KR101583777B1 - Novel use of arctiin - Google Patents

Abstract

In the present invention, provided is a novel use of arctiin. According to the present invention, it is possible to effectively treat, ameliorate, and/or prevent endometriosis by using arctiin of the present invention. Further, in the present invention, provided is a pharmaceutical composition for treating or preventing endometriosis, which contains arctiin as an active ingredient, and inhibits growth, migration ability, or adhesiveness of endometriotic cells.

Description

Novel use of arctin < RTI ID = 0.0 >

The present invention relates to new uses of arctin, and more particularly to novel medicinal or food uses of arctin.

Endometriosis is a very common gynecologic disease that affects about 3% to 10% of women in fertility and 25% to 35% of infertile women. It causes abnormally proliferating endometrial tissue outside the uterus (ovary, abdominal cavity, .

Endometriosis is a symptom of various pain associated with dysmenorrhoea, dyspareunia, and invasion sites and has been identified as the cause of infertility in 30-50% of patients.

Until now, there has been no fundamental treatment for endometriosis except surgical removal of the endometrium, and this surgery can limit future pregnancy and recurrence rate is high.

Although endometriosis is not a fatal condition, it is associated with severe discomfort due to severe pain and severe infertility. It should be recognized seriously in the quality of life and pregnancy of women, There is no preventive and remedial method yet.

Therefore, there is an urgent need to develop a new concept of endometriosis prevention and treatment, which is a natural but low side effect, not invasive surgery for women, especially for women of childbearing age.

On the other hand, arctin is known to have a skin whitening effect and the like (Korean Patent No. 10-0855457).

Korean Patent 10-0855457, September 1, 2008, Claims

A problem to be solved by the present invention is to provide a novel use of a compound which is effective for treating, ameliorating or preventing endometriosis.

The object of the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

The present inventors have surprisingly found that arctin is effective for the treatment, improvement and / or prevention of endometriosis, thus completing the present invention.

The present invention provides medicinal and / or food uses for the treatment, improvement and / or prevention of endometriosis of arctin.

The present invention also provides a pharmaceutical composition for treating or preventing endometriosis, which comprises actin as an active ingredient.

(3R, 4R) -4 - [(3,4-dimethoxyphenyl) methyl] -3- [3-methoxy- 4S, 5S, 6R) -3,4,5-trihydroxy-6- (hydroxymethyl) oxan-2-yl] oxyphenyl] methyloxolan-2-one.

Actin may be synthesized or isolated from a natural product containing an actin (e.g., urine, etc.) and may be self-manufactured or commercially available. The arctin may be in the form of a pharmaceutically acceptable salt thereof, or may be in the form of a solvate or a prodrug.

The treatment or prophylaxis may be based on one or more selected from the group consisting of inhibiting growth of endometriotic cells by the active ingredient, inhibiting the adherence of endometriotic cells, or inhibiting the migration of endometriotic cells.

Inhibition of cell growth implies inhibition of cell survival, and inhibition of cell survival is implicated in cell death.

The present invention also provides a food composition for improving or preventing endometriosis, which comprises actin as an active ingredient.

Unless otherwise indicated, the food composition is equally applied to the pharmaceutical composition of the present invention, unless otherwise stated. This " improvement " is included in the " treatment " and means that the condition or symptom is improved.

The food composition may be variously contained in foods containing beverages, and may be in the form of beverages, gums, tea, health functional foods, etc. The health functional foods may be formulated into tablets, capsules and the like. The term "health functional food" as used herein refers to foods prepared (including processing) by using raw materials or ingredients having useful functions in accordance with the Korean Health Functional Foods Act No. 12669, and "functional" Refers to the structure and function of the human body to obtain nutritionally useful effects such as controlling nutrients and physiological functions. The food composition may comprise conventional food additives, and the food additives may be selected from natural compounds such as ketones, chemical compounds such as glycine, sodium citrate, nicotinic acid, cinnamic acid, etc., Additives, L-sodium glutamate preparations, noodle-added alkalies, preservative preparations, tar coloring preparations and the like.

The present invention also provides a method for the treatment or prevention of endometriosis comprising the step of administering actin to a mammal, including a human, in need of treatment with actin, and for the manufacture of an agent for the treatment or prophylaxis of endometriosis of actin to provide. The administered arctin may be an effective amount of arctin.

Unless otherwise indicated, the recitation of the pharmaceutical compositions of the present invention is equally applicable to the methods and uses of the present invention, unless they are contradictory.

