KR101582734B1 - Composition for preventing or treating uterine leiomyoma comprising flavopiridol - Google Patents

Abstract

The present invention relates to a composition for preventing or treating uterine leiomyoma containing flavopiridol as an active ingredient, and more particularly, to a composition for preventing or treating fibroids in uterine leiomyoma cells, The present invention provides a pharmaceutical composition or a health food composition capable of preventing or treating uterine leiomyoma through inhibition of cell cycle progression and induction of apoptosis through reduction of proliferation and cell viability of uterine leiomyoma cells.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating uterine leiomyoma comprising flavopiridol as an active ingredient,

The present invention relates to a composition for preventing or treating uterine leiomyoma by inhibiting cell cycle progression of uterine leiomyoma cells by containing flavopiridol as an active ingredient and inducing cell death.

Flavopyridone is the first flavonoid derivative isolated from the stem skin of Dysoxylum binectariferum, the first clinically developed substance to be a pan inhibitor of cyclin dependent kinase (CDK). Flavopyridone competes directly with APT and can inhibit cell cycle progression and induce apoptosis by inducing G1 blockade by inhibiting CDK2, CDK4, and CDK6 and negative PI3K / AKT signaling pathways.

Flavopyridone with these characteristics has been studied as a therapeutic agent for various diseases, but the effect on the prevention or treatment of fibroids using Flavopyridone has not been reported.

The uterine myoma is commonly referred to as myoma, fibroma, or fibroid, and is a smooth muscle tumor originating from the myometrium. It occurs in more than 40% of women of reproductive age, and the incidence rate of women after their 50s is 70-80% It is a rapidly growing disease and is currently being mentioned as one of the major diseases of interest. Patients with uterine leiomyomas present with medically symptomatic symptoms such as anemia, menstrual hyperpigmentation, pelvic pain, infertility, spontaneous abortion, abnormal uterine bleeding, and addictive abortion. Some medical procedures may be applicable to women who wish to preserve the uterus, The main treatment method of myoma is surgical operation.

Recently, gonadotropin-releasing hormone (GnRH) antagonists have been used to reduce the degree of ovarian hormone circulation in the management of uterine myoma, but GnRH antagonists, such as the feeling of skin feel the menopausal women Is a major cause of side effects, and long-term use of these drugs can cause osteoporosis and cardiovascular disease.

Therefore, it is necessary to study safe and effective treatment methods without surgery for uterine leiomyoma patients.

Korean Patent Laid-Open No. 10-2007-7009411

In order to solve the above problems, the present invention provides a composition containing flavopiridol as an active ingredient, thereby solving the problem of side effects of drugs used in conventional uterine myoma treatment, And to provide an effective treatment method capable of preserving the uterus through treatment using the same.

The present invention provides a composition for preventing or treating uterine myoma that contains flavopiridol as an active ingredient.

The Flavopyridone selectively inhibits cell cycle progression of uterine leiomyoma cells, thereby inhibiting cell proliferation and inducing cell death.

The present invention also provides a pharmaceutical composition for preventing or treating uterine leiomyoma containing flavopiridol as an active ingredient.

The pharmaceutical composition may contain 0.01 to 10 parts by weight of flavopyridone, based on 100 parts by weight of the total amount.

In another embodiment, the present invention provides a health food composition for improving or preventing uterine myoma that contains flavopiridol as an active ingredient.

The composition containing the flavopiridol according to the present invention as an active ingredient selectively acts only on uterine myoma cells to inhibit the expression of proteins related to the cell cycle of uterine myoma cells, thereby inhibiting cell cycle progression and inhibiting cell proliferation By inducing cell death, uterine myoma can be prevented or treated without affecting normal myometrial cells.

