KR101742968B1

KR101742968B1 - Composition containing alpha-cyperone for improving pregnancy

Info

Publication number: KR101742968B1
Application number: KR1020160012883A
Authority: KR
Inventors: 하기태; 최희정
Original assignee: 부산대학교 산학협력단
Priority date: 2016-02-02
Filing date: 2016-02-02
Publication date: 2017-06-15

Abstract

The present invention provides a composition for promoting pregnancy or an agent for the prevention or treatment of gynecological infertility comprising alpha-cipherone as an active ingredient. The alpha-cyperone of the present invention has an excellent effect of increasing the expression of LIF associated with the implantation in the endometrial cells and increasing the binding force between the endometrial cells and the fetal-derived trophoblast to effectively induce the implantation of the embryo, It can be useful in fields related to pharmaceuticals and functional foods that can prevent and treat gynecological infertility including implantation infertility.

Description

A composition for promoting pregnancy comprising alpha-cyperone as an active ingredient {Composition containing alpha-cyperone for improving pregnancy}

The present invention provides a composition for promoting pregnancy or an agent for the prevention or treatment of gynecological infertility comprising alpha-cipherone as an active ingredient.

In recent years, in conjunction with an aging society, infertility is increasing as maternal age increases. In addition, due to environmental pollution caused by industrialization and the advancement of women into society, the stress of women is increasing, and the pregnancy rate is significantly lowered. Common causes of female infertility include ovulation disorders, transfer of fertilized eggs, and implantation disorders. Infertility due to obstructive ovarian failure and transfer of embryos is largely solved through in vitro fertilization (IVF), but sterilization due to implantation disorder has yet to be clarified. Normal pregnancy progresses in the order of fertilization, implantation and fetal development. Quality of fertilized eggs and receptivity of endometrium are very important factors in embryo implantation. The major factors controlling the endometrium's water solubility include growth factors such as cytokine such as leukemia inhibitory factor (LIF), insulin-like growth factor-2 and interleukin-11 (IL-11) is important (Hum Rep Update, 12 (6): 731-746). Among them, the expression of LIF, an interleukin-6 (IL-6) family cytokine, is known to be a key factor controlling the implantation process. In particular, in mice deficient in LIF, there are reports of defects in the adhesion process between the embryo and the endometrium and conception and pregnancy (Nature 359 (6390): 76-79; Endocrinology 150 6): 2915-2923). In addition, it has been reported that inhibition of LIF function by using an antagonist for LIF does not result in implantation (PNAS 104 (49): 19357-19362). LIF is known to be the most important factor in the implantation process have.

Alpha-cyperone alleviates pulmonary cell damage caused by Staphylococcus aureus (J Microbiol Biotechnol. 22 (8): 1170-6), algae colicosis in chicken alveolar cells (Vet Immunol Immunopathol. 159 (1-2): 50-7) have been reported. It has also been reported that the alpha-cipherone is effective for anti-inflammatory activity in macrophages (J Ethnopharmacol. 147 (1): 208-14). However, no previous studies have shown that alpha-cyperone is effective in preventing or treating infertility and in pregnancy.

Therefore, the present inventors have found that alpha-cipheron increases the expression of LIF, a key cytokine related to the implantation in endometrial cells, and induces endometrial cells and fetal origins The present inventors have confirmed that the present invention is effective in promoting pregnancy and infertility caused by oocyte non-implantation by increasing the binding of trophoblast cells.

It is an object of the present invention to provide a composition for promoting pregnancy comprising alpha-cyperone as an active ingredient.

It is also an object of the present invention to provide a composition for the prevention or treatment of gynecological infertility comprising alpha-cyperone as an active ingredient.

In order to solve the above problems, the present invention provides a pharmaceutical composition for promoting pregnancy comprising alpha-cyperone as an active ingredient.

The present invention also provides a food composition for promoting pregnancy comprising alpha-cipherone as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating gynecological infertility comprising alpha-cyperone as an active ingredient.

The present invention also provides a food composition for preventing or ameliorating gynecological infertility comprising alpha-cyperone as an active ingredient.

