KR20170141845A - A Pharmaceutical Composition comprising Crataegus pinnatifida extract for prevention or treatment of depressive disorders - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising a Crataegus pinnatifida extract, as an active ingredient, for prevention or treatment of depressive disorders. The composition of the present invention is effective in reporting nerve cells and treating or ameliorating depressive disorders. Therefore, there are advantages of overcoming the side effects of conventional antidepressants and at the same time bringing about an effect of treating the depressive disorders.

Description

[0001] The present invention relates to a pharmaceutical composition for the treatment or prevention of depression comprising Crassus extracts,

The present invention relates to a therapeutic or prophylactic pharmaceutical composition for depression comprising Crassulaceae extract.

Depression is an emotional pathology that occurs regardless of the objective situation. All lives of the patient are covered with depressed moods, decreased in interest, become anhedonia, become depressed, depressed, and desperate. It is a disease that reaches to the suicide prayer because it is motivated and it shows various physical symptoms such as loss of appetite, insomnia, constipation, loss of libido. Such depression has become a serious social problem in recent years.

Current antidepressants have a pharmacological action that increases the concentration of neurotransmitters in central serotonin or noradrenaline synapses. Antidepressants can be classified as tricyclic antidepressants (TCA), monoamine oxidase inhibitors (MAOI), or selective serotonin reuptake inhibitors (SSRIs) depending on the mechanisms that increase the concentration of neurotransmitters. And so on.

Although these existing antidepressants are relatively effective, some of them may have serious side effects and effects that do not meet expectations. (Geminick and Nemeroff, 2000). Inadequate response to antidepressant drugs (Thase et al., 2001), high recurrence rates (Frank et al. Tricyclic antidepressants (TCA) imipramine, desipramine, etc., have severe side effects such as hypotension and cardiac dysfunction. The most commonly used selective serotonin reuptake inhibitors, such as fluoxetine and sertraline, cause zone, gastrointestinal bleeding or sexual dysfunction.

Crataegus pinnatifida ) belongs to Rosaceae ( Crataegus) pinnatifida Bunge ) is a mature fault, also called mountain hunting or mountain hunter. Dried sea bream has been used as a digestive aid. (Korean Patent No. 10-0570120), the effect of treating degenerative brain diseases such as Alzheimer's (Korean Patent No. 10-1194974), the effect of treating prostate-related diseases 10-1607507), subclinical effects (Korean Patent No. 10-0965623), and hair loss prevention effect (Korean Patent Laid-Open No. 10-2010-0007003), but effects related to depression have been reported none. Accordingly, the present inventors intend to provide a pharmaceutical composition containing the horseshoe extract effective for the treatment of depression without the side effects described above.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have tried to develop a therapeutic agent for depression using natural extracts in order to improve the side effects of conventional antidepressants. As a result, the present inventors have completed the present invention by confirming that the Crassulaceae extract is effective for the treatment or improvement of depression.

It is therefore an object of the present invention to provide a therapeutic or prophylactic pharmaceutical composition for depression.

It is another object of the present invention to provide a composition for improving or preventing depression.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the invention there is provided hawthorn (Crataegus pinnatifida ) extract as an active ingredient.

According to another aspect of the invention there is provided hawthorn (Crataegus pinnatifida ) extract. < / RTI >

The inventors of the present invention have made efforts to develop a therapeutic agent for depression using natural extracts in order to improve adverse effects in existing antidepressants, and as a result, it has been confirmed that the extract is effective for the treatment or amelioration of depression.

The term " extract " used herein to refer to the hornblende extract includes not only the extraction result obtained by treating the hornblende with the extraction solvent but also the hornblende processed into a formulated (e.g., powdered) It has meaning.

The horseshoe crab extract used in the present invention can be obtained either by purchase or by direct extraction. In the case where the horseshoe extract used in the composition of the present invention is obtained by treating an extractant in an acid solution, various extraction solvents can be used. Preferably, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF (Tetrahydrofuran).