Figure 1 shows the percentage of cells surviving after 24 hours of treatment with Flavopyridone as a result of MTT analysis showing the effect of Flavopyridol on uterine leiomyoma cell proliferation, * P <0.05.
FIG. 2 shows that Flavopyridone inhibited the DNA synthesis of fibroblastoma cells. As a result, Flavopyridone was treated with 50, 100, 150 and 200 nM for 24 hours and labeled with 10 μM BrDU for 24 hours. Expressed as a relative value to the BrDU percentage of control cells, * P < 0.05.
FIG. 3 shows the results of cell flow analysis after the treatment with Flavopyridone for 24 hours as a result of Flavopyridol-treated cell cycle. The results are as follows: the percentage of whole cells, .
FIG. 4 shows the effect of Flavopyridone on the expression of proteins related to the cell cycle of uterine leiomyoma cells. After the treatment with Flavopyridone for 24 hours, proteins were extracted from the cells and subjected to immunoblot analysis. As a control, β - actin was used.
FIG. 5 shows the results of histological evaluation through hematoxylin and eosin staining after human-derived uterine myoma xenotransplantation and the use of antibodies against α smooth muscle actin, desmin, estrogen receptor α (ER) and progesterone receptor (PR) The result of immunochemical analysis on the model and cell transplantation model, each photograph was × 200 times.
FIG. 6 shows the effect of Flavopyridone administration on xenografted tumors. As a result, A was administered to the xenografted animal model in an experimental group in which 5.0 or 7.5 mg / kg of Flavopyridol was administered for 3 weeks and a control group in which DMSO was administered , B is the photograph of the tumor formed in each experimental group and the control group, C is the weight of the tumor after stopping the drug treatment of each of the flavopiride and DMSO control groups , And * P &lt; 0.05.

The present invention provides a composition for preventing or treating uterine myoma that contains flavopiridol as an active ingredient.

The Flavopyridone selectively inhibits cell cycle progression of uterine leiomyoma cells, thereby inhibiting cell proliferation and inducing cell death.

More specifically, the results of MTT assay of the cell viability of the experimental group treated with Flavopiridol and the control group treated with DMSO showed that Flavopyridone The number of uterine myoma cells decreased in a dose dependent manner. Especially, the survival rate of uterine myoma cells was reduced by more than 50% in the experimental group treated with 200 nM Flavopyridone. On the other hand, when Flavopyridone was treated with normal uterine muscle cells , There was no significant decrease in normal myometrial cells.

As a result of the colorimetric analysis conducted to confirm the ability of Flavopyridone to proliferate myomaoma cells, it was confirmed that the DNA synthesis of fibroblast-treated uterine myoma cells was reduced as shown in FIG. 2, Similar to the results, it was confirmed that the proliferation of uterine leiomyoma cells was suppressed.

To investigate the effect of Flavopiridol on myoma cells, FACS and Western blot analysis were performed on 100 or 200 nM Flavopyridol-treated experimental group. Flow cytometric analysis As shown in Fig. 3, which is a result of FACS, it was confirmed that the period of uterine myoma cells in the experimental group treated with flavopiridol stopped at the G1 phase.

Also, as shown in Western blot in FIG. 4, expression of CDK inhibitors p21 ^{cip / wafl} and p27 ^kip1 was increased while expression of CDK2, CDK4 and cyclin A was decreased.

From the above results, it was confirmed that Flavopyridone selectively inhibited the expression of CDK gene in uterine leiomyoma cells, thereby inhibiting cell cycle progression and inducing cell death, thereby reducing proliferation and cell survival of uterine leiomyoma cells.

The present invention also provides a pharmaceutical composition for preventing or treating uterine leiomyoma containing flavopiridol as an active ingredient.

The pharmaceutical composition may contain 0.01 to 10 parts by weight of flavopyridone, based on 100 parts by weight of the total amount.

In one embodiment of the present invention, the pharmaceutical composition for preventing or treating uterine leiomyoma containing the above-mentioned flavopiridol as an active ingredient may be administered orally or parenterally in the form of injections, granules, powders, tablets, pills, capsules, suppositories, Any one of the formulations selected from the group consisting of suspensions, emulsions, drops, and liquid preparations can be used.

In another embodiment of the present invention, the pharmaceutical composition for preventing or treating uterine myoma of the present invention containing flavopiridol as an active ingredient may be formulated into an appropriate carrier, excipient, disintegrant, sweetener, , At least one additive selected from the group consisting of lubricants, lubricants, flavors, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, binders and lubricants.

Specific examples of carriers, excipients and diluents include 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. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

According to one embodiment of the present invention, the pharmaceutical composition may be administered orally, intraarterally, intraperitoneally, intramuscularly, intraarterally, intraperitoneally, intrasternally, transdermally, nasally, inhaled, topically, rectally, &Lt; / RTI &gt; can be administered to the subject in a conventional manner.