The alpha-ciperone of the present invention has an excellent effect of increasing the expression of LIF associated with the implantation in the endometrial cells, increasing the binding force between the endometrial cells and the fetal-derived veterinary membrane cells and effectively inducing the implantation of the embryos. Therefore, the alpha-cyperone according to the present invention can be usefully used in fields related to medicines and functional foods for prevention and treatment of gynecological infertility including ovarian non-implantation infertility.

FIG. 1 is a graph showing the results of comparing the expression ratios of LIF mRNA of a single compound of alpha-cyperone and herbal suppositories.
Figure 2 shows the formula of alpha-cyperone.
FIG. 3 shows the results of confirming cytotoxicity according to alpha-cyperone treatment concentration. FIG.
FIG. 4 is a graph showing the results of RT-PCR for the increase in the expression of LIF mRNA according to the alpha-cyperone treatment concentration.
FIG. 5 is a graph showing the result of Western blot analysis of the increase in the expression of LIF protein according to the treatment concentration of alpha-ciperone.
Fig. 6 is a graph showing the result (A) of fluorescence microscopy and the result (B) of counting the number of adherent cells in Ishikawa cells treated with alpha-cyperone and a control group without treatment.
Fig. 7 is a graph showing the results of counting the number of adhesion of sperm cells to Ishikawa cells according to the treatment concentration of alpha-cyperone and herbal extract. Fig.

The present invention provides a composition for promoting pregnancy comprising alpha-cyperone as an active ingredient.

In the present invention, the "alpha-cyperone" is a component for which excellent antioxidative and anti-inflammatory effects have been reported. C ₁₅ H ₂₂ O ₂ , and has a molecular weight of about 234, which is shown in FIG. 2 of the present invention.

The alpha-cipheron increases the expression of leukemia inhibitory factor (LIF), but is not limited thereto.

The alpha-cepheone can be treated at a concentration of 1 to 90 [mu] M, but preferably at a concentration of 30 to 60 [mu] M. When alpha-cyperone is within the above-mentioned range, it is more preferable because it has no cytotoxicity.

In the present invention, "pregnancy" is a process in which the fertilized egg is implanted in the inner wall of the uterus, is supplied with nutrition from the mother, and develops into the fetus. The normal pregnancy process proceeds in the order of fertilization, implantation and fetal development.

The term "fertilization" means that an ovum ovulated in a female ovary meets with spermatozoa at the upper part of the oviduct to form an embryo. The ovum has the ability to stay alive for 1-2 days after ovulation and the sperm to remain alive for 2-3 days in the uterus, and the sperm survive for one week after being ejaculated. Sperm are spermatozoa that reach the ovary close to the ovary 2 ~ 3 hours after the ejaculation. In general, the spermatozoon first reaches the upper part of the oviduct, and when ovum is ovulated from the ovary, only one of the spermatozoa .

The "implantation" refers to a phenomenon in which an embryo in which a sperm and an egg are brought together at the upper part of the oviduct is moved to the uterus repeatedly and buried in the uterine wall in a bladder state. After the embryo is conceived, it is called pregnancy. Embryos implanted into the uterus are born after about nine months of age, while being fed from the uterine lining.

According to one embodiment of the present invention, alpha-cipherone increases the expression of leukemia inhibitory factor (LIF) in a cell than a single compound of the herbal supplement, and the binding force between endometrial cells and fetal-derived trophoblast cells Which is effective in inducing fertilization of embryos effectively.

The composition comprises a pharmaceutical composition or a food composition.

The composition may contain alpha-cyperone alone or a substance effective for promoting pregnancy as an active ingredient. In addition to the above-mentioned active ingredients, the composition may further contain additional components such as pharmaceuticals Or nutritional acceptable carriers, excipients, diluents or subcomponents.

More specifically, the pharmaceutical composition for accelerating pregnancy comprising the alpha-ciperol may further contain, in addition to the active ingredient, a nutritional supplement, a vitamin, an electrolyte, a flavoring agent, a colorant, a heavy stabilizer, a pectic acid and a salt thereof, Organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, and carbonating agents used in carbonated beverages.

Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium stearate, mineral oil, calcium carbonate, dextrin, propyleneglycol, liquid paraffin, sodium carboxymethylcellulose, sodium carboxymethylcellulose, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, And physiological saline, but is not limited thereto. The components can be added to the active ingredient, i.e. alpha-cepheone, either independently or in combination.

The composition according to the present invention can be used alone for promoting pregnancy, or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

The composition according to the present invention may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically) according to a desired method, and the dosage may be appropriately selected according to the weight, age, sex, The range varies depending on the condition, diet, time of administration, method of administration, excretion rate and severity of the disease. The daily dose of alpha-ciperol of the present invention is 0.01 to 10,000 mg / kg, preferably 1 to 20 mg / kg, and is preferably administered once to three times a day.

The composition according to the present invention may be included in a health food for the purpose of promoting pregnancy. When the active ingredient of the present invention is used as a food additive, the composition isolated from the synthesis or the extract may be added as it is, And can be suitably used according to a conventional method. The amount of the active ingredient to be mixed may be appropriately adjusted depending on the intended use (prevention, health or therapeutic treatment).

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, , Alcoholic beverages and vitamin complexes, and includes all health foods in a conventional sense.

Various additives such as flavors or natural carbohydrates can be used as the food-aid additive of the present invention. The above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

In addition to the above, the composition according to the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid concentrating agents, pH adjusting agents, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the composition according to the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination.

The present invention also provides a composition for the prevention or treatment of gynecological infertility comprising alpha-cyperone as an active ingredient.

The composition comprises a pharmaceutical composition or a food composition.

The female infertility is related to nonimplantation of ovum, female infertility associated with anovulation associated with anovulation, female infertility of tubal origin, Tubal occlusion, Tubal stenosis, Female infertility of the uterine origin, Infertility of the uterus, Infertility of the uterus, Infertility associated with male factors, female infertility associated with cervical origins, female infertility associated with male factors, and other factors related to congenital anomaly of uterus. At least one selected from the group consisting of female infertility of other origin and unspecified female infertility (unspecified) But is not limited thereto.

In the present invention, the above-mentioned "Nonimplantation of ovum" is a representative cause of female infertility. During fertilization, the embryo moves while dividing cells into the uterus, and when it reaches the blastocyst stage, it is buried in the endometrium and seated. However, when embryos are not thick enough to be implanted or stiff due to damage, it is difficult to obtain implants, and abortion is often caused by lack of space due to adhesion of the uterine wall. Endometrial hyperplasia due to intrauterine inflammation caused by spontaneous abortion, mastectomy, abortion, endometritis, pelvic tuberculosis and intrauterine device, or uterine myopathy, Or if the uterine endometrium is incomplete due to congenital abnormal uterine hormone secretion. The oocyte implantation has been identified as the major cause of artificial insemination and failure of in vitro procedures.

Since the pharmaceutical composition or the food composition includes a pharmaceutical preparation or a food preparation containing the above-mentioned alpha-cipheron as an active ingredient, the contents overlapping with the composition of the present invention described above can be seen by the overlapping description The description thereof is omitted in order to avoid excessive complexity of the specification.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be obvious to you.

Example One. Herbalist Cyperus rotundus L.) of For a single compound LIF mRNA Increased expression and selection of active ingredients

The increase of mRNA expression of leukemia inhibitory factor (LIF), which is known to be important factor in the process of implantation, was confirmed for a single compound of herbal supplement and an effective ingredient related to the implantation was selected from the single compound.