The actin or composition may be administered orally or parenterally to a mammal, including a human, and the active ingredient may be formulated together with a pharmaceutically acceptable carrier to formulate and administer. In the case of formulation, diluents or excipients such as fillers, extenders, binders, humectants, disintegrants and surfactants which are usually used can be used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose and And / or gelatin. In addition, lubricants such as magnesium and talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweetening agents, fragrances and / or preservatives, and the like are used in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include injectable solutions, suspensions, emulsions, lyophilisers, nasal cleansers, and suppositories. Injectable solutions, suspensions and emulsions may be prepared by mixing water, non-aqueous or suspensions, and active ingredients. Examples of non-aqueous and suspensions include vegetable oils such as propylene glycol, polyethylene glycol and olive oil, ethyl oleate , And the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and / or gelatin can be used. For parenteral administration, it can be administered by subcutaneous injection, intravenous injection, or intramuscular injection.

Actin, included in the composition of the present invention or used in applications and methods, can be used at a dose of 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, on an adult basis per day. The administration can be administered once or several times a day. However, the scope of the present invention is not limited by the dose and the number of administrations.

The composition of the present invention may contain the active ingredient in an amount of 0.1 to 99.9% by weight based on the total weight of the composition.

The above-mentioned arctin can be formulated by adding a pharmaceutically or pharmacologically acceptable carrier, excipient or diluent. For formulation, Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA et al. Can be referenced.

The present invention can effectively treat, ameliorate and / or prevent endometriosis.

FIG. 1 is a graph showing the results of experiments for the treatment, improvement or prevention of endometriosis caused by inhibition of growth of endometriosis cells of actin.
FIG. 2 is a graph showing the results of an experiment for confirming the treatment, improvement or prevention of endometriosis by inhibiting the endometriosis cell migration of actin.
FIG. 3 is a graph showing the results of experiments for the treatment, improvement or prevention of endometriosis by inhibiting the adherence of endothelial cells of actin.

Hereinafter, the present invention will be described in more detail with reference to Examples and Preparation Examples. However, the following examples and preparative examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

By confirming the effect of arctin on endometriosis cell growth, endometriotic cell migration, and endometriotic cell adhesion which are characteristic of endometriosis, it was confirmed by the following method that arctin has the treatment, improvement or prevention effect of endometriosis.

In the following examples, RPMI 1640 medium, DMEM / F12 medium was obtained from Life Technologies (Grand Island, NY, USA) and cell tracker CMFDA was obtained from Invitrogen (Paisley, UK). Unless otherwise noted, reagents were obtained from Sigma.

Example 1: Treatment, improvement or prevention of endometriosis by inhibiting growth of endometriosis cells of actin

1-1. Arctin preparation

(CAS number: 20362-31-6, product number: ASB-00001977-025, IUPAC: (3R, 4R) -4 - [(3,4-dimethoxyphenyl) methyl] -3- [3- 2-yl) oxyphenyl] methyloxolan-2-one) is the primary analytical standard grade, and the concentration of methoxy-4 - [(2S, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy-6- hydroxymethyloxan- Were obtained from ChromaDex (Irvine, CA 92618 USA).

1-2. Cell preparation

A 12Z cell line, human immortalized endometriotic epithelial cells, developed using active endometriosis tissue of endometriosis women was prepared as a endometriosis model cell (Johann-Wolfgang-Goethe-Universitaet (Germany) Provided by Starzinski-Powitz}. These cells have been shown to be similar in molecular biology to active endometriotic tissues (Banu et al., 2008) and are being used as a new in vitro model for molecular biology studies of endometriosis.

Cells were cultured in RPMI 1640 and DMEM / F12 medium (10% fetal bovine serum, 100 U / ml penicillin G, 100 g / ml streptomycin added; Life Technologies, Grand Island, NY) % Air condition.

1-3. Cell viability assay

Cell viability assays were performed with MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-dipheyltetrazoliumbromide; Sigma-Aldrich). Cells were seeded in 96-well plates at a concentration of 5 × 10 ³ cells per well and cultured for 24 hours. To test the growth inhibitory effect of arctin, each cell was treated with arctin (0, 1, 2, 4, 8 uM) prepared at 1-1 for 4 days. On the day of harvest, 25 ul of MTT solution was added to the medium, and the cells were incubated for 4 hours at 37 ° C. After removal of MTT stained media, cells were lysed in DMSO (50 ul) for 30 min. To determine cell viability, optical density was measured at 540 nm using a microplate spectrometer (SpectraMax; Molecular Devices, Sunnyvale, Calif., USA).

The results are shown in Fig. FIG. 1 shows cell survival according to the treatment concentration of actinine. The x-axis represents the concentration of actin treatment and the y-axis represents the degree of cell survival.

As shown, it can be seen that the arctin-treated group suppresses endometriotic cell growth compared to the untreated group.

These results suggest that arctin inhibits the growth of endometriotic cells.

As a result, it can be seen that arctin can effectively treat, ameliorate or prevent endometriosis.

Example 2: Treatment, improvement or prevention of endometriosis by inhibition of endometriosis cell migration of actin

Because endometriosis is characterized by the migration of endometriotic cells, the transwell migration assay is used to determine whether arctin inhibits the endometriosis by inhibiting endometriotic cell migration. Respectively.