The desired dosage of the flavopiridol may vary depending on the condition and body weight of the subject, the kind and degree of the disease, the drug form, the administration route and the period, and may be appropriately selected by those skilled in the art. According to one embodiment of the present invention, the daily dose may be 0.01 to 200 mg / kg, specifically 0.1 to 200 mg / kg, more specifically 0.1 to 100 mg / kg, though it is not limited thereto. The administration may be performed once a day or divided into several times, and thus the scope of the present invention is not limited thereto.

In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.

In another embodiment, the present invention provides a health food composition for improving or preventing uterine myoma that contains flavopiridol as an active ingredient.

The health food may be provided in the form of powder, granules, tablets, capsules, syrups or beverages. The health food may be used in combination with other food or food additives in addition to the active ingredient flavopiridol according to the present invention, May be appropriately used depending on the method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The effective dose of the flavopiridol contained in the health food may be used in accordance with the effective dose of the pharmaceutical composition. However, in the case of long-term intake intended for health and hygiene purposes or health control purposes, And it is clear that the active ingredient can be used in an amount exceeding the above range since there is no problem in terms of safety.

There is no particular limitation on the type of the health food, and examples thereof include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, Drinks, alcoholic beverages and vitamin complexes.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Collecting uterine myoma

Hysterectomy and adjacent normal uterine myometrium were obtained by hysterectomy. Hysterectomy was performed at Keumyong University Dongsan Medical Center. The consent of the patient was obtained from the patient and the collection and processing of the sample was conducted according to the guidance of the ethics committee of the Dongsan Medical Center of Keimyung University.

The uterine leiomyoma was collected from the center to the periphery of the largest fibroma of the women who underwent hysterectomy for the leiomyoma selectively. The central part of the collected tissue was restricted to a volume of 5 mm and the periphery was seen from the capsule to the outer 5 mm. All myometrial leiomyomas were calcified, including hydromyxoid myomas.

Patients were women aged 40-48 years, with an average age of 45 years. Three samples of the menstrual cycle phase 3 and secretory phase of the menstrual cycle were collected from patients without hormone therapy for at least 6 months before the operation.

The fibrous tissue was collected aseptically, immediately immersed in phosphate-buffered saline (PBS) containing 100 units / mL of antibiotics, and transferred to the laboratory in the presence of ice.

< Example  2> Primary uterine leiomyoma cell culture

Tissues were washed twice with cold PBS and cut into 5 mm pieces in a sterile culture dish. The slices were incubated in Hanks' supplemented with 25 mM HEPES, 100 units / mL of antibiotic, 1.5 mg / mL collagenase (Sigma-Aldrich, USA) and 0.2 mg / mL DNase I (Roche Diagnostics, Mannheim, Germany) Were transferred to a 50 mL conical tube containing balanced salts (HBSS; Sigma-Aldrich, St Louis, MO, USA).

All tubes were kept in a constant temperature bath at 37 占 폚 and stirred for 3 hours.

Undigested tissue was filtered and centrifuged at 1000 rpm for 5 minutes. Cell pellets were washed with HBSS and dispersed in a medium consisting of 10% fetal bovine serum (FBS), DMEM / F12 and 100 units / mL of antibiotics.

In order to analyze the characteristics of the uterine leiomyoma cells and to confirm the accuracy of the cells after the cell separation process, non-mentin antibody, which is an intermediate light filament protein present in the fibroblasts, was used. Further, desmin staining revealed that the smooth muscle cells .

< Example  3> cells Survival  analysis

To analyze the cell viability, colorimetric 3- (4,5-dimethylthiazol-2-yl) -2,5-dimethylthiazol- -2,5-diphenyltetrazolium bromide (MTT) analysis.

First, normal myometrial cells and uterine leiomyoma cells were plated at 1 × 10 ⁵ cells / mL per well in a 24-well plate and cultured until each cell was attached to the plate. After cell attachment, Flavopiridol 50, 100, 150, and 200 nM were replaced with media containing DMSO as a control and cultured for 24 hours. The MTT reagent was then added and the optical density (OD) was measured at 495 nm by incubation at 37 ° C for an additional 3 hours.