Specifically, a single compound of the perfume, ferulic acid, humulene, luteolin, protocatechuic acid, vanillic acid, p-cymene, caffeine, Caffeic acid, coumaric acid, alpha-cyperone and nootkatone were each purchased from Sigma. Ishikawa cells were treated with the single compound at a concentration of 50 μM and cultured for 24 hours. Then, RNA was extracted and the amount of mRNA expression was confirmed by RT-PCR using LIF-specific primers. The primer sequence used for the amplification of LIF used for PCR amplification was 5'-GGCCCGGACACCCATAGACG-3 '(SEQ ID NO: 1) in the forward direction and 5'-CCACGCGCCATCCAGGTAAA-3' (SEQ ID NO: 2) The primer sequence used for amplification of actin is 5'-CAAGAGATGGCCACGGCTGCT-3 '(SEQ ID NO: 3) in the forward direction and 5'-TCCTTCTGCATCCTGTCGGCA-3' (SEQ ID NO: 4) in the reverse direction. Each amplified DNA was electrophoresed and photographed under ultraviolet light (UV). The intensity of the band was measured using a densitometer, and the ratio of the expression level of LIF and the expression level of? -Actin, which is a control standard, Respectively.

As shown in FIG. 1, it was confirmed that the expression ratio of LIF, which is known to be an important factor in the implantation process, is higher than that of other herbal extracts, and it is selected as an active ingredient and a series of experiments .

Example 2. Alpha- Cyperon Cytotoxicity Assessment

The toxicity of alpha-cyperone to cells was confirmed by MTT (3- (4,5-dimethylthiazole-2-yl) -2,5-diphenyltetrazoli-umbromide) assay.

Specifically, the endometrial cell line, Ishikawa cell, was treated with alpha-cipheron having the formula of Fig. 2 at concentrations of 0, 5, 10, 30, 50 and 100 μM for 24 hours, respectively. The MTT solution (2 mg / ml) was reacted at 37 ° C for 4 hours in a CO ₂ incubator and the supernatant was removed. Live cells were reduced by intracellular redox enzyme, and purple dyes (Formazan) were dissolved in DMSO and measured at 540 nm wavelength with a microplate reader. Live cells were expressed as percentage compared to the control group. The results are shown in Fig.

As shown in FIG. 3, it was confirmed that no significant cytotoxicity was confirmed even when alpha-cyperone was treated to a concentration of 50 μM. Therefore, the concentrations below 50 μM without toxicity were used in the experiments below.

Example 3. Alpha- Cyperon Leukemia inhibitory factor (leukemia inhibitory factor; LIF ) mRNA Confirmation of expression of

The expression level of LIF mRNA according to the treatment concentration of alpha-cyperone was confirmed by RT-PCR.

Specifically, Ishikawa cells were treated with alpha-cipheron at concentrations of 0, 10, 30 and 50 μM and cultured for 24 hours. Then, the RNAs of the cells treated at the respective concentrations were extracted and primers specific for LIF were used. The sequence of the primer was 5'-GGCCCGGACACCCATAGACG-3 '(SEQ ID NO: 1) in the forward direction and 5'-CCACGCGCCATCCAGGTAAA-3' (SEQ ID NO: 2) in the reverse direction. The primer sequence used for amplification of- 3 '(SEQ ID NO: 3) and 5'-TCCTTCTGCATCCTGTCGGCA-3' (SEQ ID NO: 4) in the reverse direction. CDNA was synthesized by total RNA, oligo-dT primer and M-MLV reverse transcriptase at 42 ° C for 1 hour. The cDNA was amplified by 30 cycles of denaturation at 95 ° C. for 30 seconds, annealing at 60 ° C. for 30 seconds, and extension at 72 ° C. for 30 seconds using the primers described above. And the amount of mRNA expression was confirmed by polymerase chain reaction. The amplified DNA was electrophoretically photographed under ultraviolet light (UV) and the intensity of the band was measured using a densitometry. In addition, the expression level of LIF mRNA and the ratio of expression amount of? -Actin mRNA as a control standard were confirmed. The results are shown in Fig.

As shown in FIG. 4, when the endometrial cells were treated with alpha-cipheron at 50 μM, the expression of LIF mRNA was 1.82 times that of the control.

Therefore, it was confirmed that the expression of LIF mRNA was increased by treatment with alpha-cyperone .

Example 4. Alpha- Cyperon Depending on treatment concentration LIF Identification of Protein Expression

Expression of LIF protein was confirmed by western blotting according to the treatment concentration of alpha-cyperone.

