the extract of the goat ginseng has an increase in glutathione concentration, increase of reactive oxygen species (ROS), increase of malondialdehyde (MDA) in the brain tissue which is an evaluation index of schizophrenia And an increase of the protein carbonyl group can be alleviated.
In one embodiment of the present invention, the goat ginseng extract may be extracted from a sample harvested from a geochang-gun, Kyungsangnam-do. The extraction may be carried out using water, a C 1 to C 4 alcohol, or a mixed solvent thereof. The extraction may be room temperature extraction, hot water extraction, cold extraction, reflux cooling extraction, ultrasonic extraction and steam extraction.
In one embodiment of the present invention, the pharmaceutical composition may further comprise at least one pharmaceutically acceptable carrier or additive. That is, the pharmaceutical composition of the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method . The pharmaceutically acceptable carrier may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. It also includes diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Solid form preparations for oral use include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like, and may include lubricants such as magnesium stearate and talc. Oral liquid preparations include suspensions, solutions, emulsions, syrups, and the like, and may contain diluents such as water and liquid paraffin, wetting agents, sweetening agents, fragrances, preservatives and the like. Examples of the non-aqueous solution include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations and suppositories. Non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, ethyl And injectable esters such as oleate. As a suppository base, witepsol, macrogol, tween, cacao butter, laurin, glycerogelatin and the like can be used.
The dose of the goat's ginseng extract contained in the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. For example, the goat's ginseng extract may be administered at a dose of 0.0001 to 1000 mg / kg per day, preferably 200 mg / kg per day, and the administration may be administered once a day or divided into several times. In addition, the pharmaceutical composition of the present invention may contain 0.001 to 90% by weight of the extract of Aspergillus oryzae in relation to the total weight of the composition.
Hereinafter, the present invention will be described in more detail with reference to examples and measurement examples. However, the following examples and measurement examples are provided for illustrating the present invention, and the scope of the present invention is not limited thereto.
Example 1. Preparation of goat ginseng extract
For the preparation of goat ginseng extract, the samples harvested from Gangchang - gun, Gyeongsangnam - do and Pyeongchang - gun, Gangwon - do were used respectively. Each sample was freeze-dried, and after centrifugation, 2,500 ml of distilled water was added to each 10 g of the sample, and the mixture was refluxed for 2 hours for 2 hours, filtered and concentrated under reduced pressure to prepare an extract. For use as a control, extracts of ginseng (6-year-old muscle) harvested from the negative under the same conditions as above were also prepared.
Example 2. Preparation of animal models
All animals were treated in accordance with the NIH Guide for Welfare and Use of Laboratory Animals (NIH Publication No. 85-23, 1985; www.dels.nas.edu/ila). The present invention has been carried out in accordance with ILAR (Institute for Laboratory Animal Research) guidelines for welfare and use of laboratory animals.
Male C57BL / 6J mice or male ICR mice (Bio Genomic Inc., Charles River Technology, Gapyeong-gun, Gyeonggi-do, Gyeonggi-do, Gyeonggi-do) were ad libitum maintained at 12 ± 12 h. Male ICR mice did not inject any drugs because they would be used as targets in the social interaction test.
10 mg / kg of PCP (phencyclidine hydrochloride, Tocris Bioscience, Ellisville, Mo., USA) was administered subcutaneously to male C57BL / 6J mice for 14 days to produce a schizophrenia-like model. The goat ginseng extract was administered by intraperitoneal injection at a dose of 200 mg / kg until the end of the behavioral
Example 3 Evaluation index for confirming the efficacy of goat ginseng extract
Example 3-1. Positive symptom evaluation
It is possible to evaluate positive symptoms by measuring locomotor activity using video tracking system (application of behavioral sensitization technique).
On the day before the experiment, mice were environmentally adapted more than once for 10 minutes in the test box. Place the mouse on the center of the test box and observe the behavior through the video tracking system for 10 to 30 minutes. Then, the total movement distance, the total movement duration, the total turn angle, Pattern and the like were analyzed (Fig. 2A).
Example 3-2. Negative symptom evaluation
It is possible to evaluate negative symptoms by performing social interaction tests and forced swimming experiments using a video tracking system.
For social interaction experiments, we set up an interaction zone in which a cage with a target mouse is placed inside a locomotor box. Experimental mice were placed in the corner zone and the time taken to travel to the interaction zone or the time spent in the interaction zone was monitored via a video tracking system (FIG. 2B).
For a forced swimming experiment to evaluate depressive symptoms, water was injected into a transparent cylinder of 20 cm in diameter and 40 cm in height to a depth of 20 cm, and then the mouse was moved for 4 minutes except for the initial one minute The immobility time was measured. Floating time was based on measuring the time when the mouse did not move all the limbs (Fig. 2C).