The cell viability was normalized by dividing the OD value of the test well by the OD value of the control well after subtracting the background OD value of the wells containing the wells, and the significance and standard deviation were calculated by repeating the measurement three times.

As a result of analysis of the effect of Flavopyridone on normal myometrium and uterine leiomyoma cell growth by MTT analysis, the survival of cells was decreased dose-dependently in Flavopyridol-treated experimental group as shown in Fig. In particular, it was confirmed that cell survival was reduced by more than 50% in the experimental group treated with 200 nM flavopiridol.

In addition, when the normal uterine myocardial cells were treated with Flavopyridol, there was no decrease in cell growth. From the above results, it was confirmed that Flavopyridone has no effect on normal myometrial cells but has an effect of inhibiting the growth of uterine myoma cells.

< Example  4> Cell proliferation assay

The ability of Flavopyridone to inhibit DNA synthesis was assessed by evaluating the amount of 5-bromo-2'-deoxyuridine (BrDU) contained in the DNA through colorimetric immunoassay.

The colorimetric immunoassay was performed using a cell proliferation enzyme-linked immunoassay (ELISA) and a BrDU-colorimetric kit (Roche Diagnostics).

The uterine leiomyoma cells were divided into 1 × 10 ⁴ cells per well of a 96-well plate, cultured for 24 hours, treated with 100 and 200 nM of flavopiridol, and the analytical method was performed according to the manufacturer's instructions. The analytical color was measured at 490 nm.

In order to confirm whether the reduction of viable cell number is due to the proliferation inhibition, the expression level of BrDU incorporated into the DNA through the colorimetric immunoassay was confirmed in the experimental group treated with flavopiridol and the control group not treated.

As a result, as shown in FIG. 2, it was confirmed that the DNA synthesis was reduced in the experimental group treated with the flavopiride, and the result showed that the cell proliferation was reduced. Especially, the decrease of DNA synthesis in the experimental group treated with 200 nM flavopiridol was the greatest, and the decrease of cell viability and cell growth was also confirmed by the decrease of DNA synthesis.

< Example  5> Flavopyridone  Cell cycle changes

To confirm the distribution of the cell cycle, the uterine myoma cells were treated with 100 or 200 nM Flavopyridol for 24 hours, and the cells were harvested, washed with PBS and fixed with 70% ethanol cooled.

The fixed cells were suspended in PBS containing 0.1% RNase A and 50 μg / mL propylidium iodide.

DNA fluorescence was measured using a flow cytometer (FACS Calibur ^™ , Becton Dickinson, Franklin Lakes, NJ, USA) and the percentage of cell cycle steps was determined using ModFit LT ^™ software (Becton Dickinson, USA) Respectively.

As a result of the flow cytometry analysis conducted by the above method in order to confirm the effect of the Flavopyridone on the cell cycle, it was confirmed that the proportion of the uterine myoma cells in the G1 cycle state was increased in a dose dependent manner of Flavopyridone , And 100 nM and 200 nM, respectively, and 43% and 54%, respectively.

In addition, the percentage of cells in the G2 cycle increased to 18% and 22%, while the percentage of cells in the S cycle decreased to 19% and 14%, respectively.

< Example  6> Flavopyridone  Of cell-cycle regulatory proteins in uterine myoma cells

The uterine leiomyoma cells were treated with 100 or 200 nM Flavopyridol for 24 hours and then incubated with 1 × RIPA buffer (Thermo Scientific, Rockford, IL, USA) containing EDTA (Thermo Scientific, USA) and protease and phosphatase inhibitor ) To collect cells and perform Western blot analysis.

Protein concentration was quantified according to the manufacturer's protocol using protein analysis reagent (Bio-Rad, Hercules, Calif., USA), and a portion of the quantified protein was extracted and analyzed on a 8-15% sodium dodecyl sulfate- After electrophoresis, the cells were transferred to a nitrocellulose membrane using a Semi-Dry Transfer Cell (Bio-Rad, USA).