More specifically, Ishikawa cells were treated with alpha-cyperone at a concentration of 0, 10, 30 and 50 μM, and cultured for 24 hours. Proteins were extracted from the cells and electrophoresed by SDS-PAGE. Subsequently, the separated proteins were transferred to a nitrocellulose membrane by electrophoresis and then sequentially bound to LIF-specific antibodies (SantaCruz) and secondary antibodies to which HRP (Horseradish peroxidase) had been added ECL (enhanced chemiluminescence) solution. The intensity of the band was measured using a densitometer and the ratio of the expression amount of LIF protein and the expression amount of GAPDH protein as a representative standard was confirmed. The results are shown in Fig.

As shown in FIG. 5, when the endometrial cells were treated with 10, 30 and 50 μM of alpha-cyperone, LIF protein expression was increased to about 1.18, 2.52 and 2.70 times as compared with the control group.

Therefore, it was confirmed that the expression of LIF protein is increased depending on the concentration of alpha-cyperone.

Example 5. Alpha- Cyperon Of the JAR cells to the treated cells Attachment Confirm

Adhesion of embryonic stem cells, a fetal-derived trophoblast cell line, to Ishikawa cell line derived from endometrium treated with alpha-cyperone was confirmed.

Specifically, human endometrial cell line Ishikawa cells were cultured in a single layer so as to have an 80% confluence, treated with alpha-cipheron to a concentration of 50 μM, and cultured for 48 hours to obtain 100% To become a cluster. Subsequently, the pre-cultured fetal-derived cell line was removed from the culture dish, and the cell was stained well to be a single cell and stained with CellTracker ^™ Green CMFDA (5-chloromethylfluorescein diacetate), a cell permeable green fluorescent substance . Thereafter, the cells were sprinkled on a single layer of Ishikawa cells and cultured in an incubator at 37 캜 for 30 minutes while stirring at a speed of about 60 rpm. The cultured cells were washed twice with phosphate buffered saline (PBS) to remove unattached cells, and the adhered cells were fixed with 3.7% formalin solution. When confirmed by fluorescence microscopy, the number of cells that fluoresce by CMFDA was the number of spermatocytes attached to the Ishikawa cells at the bottom, and was photographed (Fig. 6 (A)). Thereafter, the total number of cells counted in photographs of five or more different regions was compared with a control group not treated with alpha-cyperone, and then the mean and standard deviation of the values repeated three times were determined (FIG. 6 B)). The results are shown in Fig.

As shown in Figs. 6 (A) and 6 (B), the adherence of autologous cells to Ishikawa cells treated with alpha-cyperone was significantly more than twice as high as that of the control without alpha-cyperone Respectively.

Example 6. Alpha- Ciperon And Herbalist extract( Cyperus rotundus ) Of cells treated with JAR cells Attachment Confirm

In Example 5, it was found that alpha-cyperone increased the adherence of the embryo-derived trophoblast cell line to the endometrial Ishikawa cell line, and the adherence of the alpha-cyperone and herbal extracts of the present invention Respectively.

Specifically, Ishikawa cells, a human endometrium-derived cell line, were cultured in a single layer so as to have an 80% confluence. Then, the cells were treated with 10 μg of alpha-cyperone or herbal extract and cultured for 48 hours to obtain 100% To become a cluster. Subsequently, the pre-cultured fetal-derived cell line was removed from the culture dish, and the cell was stained well to be a single cell and stained with CellTracker ^™ Green CMFDA (5-chloromethylfluorescein diacetate), a cell permeable green fluorescent substance . Thereafter, the cells were sprinkled on a single layer of Ishikawa cells and cultured in an incubator at 37 캜 for 30 minutes while stirring at a speed of about 60 rpm. The cultured cells were washed twice with phosphate buffered saline (PBS) to remove unattached cells, and the adhered cells were fixed with 3.7% formalin solution. Subsequently, the number of sperm cells attached to the Ishikawa cells on the bottom were measured immediately after the fluorescence by CMFDA. The results are shown in Fig.