Example 3-3. Assessment of cognitive dysfunction
New object recognition experiments can be performed to assess cognitive memory impairment.
On the first day, mice were placed in a box of 40 cm x 40 cm x 30 cm and allowed to move freely for 10 minutes. On the second day, two objects were placed inside the box and the reaction time for each object was recorded. After 24 hours, one of the two objects was replaced with a novel object and the reaction time for the new object was recorded (Figure 2D).
Example 3-4. Assessment of antioxidant and oxidative stress indicators
For the measurement of oxidative stress markers and glutathione concentrations by biochemical evaluation, the frontal region of the mouse was sampled, ground with an ultrasonic grinding machine (Bronson Sonic, NY), and centrifuged at 10,000 g for 20 minutes at 4 ° C. The levels of glutathione and oxidative stress markers, reactive oxygen species (ROS), malondialdehyde (MDA), and protein carbonyl were measured.
To evaluate the extent to which ROS is formed, the conversion of 2 ', 7'-dichlorofluorescein diacetate (DCFH-DA) to dichlorofluorescein was performed according to a slightly modified method described by Bourre et al. Respectively. The frontal lobe homogenate was placed in a test tube containing 2 mL PBS and 10 nmol DCFH-DA and dissolved in methanol. The mixture was incubated at 37 [deg.] C for 3 hours and then absorbance was measured at 480 nm excitation wavelength and 525 nm emission wavelength. DCF was used as a reference material.
To determine the MDA content, quantitative evaluation was made by measuring the accumulation of thiobarbituric acid-reactive material in the frontal lobe homogenate (Behav. Brain Res., 155, 185-196). Briefly, 0.1 mL of homogenate (or a freshly prepared standard solution from 1,1,3,3-tetramethoxypropane) and 1 mL of a reaction reagent (0.37% thiobarbituric acid and 6.4% perchloric acid, 2: 1 v / v) 0.75 mL were mixed and heated to 95 DEG C in a water bath for 1 hour. After cooling (10 min in an ice bath), the precipitate was removed by centrifugation at 3200 x g for 10 min. The supernatant was neutralized, filtered and injected onto an
The extent of protein oxidation in frontal lobe homogenates was assessed by measuring the content of protein carbonyl groups. The content of the protein carbonyl group was measured spectrophotometrically using the 2,4-dinitrophenylhydrazine (DNPH) -labeling procedure (Behav.Brain Res., 155, 185-196). The results are expressed in nmol / mg of DNPH versus bound protein based on the extinction coefficient for an aliphatic hydrazone of 21 mM -1 cm -1 . Proteins were measured using a BCA protein assay reagent (Pierce, Rockford, Ill., USA).
Example 4. Statistical processing
Data were statistically processed using one-way ANOVA and Fisher's ratings were used for post-test. In each graph, each value represents the mean ± standard deviation of 10 individuals ( # P <0.01 vs. Saline + Saline, $ P <0.05, $$ P <0.01 vs. Saline + PCP).
Measurement example 1. Behavioral pharmacological evaluation of efficacy of goat ginseng extract
Administration of gecko ginseng extract to the positive symptoms of schizophrenia-like models induced in PCP administration as measured in Example 3-1 showed significant protective effects (FIG. 3).
Administration of the extract of goat ginseng showed a significant protective effect on the negative symptoms observed in the PCP-induced schizophrenia-like model measured in Example 3-2. In the social interaction experiments, the ginseng ginseng extract showed the most significant protective effect (Fig. 4A) and the ginseng extract was more effective than the negative ginseng extract in the forced swimming experiment (Fig. 4B).
In the case of cognitive memory impairment in the PCP-induced schizophrenia-like model as measured in Example 3-3, the administration of the extract of goat ginseng showed a significant protective effect (Fig. 5).
Measurement example 2. Biochemical evaluation of efficacy of goat ginseng extract
The administration of the extract of goat ginseng showed a significant protective effect on the reduction of the PCP-induced frontal glutathione concentration as measured in Example 3-4. The ginseng ginseng showed only a protective tendency, whereas the ginseng ginseng and pyeongchang ginseng showed a significant protective effect (FIG. 6).
The effects of PCP administration on the oxidative stress index showed significant protective effect. Significant protective effects of gaercombe ginseng were observed in both ROS, lipid peroxidation and protein carbonyl groups (Fig. 7).
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| Sung Wook Kang 외 4명. Panax ginseng attenuates MK-801-induced stereotypy in mice. 스트레스연구. 제17권 제4호, 2009년 12월, pp.435-440* |
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