CDK2, CDK4, and retinoblastoma (Rb) (Santa Cruz Biotechnology, USA) were added to the transfer membrane after blocking the protein-transferred membrane with a Tris-buffered saline (TBS) Santa Cruz, CA, USA), p21 ^{cip / waf1} , p27 ^kip1 (BD Pharmingen, Franklin Lakes, NJ, USA), cyclin A (Abcam, Cambridge, Mass., USA) and β-actin (Santa Cruz Biotechnology, USA) and incubated with secondary antibody conjugated with horseradish peroxidase Santa Cruz Biotechnology, USA) and visualized protein bands through a chemiluminescence (ECL) system (Thermo Scientific, USA).

The density of each band was analyzed using a Chemi-doc XRS imaging system (Bio-Rad, USA) and the results were expressed as% control.

Western blot results ^{showed that} the expression of p21 ^{cip / wafl} and p27 ^kip1 was significantly increased while that of CDK2, CDK4, Rb and Cyclin A decreased.

These results indicate that ^{Flavopyridone} reduced the expression of CDK2, CDK4, and Cyclin A and increased the expression of p21 ^{cip / wafl} and p27 ^kip1 , leading to an increase in the G1 phase of uterine leiomyoma cells.

< Example  7> Animal preparation

8-week-old female NSG (NOD-scid IL2Rgamma ^null ) mice were purchased from Jackson Laboratory (USA) and 0.05 mg of 17β-estradiol was added to the sustained-release 17β-estra One aliquot of the diol pellet (Innovative Research of America, Sarasota, Fla., USA) was injected into the mouse to allow 0.05 mg 17β-estradiol to remain in the mice for 60 days.

The 17β-estradiol pellet was injected 2 days before transplantation and the animals were kept in micro-isolator cages separately in sterile environmental facilities and maintained at 12 ° C / .

All breeding materials were sterilized prior to use and the animals were fed diets and water in the laboratory.

< Example  8> Establishment of human mammary gland xenotransplantation animal model

To compare anatomical, physiological and histopathologic changes, we established a traditional transplant animal model and a cell transplant animal model of the new method.

NSG mice were anesthetized by injecting 30 μL of a mixture of ketamine 80 mg / mL and xylazin 20 mg / mL into the abdominal cavity to transplant human myoma uterine tissue or cells into the animal.

Three days after ovariectomy, a small wound of 3 mm was formed on the right side skin of each animal and a mixture of estrogen and progesterone was administered to transplant human myoma tissue. The tissue obtained from the patient of uterine leiomyoma of Example 1 was soaked in Matrigel (Becton Dickinson, USA), inserted into the subcutaneous tissue, and the wound site was closed with suture.

Another method is to transplant human myomaoma cells into NSG mice by gently mixing 1 × 10 ⁷ primary cultured human uterine myoma cells with 0.5 mL Martrigel (Becton Dickinson, USA) as in Example 2 The cells warmed for 5 minutes were injected under the skin of female NSG mice using a 1 cc syringe.

The transplantation of the two methods was completed within 30 minutes and the mice and lesions formed were observed closely once a week.

< Example  9> Histological evaluation of human mammary gland xenograft model

Eight weeks after transplantation, the tissues and animal models of xenograft xenografts were euthanized and pathological evaluations were performed. Tissue was collected and examined for aggregate samples of tissue morphology, and the animal model did not show complications related to anesthesia or surgery.

Tissue collected from the animal model was fixed with formalin and embedded in paraffin to perform hematoxylin and eosin (H & E) histological evaluation.

In addition, immunohistochemistry using anti-alpha smooth muscle actin (αSMA; Dako, USA) and desmin (Abcam, USA) was performed to evaluate typical histologic features of leiomyomata. Anti-ER alpha and anti-PR antibodies (Dako, Carpinteria, CA, USA) were used to evaluate the expression of estrogen receptor (ER) and progesterone receptor (PR).

As a result of the above histological analysis, as shown in FIG. 5, it was confirmed that the H & E stained cell transplantation model clearly distinguished the proliferation of blood vessels of various sizes similar to the tissue graft model.

In addition, αSMA expression was analyzed by specific markers of smooth muscle, and αSMA and desmin expression levels of both transplantation models were compared. As a result, αSMA and desmin protein were strongly positive in both uterine myoma.

However, the tumor was a bundle of vesicular contraction cells of the retina, whereas the uterine leiomyoma cells had elongated and elliptical nucleus and acid cytoplasm, and the abundant extracellular matrix and broad cell blood flow were also confirmed.