As shown in Fig. 7, it was confirmed that the adherence ability of spermatocytes to Ishikawa cells treated with alpha-cyperone was about 2 times higher than that of the control group.

Therefore, the alpha-ciperone of the present invention was found to have a higher rate of increase in expression of LIF mRNA, which is an important factor in the process of implantation, than that of the single conjugate compound, and that the adherence of sister cells to Ishikawa cells is superior to that of the herbal extract- As an effective active ingredient, not only promoting pregnancy but also effects on gynecological infertility can be expected.

Hereinafter, the pharmaceutical composition of the present invention and the preparation example of the food composition will be described, but the present invention is not intended to be limited but is specifically described.

Formulation example 1. Preparation of pharmaceutical compositions

1-1. Sanje Produce

Alpha-cyperone 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

1-2. Manufacture of tablets

Alpha-cyperone 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

1-3. Preparation of capsules

Alpha-cyperone 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

1-4. Injection preparation

Alpha-cyperone 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na2HPO42H2O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

1-5. Liquid Produce

Alpha-cyperone 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example 2. Preparation of food composition

2-1. Health food manufacturing

100 mg of alpha-cyperone, a proper amount of vitamin mixture, 70 g of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.2 mg of vitamin B6, 0.2 g of vitamin B12, 10 mg of vitamin C, Amide 1.7 mg, folic acid 50 g, calcium pantothenate 0.5 mg, inorganic mixture suitable amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium phosphate monohydrate 15 mg, dicalcium phosphate 55 mg, potassium citrate 90 Mg of calcium carbonate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride were mixed and granules were prepared and a health food was prepared according to a conventional method. At this time, although the composition ratio of the vitamin and mineral mixture is relatively mixed with the ingredient suitable for health food, it may be arbitrarily modified.

2-2. Health drink manufacturing

100 g of vitamin C, 15 g of vitamin C, 100 g of vitamin E (powder), 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 0.2 g of vitamin A, 0.25 g of vitamin B1 , 0.3 g of vitamin B2 and a predetermined amount of water were mixed and heated at 85 DEG C for about 1 hour with stirring. The resulting solution was filtered to obtain a sterilized 2 L container, sealed sterilized and refrigerated to prepare a health drink. At this time, although the composition ratio of the ingredients suitable for the beverage is comparatively mixed, the mixture ratio may be arbitrarily varied according to the demand, the demanded country, the intended use, and the regional or national preference.

<110> Pusan National University Industry-University Cooperation Foundation <120> Composition containing alpha-cyperone for improving pregnancy <130> 1-262 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> forward primer for LIF <400> 1 ggcccggaca cccatagacg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for LIF <400> 2 ccacgcgcca tccaggtaaa 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for alpha-actin <400> 3 caagagatgg ccacggctgc t 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for alpha-actin <400> 4 tccttctgca tcctgtcggc a 21

Claims

A pharmaceutical composition for promoting pregnancy comprising alpha-cyperone as an active ingredient.

The method according to claim 1,
The pharmaceutical composition for promoting pregnancy is characterized in that the alpha-ciperone increases the expression of a leukemia inhibitory factor (LIF).

The method according to claim 1,
Wherein the alpha-cepheone is treated at a concentration of 1 to 90 [mu] M.

A food composition for promoting pregnancy comprising alpha-cyperone as an active ingredient.

A pharmaceutical composition for the prevention or treatment of gynecological infertility comprising alpha-cyperone as an active ingredient.

6. The method of claim 5,
The gonadal infertility can be classified into three categories: nonimplantation of ovum, female infertility associated with anovulation, female infertility of tubal origin originating from the uterine tube, female infertility originating from the uterus wherein the female infertility is associated with at least one of at least one selected from the group consisting of infertility of uterine origin, female infertility of cervical origin originating from the cervix and female infertility associated with male factors. A pharmaceutical composition for preventing or treating.

A food composition for prevention or improvement of gynecological infertility comprising alpha-cyperone as an active ingredient.