KR102167989B1 - Composition for preventing, ameliorating or treating infertility caused by implantation failure comprising ginseng extract as effective component - Google Patents

Abstract

The present invention relates to a composition for the prevention, improvement or treatment of infertility due to implantation disorders containing ginseng extract as an active ingredient, and in detail, the ginseng extract of the present invention is non-cytotoxic, and endometrial cells and trophoblast cell lines It has the effect of not only enhancing liver adhesion, increasing the expression of angiogenesis factors, but also significantly increasing the number of implanted embryos in implantation inhibition models, so that drugs for the prevention or treatment of infertility due to implantation disorders, or infertility due to implantation disorders It can be usefully used in health functional foods for the prevention or improvement of

Description

Composition for preventing, ameliorating or treating infertility caused by implantation failure comprising ginseng extract as effective component, containing ginseng extract as an effective component}

The present invention relates to a composition for preventing, improving or treating infertility due to implantation disorders containing ginseng extract as an active ingredient.

Infertility is defined as the case where a couple who did not use contraception do not reach pregnancy within one year even in a normal marital relationship. The cause of such infertility is infertility if there is an abnormality at any one stage of the entire process, from the development of male or female reproductive cells (sperm or egg) to fertilization of sperm and egg, development of a fertilized embryo, and implantation of the embryo in the uterus. Male factor, decreased ovarian function, ovulation disorder, fallopian tube injury, ligation (knot), or peritubular adhesion, cervix or immunological factor, uterine factor, immunological factor, severe systemic disease, unknown cause There are various causes such as.

Implantation is a phenomenon in early pregnancy in which the blastocyst, which is fertilized and developing into an embryo, attaches to the endometrium layer of the mother. Decidualization is induced in the endometrium layer of the mother, and is present at the edge of the blastocyst. As the differentiation of trophoblast cells rapidly progresses by cell fusion of cytotrophoblasts, they are differentiated into syncytiotrophoblasts, and stable implantation is achieved through interaction with endometrial cells. At this time, implanted syncytiotrophoblast cells undergo an invasive action known as a characteristic characteristic of trophoblast cells to the endometrial layer. Therefore, for the success of implantation, the first step for maintaining pregnancy, the invasion capacity of trophoblasts present in the blastocyst acts as an important factor. The infiltrated trophoblast cells infiltrate the maternal coronary artery existing at the base of the endometrium and change the vascular structure, thereby forming the vascular structure necessary for the development of the embryo, playing an important role in the development of the placenta as well as in maintaining a normal pregnancy. do. Therefore, improper invasive ability of trophoblast cells deformedly affects implantation disorder in the early pregnancy and development of blood vessels in the placenta, leading to placental insufficiency and vascular endothelial cell dysfunction, thus directly or indirectly, pregnancy maintenance, fetal growth disorders and placenta. It can also lead to obstetric diseases caused by the inappropriate development of

On the other hand, as a technology related to infertility, a method for manufacturing an herbal medicine composition for treating female infertility containing antlers is disclosed in Korean Patent No. 1600751, and the use of ovulation impregnated chamber extract having an implantation promoting effect and its use is disclosed in Korean Patent No. 1802675. It is disclosed, and Korean Patent Publication No. 2016-0118481 discloses a composition for improving ovarian aging containing a ginseng fruit extract, but preventing, improving or improving infertility due to implantation disorders containing the ginseng extract of the present invention as an active ingredient No therapeutic composition has been disclosed.

The present invention is derived from the above requirements, the present invention provides a composition for the prevention, improvement or treatment of infertility due to implantation disorders containing ginseng extract as an active ingredient, and ginseng extract as an active ingredient of the present invention By confirming that there is no cytotoxicity, it enhances the adhesion between endometrial cells and trophoblast cell lines, increases the expression of angiogenesis factors, and has the effect of significantly increasing the number of implanted embryos in the implantation inhibition model. The invention was completed.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating infertility due to implantation disorders containing ginseng extract as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving infertility due to implantation disorders containing ginseng extract as an active ingredient.

In addition, the present invention provides a method for promoting implantation in a uterus of a fertilized egg comprising administering a ginseng extract to mammals other than humans.

The present invention relates to a composition for the prevention, improvement or treatment of infertility due to implantation disorders containing ginseng extract as an active ingredient, and in detail, the ginseng extract of the present invention is non-cytotoxic, and endometrial cells and trophoblast cell lines It has the effect of increasing the liver adhesion, increasing the expression of angiogenic factors, and significantly increasing the number of implanted embryos in the implantation inhibition model.

1 is a result of confirming the cell viability of the ginseng extract of the present invention on ovarian cancer cells (TOV-21).
2 is a result of confirming the adhesion between cells of ginseng extract.
Figure 3 is a result of confirming the expression level of the neovascular factor PDGF according to the treatment of ginseng extract.
Figure 4 shows the increase in the number of implanted embryos after oral administration of ginseng extract in the implantation inhibiting animal model. ### indicates that the number of implanted embryos in the implantation suppression group (RU486) was statistically significantly reduced compared to the control group, p<0.001, and *** is ginseng of the present invention compared to the implantation suppression group (RU486) The number of implanted embryos in the extract-administered group (RU486 + ginseng) was statistically significantly increased, and p<0.001.

The present invention relates to a pharmaceutical composition for preventing or treating infertility due to implantation disorders containing ginseng extract as an active ingredient.