As a result of the immunohistochemical analysis using the ER and PR antibodies, the ER and PR protein levels of the two-graft model were similar to those of the cell proliferation.

< Example  10> In xenografted uterine myoma tumors Flavopyridone  Check the effect

To assess the effect of flavopiridol, the volume of the tumor was measured using a digital vernier caliper by hand twice a week, and the volume of the tumor measured (minimum diameter ² x maximum diameter) / 2 Respectively.

The size of the tumor is approximately 30 to 45 mm ³ , The mice were randomly divided into a control group and a flavopiridol test group (n = 5).

In the control group, physiological saline containing 0.1% DMSO was administered to the experimental mice at 5.0 or 7.5 mg / kg of flavopiride for 3 weeks for 5 days in the abdominal cavity, respectively.

The tumor growth was then monitored using a digital vernier caliper for 2 weeks without drug administration and the percentage of tumor volume was calculated at 100% tumor size at the beginning of the administration of flavopiridol.

As a result, referring to FIG. 6A, the tumor growth of the experimental group treated with Flavopyridol after 3 weeks of Flavopyridone treatment was significantly reduced as compared with that of the control group. As a result of analysis of tumor size after completion of Flavopyridone administration Also in Fig. 6C it was found that the volume of the tumor showed a significant decrease.

From the above results, it was confirmed that flavopyridone has an effect of inhibiting the growth of xenotransplantation human uterine myoma.

Hereinafter, formulation examples of the pharmaceutical composition of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically explained.

&Lt; Formulation Example 1 > Preparation of injection

10 mg of flavopyridone, 3.0 mg of sodium metabisulfite, 0.8 mg of methylparaben, 0.1 mg of propylparaben and an appropriate amount of sterilized distilled water for injection were mixed, and the mixture was adjusted to a final volume of 2 ml by a conventional method. Ampicillin and sterilized to prepare an injection.

&Lt; Formulation Example 2 > Preparation of tablet

10 mg of flavopiridol, 100 mg of lactose, 100 mg of starch and an appropriate amount of magnesium stearate were mixed and tableted according to a conventional tablet preparation method.

< Formulation example  3> Preparation of capsules

10 mg of flavopiridol, 50 mg of lactose, 50 mg of starch, 2 mg of talc, and magnesium stearate were mixed and filled in gelatin capsules according to a conventional capsule preparation method to prepare capsules.

Hereinafter, a preparation example of health food using the flavopiridol according to the present invention will be described, but it is not intended to limit the present invention but to describe it specifically.

< Manufacturing example  1> Manufacture of health food

Flavopyridol 10 mg, Vitamin A acetate 70 ㎍, vitamin E 1.0 mg, vitamin B 1 0.13 mg, vitamin B 2 0.15 mg, vitamin B 6 0.5 mg, vitamin B 12 0.2 ㎍, vitamin C 10 mg (1 ug of biotin, 1.7 ㎎ of nicotinic acid amide, 50 ㎍ of folic acid, and 0.5 mg of calcium pantothenate), an appropriate amount of mineral mixture (1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of potassium phosphate, Calcium phosphate (55 mg), potassium citrate (90 mg), calcium carbonate (100 mg) and magnesium chloride (24.8 mg) were mixed to prepare a granule, and a health food was prepared according to a conventional method.

< Manufacturing example  2> Health drink  Produce

10 mg of flavopiridol, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of a plum concentrate, 1 g of taurine and purified water were added to make a total of 900 ml, and the above components were mixed according to a conventional health drink manufacturing method, After stirring and heating at 85 DEG C for about 1 hour, the solution thus prepared was filtered and sterilized in a sterilized 2 L container, followed by refrigerated storage.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (5)

delete delete Wherein the flavopiridol is selectively inhibited cell cycle progression of uterine leiomyoma cells to inhibit cell proliferation and induce apoptosis. Composition. [Claim 5] The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition comprises 0.01 to 10 parts by weight of favopyridone in a total amount of 100 parts by weight. Wherein Flavopyridone is selected from the group consisting of Flavopyridone, Flavopyridone, and Flavopyridone, wherein the Flavopyridone selectively inhibits cell cycle progression of uterine leiomyoma cells to inhibit cell proliferation and induce apoptosis. Food composition.

Images