Alternatively, the horseshoe extract used in the present invention can be extracted with water, a C ₁ to C ₄ lower alcohol, or a mixed solvent thereof. According to the present invention, the composition of the present invention may comprise an extract of Sasa sp. Containing ethyl alcohol as a solvent. An example of a method for directly extracting the horsetail extract is as follows. 600 g of sorghum was added to 6 L of a 10-fold aliquot, and the mixture was extracted with a vacuum low-temperature extraction apparatus (COSMOS-660) at 60 ° C for 6 hours. The extracted concentrate was lyophilized to obtain an acid-extracted powder. In the present invention, crude crustacea extracted by a solvent can be used as the Crassulaceae extract and can be purified and used in high purity.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the crustacea extract includes not only those obtained using the above-mentioned extraction solvent, but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained by the purification method is also included in the crustacea extract of the present invention.

The crustacea extract used in the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray drying.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve the following efficacy or activity. The present invention is a composition extracted from a natural plant material, which is a natural plant material. Since the composition does not adversely affect the human body even when it is administered in an excessive amount, the quantitative upper limit of the composition of the present invention can be selected by those skilled in the art.

As demonstrated in the following examples, the administration of Crassostrea japonica extract showed nerve cell protective effect and showed an anti-depressive effect due to an increase in swimming time in a forced swimming experiment. Such experimental data show that the composition of the present invention can be used for the prevention or treatment of depression and can be used as a substitute for conventional antidepressants.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Such pharmaceutically acceptable carriers are those conventionally used in the field of manufacture and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, But are not limited to, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10000 mg / kg.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

When the composition of the present invention is a food composition, it may be prepared in the form of powder, granule, tablet, capsule or beverage. For example, various foods of candy, beverages, gums, tea, vitamin complex, or health supplement foods.

The food composition of the present invention may contain, as an active ingredient, not only horseradish extract but also components that are ordinarily added in food production, including, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, mulberry extract, jujube extract, licorice extract, have.

The features and advantages of the present invention are summarized as follows:

(a) Crataegus pinnatifida ) extract as an active ingredient.

(b) The composition of the present invention is effective in reporting nerve cells and treating or ameliorating depression.

(c) This has the advantage of overcoming the adverse effects of the antidepressants of the prior art, and at the same time being capable of treating depression.

FIG. 1 is a graph showing the protective effect of the extract of Sansa sp. In the dexamethasone-supplemented cell line (DEX), the survival rate was about 68% depending on the cytotoxicity, but the survival rate was up to 124% (100 μM) in the cell line supplemented with the sansa mushroom extract and the concentration of 50 μM and 100 μM The antidepressants imipramine and the existing antidepressants. Cell survival rate was significantly higher than that of the cell line treated with John's wort. The control group was treated with 1% DMSO (dimethyl sulfoxide) solvent.
FIGS. 2A and 2B are graphs showing results of forced swimming experiments after injecting the extract of Sansa sp. The group of oral administration of Sansa sp. Extract significantly decreased the immobility time (Fig. 2b) compared to the group (Con) of the second distilled water administered orally, and the swimming time increased (Fig. 2A).

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Experimental Methods and Materials

Sansa extract

The Crataegus used in the experiment pinnatifida ) was purchased from Kangwon, and 600 g of Sansa was added to 6 L of 10-fold alcohol, and then extracted at 60 ° C for 6 hours using a vacuum low-temperature extraction apparatus (COSMOS-660). The extraction concentrate was lyophilized to obtain an acid - extracted powder, and the extraction yield was 23.1%.

Cell culture

SH-SY5Y, a human neuroblastoma cell line, was distributed to ATCC (Manassas, Va., USA), and 10% FBS (Gibco, Carlsbad, Calif., USA) USA) and 1% penicillin streptomycin (gibco, USA) supplemented with Dulbecco's modified Eagle medium (Gibco) medium at 37 ° C and 5% CO ₂ , and subcultured at 2-3 days intervals .