2-1. Arctin preparation

Actin was prepared in the same manner as in 1-1 of Example 1.

2-2. Cell preparation

Cells were prepared in the same manner as in 1-2 of Example 1.

2-3. Transwell migration assay

To investigate the effect of 2-1. Arctin treatment on endometriosis cell line (12Z) (treatment concentration: 0, 5, 25, 50 uM) and 16 and 24 hours after endometriosis, A transwell-migration assay was performed.

Specifically, 12Z cells, which are endometriotic cells, were placed in an upper part of a transwell plate in a 24 well-transwell plate (8 um pore size), and treated with arctin. After 16 hours or 24 hours of actin treatment, cells that migrated to the membrane on the outer surface of the transwell were fixed with methanol and stained with 0.5% crystal violet for 10 minutes. Cells migrating to the outer surface of the above-described transwell were observed under a microscope at a magnification of 200 X and were counted and plotted. The results are shown in Fig. The left graph of FIG. 2 shows the results after 16 hours, and the right graph shows the results after 24 hours. In each graph, the x-axis represents the concentration of actin treatment and the y-axis represents the degree of endometriotic cell migration.

As shown in FIG. 2, it can be seen that arctin suppresses the mobility of endometriotic cells.

As a result, it can be seen that arctin can effectively treat, ameliorate or prevent endometriosis.

Example 3 Treatment, Improvement or Preventive Effect of Actin Endometriosis by Suppressing Adhesion of Endometriosis Cells

Endometriosis is characterized by the attachment and implantation of endometrial cells to various organs around the peritoneum and peritoneum.

To confirm whether arctin inhibits the adherence of endometriotic cells, we tried to confirm whether or not the effect of treatment of endometriosis is manifested.

3-1. Arctin preparation

Actin was prepared in the same manner as in 1-1 of Example 1.

3-2. Cell preparation

Cells were prepared in the same manner as in 1-2 of Example 1.

3-3. Experiment on endometriosis cell adhesion

Endothelial cell line (12Z) was treated with 3-1. Arctin (0, 5, 25, and 50 uM) and the extent of endometrial cell adhesion was checked over time (90 min, 120 min).

Specifically, in order to confirm whether or not the actin inhibits the adherence and implantation of endometriotic cells characteristic of endometriosis in organs around peritoneal peritoneal cavity, Met-5A cells (human peritoneal methothelial cells) Type Culture Collection, ATCC) was inoculated with 10% FBS, 100 U / ml penicillin G and 100 g / ml streptomycin (Life Technologies, Grand Island, NY, USA) and 400 nM hydrocortisone (St. Louis, MO, USA) in a 5% CO ₂ incubator. The Met-5A cells were dispensed into 96-well plates. Cell tracker green 12X cells (2 × 10 ³ cells / well) of the endometriosis cell line labeled with CMFDA were mixed with DMEM / F12 medium containing arctin and then added to previously dispensed Met-5A cells. The cells were incubated in a 5% CO ₂ incubator at 37 ° C for one hour, and then the plates were flipped and washed with a phosphate-buffered solution containing calcium and magnesium. Attached cells were analyzed at a wavelength of 490 nm and the percentage of cells attached was calculated by placing the control at 100%. The results are shown in the graph of FIG. The x-axis of the figure represents the incubation time (min) of arctin, and the y-axis represents the percent attachment of endometriotic cells.

As a result, it can be seen that actin inhibits adherence of endometriotic cells in a concentration-dependent manner.

From these results, it can be seen that arctin inhibits the adherence of endometriotic cells, thereby effectively treating, improving or preventing endometriosis.

As can be seen from Examples 1 to 3 described above, it can be seen that arctin inhibits endometriosis cell growth, endometriotic cell migration, and endometriotic cell adhesion, which are characteristic of endometriosis.

Thus, it can be seen that arctin is effective in the treatment, amelioration or prevention of endometriosis.

≪ Preparation Example 1 > Preparation of pharmaceutical composition

300 mg of arctin, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, prepared in the same manner as in Example 1-1, were filled in gelatin capsules to prepare capsules.

≪ Preparation Example 2 > Preparation of food composition

Water was added to the arctin (4% by weight), liquid fructose (0.5% by weight), oligosaccharide (2% by weight), sugar (2% by weight) and salt (0.5% by weight) prepared in the same manner as in Example 1-1 The remaining amount was adjusted, and homogeneously blended and instant sterilized to prepare a health drink.

Claims (8)

Containing actin as an active ingredient,
A pharmaceutical composition for treating or preventing endometriosis, which inhibits growth of endometriotic cells, migration ability of endometriotic cells, or adherence of endometriotic cells. delete delete delete Containing actin as an active ingredient,
A composition for improving endometriosis, which inhibits growth of endometriosis cells, migration ability of endometriotic cells, or adherence of endometriotic cells. delete delete delete

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