The ginseng extract may be prepared by a method comprising the following steps, but is not limited thereto:

(1) extracting ginseng by adding an extraction solvent;

(2) filtering the extract of step (1); And

(3) preparing an extract by concentrating the filtered extract in step (2) under reduced pressure and drying it.

In the step (1), the extraction solvent is preferably selected from water, a lower alcohol of C ₁ to C ₄ or a mixture thereof, and more preferably water (distilled water), but is not limited thereto.

In the above manufacturing method, all conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction, can be used as the extraction method. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried ginseng, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 110 ℃, more preferably 100 ± 2 ℃, but is not limited thereto. In addition, the extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but is not limited thereto. In the above method, the vacuum concentration in step (3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

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The pharmaceutical composition of the present invention may further include a carrier, excipient, or diluent in addition to the ginseng extract, and the pharmaceutical composition of the present invention may be administered orally or parenterally. , It is preferable to select intravenous, intramuscular, subcutaneous, intrauterine dura mater or cerebrovascular injection.

The pharmaceutical composition of the present invention may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose, or lactose ( lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, twin tween 61, cacao butter, laurin paper, glycero gelatin, and the like may be used.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, the'pharmaceutically effective amount' means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, and drug activity of the patient. , Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

The dosage of the composition of the present invention can be used in various ranges according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of disease.

In addition, the present invention relates to a health functional food composition for preventing or improving infertility due to implantation disorders containing ginseng extract as an active ingredient.

The health functional food composition is preferably prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrup, and beverages, but is not limited thereto, and may be prepared by adding it as an ingredient of food, and conventional It can be suitably manufactured according to the phosphorus method. Examples of foods to which the ginseng extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, It may be in any one form selected from tea, drink, alcoholic beverage, and vitamin complex, and includes all health functional foods in the usual sense.

The health functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective properties. It may contain colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, it may contain pulp for the manufacture of natural fruit juices and vegetable beverages. These components may be used independently or in combination. The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used.

In addition, the present invention relates to a method for enhancing intrauterine implantation of a fertilized egg comprising administering a ginseng extract to mammals other than humans. The administration target may be a mammal such as a human, preferably a mammal other than a human, but is not limited thereto.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for describing the present invention in more detail, and it is apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

Example 1. Preparation of ginseng extract

Distilled water of 10 times the weight of ginseng (1,420 g) was added, followed by reflux extraction at 100±2° C. for 3 hours. Then, after the sieve filter, the cotton filter was performed. The filtered solution was lyophilized to powder (yield: 12.82%).

Example 2. Cell Survival rate Confirm

In order to confirm the cytotoxicity of the ginseng extract prepared in Example 1, the ginseng extract was treated by concentration in the ovarian cancer cell line (TOV-21 cell line), and cultured for 24 hours to check the cell viability.

As a result, as disclosed in FIG. 1, the cytotoxicity of the ginseng extract was hardly observed up to a concentration of 500 µg/ml, and it was decided to perform cell-to-cell adhesion at a concentration of 50 µg/ml where no cytotoxicity was observed. .

Example 3. Endometrial cell line and Trophoblast Cell line in vitro Binding ability Confirm

In order to confirm the cell-to-cell adhesion efficacy of the human-derived trophoblast cell line (JAr cell line) and the endometrial cell line (Ishikawa cell line) in an in vitro model, Ishikawa cells per well were dispensed into a 6-well plate at 1.5×10 ⁶ cells/well. After about 24 hours, the ginseng extract was treated for 48 hours. After that, the fluorescently stained JAr cells were incubated with Ishikawa cells for 30 minutes, washed, and the number of JAr cells adhered to the Ishikawa cells was measured.

As a result, after ginseng extract treatment, the adhesion between cells was superior to that of Control without ginseng extract treatment (Fig. 2).

Example 4. Effect of expression of angiogenesis-related factors

After treatment and cultivation of 50㎍/㎖ of ginseng extract in TOV-21 cells, the effect of the ginseng extract on the expression of platelet-derived growth factor (PDGF), an angiogenesis-related factor, was investigated in real time PCR. ) Method.

As a result, as disclosed in FIG. 3, it was confirmed that the expression level of the neovascular factor, PDGF, was increased.

Example 5. Confirmation of efficacy in implantation inhibition model

C57BL/6N 6-week-old female mice were induced to RU486 and established as an implantation suppression model, and then the efficacy of the ginseng extract of the present invention was confirmed.

The treatment concentration was converted to an adult (60kg) to determine the daily intake, and was administered orally for 18 days. Ginseng extract was administered, PMSG (5 IU) was administered, and 48 hours later, hCG (5 IU) was administered. The uterus was excised 4 days after the male and female conjugation to confirm pregnancy.

-Adult (60kg): 8g×2=16g (daily intake)

-16g/60kg×yield of ginseng extract

-Ginseng (12.82% yield)

-Daily oral dose of mouse (20g): 0.684mg/100µl in physiological saline

As a result, as disclosed in FIG. 4, when the ginseng extract was treated, the number of implanted embryos was statistically significantly increased.

Claims (7)

A pharmaceutical composition for preventing or treating infertility due to implantation disorders containing ginseng water extract as an active ingredient. delete delete The pharmaceutical composition for preventing or treating infertility due to implantation disorder according to claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredient. Health functional food composition for preventing or improving infertility due to implantation disorders containing ginseng water extract as an active ingredient. The health functional food for preventing or improving infertility due to implantation disorders according to claim 5, wherein the composition is prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, and beverage. Composition. Intrauterine implantation of a fertilized egg comprising the step of administering ginseng water extract to mammals other than humans.

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