Depression is associated with abnormal activity of the hypothalamus-pituitary-adrenal axis (HPA axis), and thus hyperglial extracellular glucocorticoid acts as a neurotoxic substance, resulting in atrophy and cell death of the brain nerve cells. Thus, SH-SY5Y, a cell line similar to human brain cells, causes cytotoxicity due to dexamethasone, a synthetic glucocorticoid-based component, to cause brain cell death by a mechanism similar to that of depression.

Animal experiment

Male ICR mice weighing 25g to 30g (5 weeks old, Koatech, Gyeonggi-do, Korea) were kept at a control temperature of 23 ± 1 ℃ for 5 animals per cage to evaluate antidepressant efficacy through animal behavior test. A dark cycle was performed. The mice were adapted for one week. The animal protocol was approved by the Korea Food Research Institute's Institutional Animal Care and Use Committee (IACUC). In order to measure the swimming time and swimming time according to the forced swimming behavior test, mice were orally administered a sample of 300 mg / kg (0.1 mL / 10 g _weight ) dissolved in the second distilled water, Experiments were performed.

The forced swimming behavior test is the most commonly used depressive animal model, in which a transparent acrylic cylinder (20 cm in height, 12 cm in diameter) is filled with water at 22-24 ° C at a height of 10 cm and forced swimming of the mouse. In this case, the immobility of the mouse is the behavior that occurs when you give up in an environment that can not escape from the water. As the immobility time increases, the state of the mouse becomes physiologically very similar to that of human depression. The existing antidepressants of various pharmacological actions suppress this immobility time.

Experiment result

Measurement of nerve cell protection effect

The cultured SH-SY5Y cells were divided into 96-well plates at a concentration of 2.5 × 10 ⁴ cells / well and cultured for 1 day. Then, 200 μM dexamethasone was added to induce cytotoxicity. (10 [mu] M, 50 [mu] M, 100 [mu] M) for 48 hours. Cell viability was confirmed by MTT method. That is, a solution (0.3 mg / mL final conc.) Of pMTT (3- [4,5-dimethyl-thiazol-2-yl] -2,5- diphenyltetrazolium bromide, Sigma Co., USA) After incubation for 4 hours, the culture medium was removed and 150 μL of DMSO was added to each well. The MTT formazan was dissolved for 5 minutes. After 5 minutes, the absorbance was measured with an ELISA reader (450 nm, 25 ° C.) Percentage of survival rate.

FIG. 1 is a graph showing the neuroprotective effect of Sansa sp. Extract. In the case of dexamethasone-added cell line (DEX), the survival rate was about 68% according to cytotoxicity. Survival rate, and the antidepressants imipramine and imipramine. Cell survival rate was significantly higher than that of the cell line treated with John's wort.

Float time and swimming time measurement

To measure the forced swimming behavior test (FST), a transparent beaker with a height of 300 cm or more was filled with 15 cm of water (temperature 2325 ° C) and the mice were swallowed for 6 minutes. Recorded for 6 minutes in a digital video file and analyzed for immobility and swimming time using a behavior analysis program (SMART v3.0, Panlab SL, Barcelona, Spain). Each result was analyzed for immobility time and swimming time while the mouse was not moving on the water surface for 4 minutes except for the first 2 minutes.

FIG. 2A and FIG. 2B are graphs showing the results of a forced swimming experiment after injecting the extract of Sansa sp. Extract into a mouse. As shown in the figure, the group administered orally with Sansha sp. Extract was a group The immobility time was significantly decreased and the swimming time was increased. In other words, it can be seen that the extract of Sansa sp.

  While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (4)

Crataegus The present invention relates to a pharmaceutical composition for the treatment or prevention of depression.
The pharmaceutical composition according to claim 1, wherein the horseshoe extract is extracted with water, a C ₁ to C ₄ lower alcohol, or a mixed solvent thereof.
Crataegus pinnatifida ) < / RTI > extract.
The food composition according to claim 3, wherein the composition is a powder, a granule, a tablet, a capsule or a beverage.

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