Composition for prevention, improvement, or treatment of poor ovarian responders caused by aging, comprising butylphthalide or acceptable salt thereof as active ingredient

Abstract

The present invention relates to a composition for prevention, improvement, or treatment of poor ovarian responders caused by aging, comprising butylphthalide or an acceptable salt thereof as an active ingredient, for which a statistically significant increase in the number and quality of mature oocytes was confirmed in 38- to 40-week-old mice (40 years old or older in humans) fed butylphthalide. Not only does the number of mature oocytes produced in the ovaries decrease in women as a result of aging, but also healthy mature oocytes (MII oocytes) decrease as a result thereof, thereby gradually reducing the possibility of pregnancy, but the active ingredient of the present invention may increase the number and quality of mature oocytes and thus has the effect of increasing the chances of pregnancy for older women. In addition, the present invention may inhibit a decrease in the number of primordial follicles caused by aging and thus has the effect of enabling the inhibition of diminished ovarian reserve caused by aging.

Description

Composition for preventing, improving or treating ovarian hyporesponsiveness due to aging, comprising butylphthalide or an acceptable salt thereof as an active ingredient

[0001] The present invention relates to a composition for preventing, improving or treating ovarian hyporesponsiveness due to aging, comprising butylphthalide or an acceptable salt thereof as an active ingredient.

[0002] This work was supported by the 'Development of Application Technology for Herbal Medicine to Solve Industrial Demands' project (Project No. 1711195904, KSN2022330), a basic project of the Korea Institute of Oriental Medicine under (or affiliated with) the Ministry of Science and ICT, and the 'Establishment of a Systematic Oriental Medicine Ecosystem through Digital Conversion of Oriental Medicine Pharmacology' (Project No. 1711195897, KSN1731122), a major project of the Korea Institute of Oriental Medicine under (or affiliated with) the Ministry of Science and ICT.

[0003] Infertility is defined as the inability of a couple to conceive within one year despite regular sexual intercourse without using contraception. Primary infertility refers to a couple who have never previously experienced pregnancy, while secondary infertility refers to a couple who have previously conceived, even if they did not deliver. Infertility can occur if there is an abnormality at any stage throughout the entire process, from the development of male or female gametes (sperm or egg), to fertilization, development of the fertilized embryo, or implantation of the embryo in the uterus.

[0004] Recently, the rate of pregnancy at older ages has been increasing due to factors such as the rising average age of first marriage and the rise in remarriage. Pregnancy and childbirth after age 35 are medically referred to as "old-age pregnancy" or "old-age childbirth." As biological age increases, ovarian function declines, which can lead to infertility. In particular, even if a woman ovulates around age 40, the likelihood of ovulating a healthy egg drops sharply, making pregnancy difficult. Therefore, even if ovulation occurs, it is crucial to ensure that the egg develops normally and produces healthy eggs capable of conceiving.

[0005] If a woman is over 40 years old, has one or more risk factors for ovarian dysfunction, has had three or fewer eggs retrieved during a previous IVF, or has abnormal findings in an ovarian function evaluation, she is considered a poor ovarian responder. In this group, not only does ovulation not occur properly, but there are also many cases where problems occur with the growth of healthy mature eggs.

[0006] Meanwhile, butylphthalide has the effect of improving the damage of central nervous system function in patients with acute ischemic stroke, and can promote the recovery of patients' function, and is mainly converted into two metabolites, 3-3-(3'-hydroxy)butylphthalide and 3-hydroxy-3-butylphthalide, in vivo.

[0007] As for butylphthalide-related technologies, Korean Patent No. 0860703 discloses the use of L-butylphthalide in the manufacture of a drug for preventing and treating cerebral infarction, and Korean Patent No. 1659596 discloses a butylphthalide derivative and a method for producing the same and the use thereof. However, there is no disclosure regarding a composition for preventing, improving, or treating ovarian hyporesponsiveness due to aging, which comprises butylphthalide or an acceptable salt thereof as an active ingredient.

[0008] The present invention was derived from the above-mentioned needs, and the present invention provides a composition for preventing, improving or treating ovarian hyporesponsiveness due to aging, which comprises butylphthalide or an acceptable salt thereof as an active ingredient, and completed the present invention by confirming that the number and quality of mature eggs statistically significantly increased in 38-40 week-old mice (40 years or older in humans) fed butylphthalide.

[0009] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0010] In addition, the present invention provides a health functional food composition for preventing or improving ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0011] In addition, the present invention provides a veterinary composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0012] The present invention relates to a composition for preventing, improving or treating ovarian hyporesponsiveness due to aging, comprising butylphthalide or an acceptable salt thereof as an active ingredient. The butylphthalide or an acceptable salt thereof, which is an active ingredient of the present invention, can improve the number and quality of mature eggs, and thus has the effect of increasing the possibility of pregnancy in elderly women.

[0013] Figure 1 shows the results of confirming body weight (A) and ovarian weight (B) according to butylphthalide administration. YC is a group that was orally administered distilled water for 4 weeks to 8-10 week-old mice, OC is a group that was orally administered distilled water for 4 weeks to 38-40 week-old mice, and OC+butylphthalide (OC+BP) is a group that was administered the butylphthalide of the present invention for 4 weeks to 38-40 week-old mice. ** indicates that the body weight of the OC group was statistically significantly increased compared to the YC group, p<0.01, **** indicates that the ovarian weight of the OC group was statistically significantly decreased compared to the YC group, p<0.0001, and # indicates that the ovarian weight of the OC+BP group was statistically significantly increased compared to the OC group, p<0.05.

[0014] Figure 2 shows the results of confirming the number of follicles by maturation stage according to butylphthalide administration. (A) Primordial follicles, (B) Primary follicles, (C) Secondary follicles, (D) Antral follicles, (E) Total follicles. **, **** indicate a statistically significant decrease in the number of follicles by maturation stage in the OC group compared to the YC group; ** indicates p<0.01, and **** indicates p<0.0001. # indicates a statistically significant increase in the number of primordial follicles in the OC+BP group compared to the OC group; p<0.05.

[0015] Figure 3 shows the results of confirming the number and quality of ovulated eggs according to butylphthalide administration. (A) represents the total number of ovulated eggs, (B) represents the number of mature eggs, and (C) represents the number of mature eggs with normal spindle alignment. **, *** indicate that the total number of ovulated eggs, the number of mature eggs, or the number of mature eggs with normal spindle alignment in the OC group was statistically significantly decreased compared to the YC group. ** indicates p<0.01, and *** indicates p<0.001. # indicates that the number of ovulated mature eggs or the number of mature eggs with normal spindle alignment in the OC+BP group was statistically significantly increased compared to the OC group. p<0.05.

[0016] The present invention relates to a pharmaceutical composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0017] The above butylphthalide is a compound represented by the following chemical formula 1.

[0018]

[0019] The above butylphthalide or an acceptable salt thereof is characterized by inhibiting the decrease in the number of primordial follicles due to aging and increasing ovulation of mature eggs with the potential for fertilization.

[0020] The salts include common acid addition salts, for example, salts derived from inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, and hydrobromic acid, and salts derived from organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, succinic acid, benzoic acid, citric acid, maleic acid, malonic acid, malic acid, tartaric acid, gluconic acid, lactic acid, gestic acid, fumaric acid, lactobionic acid, salicylic acid, phthalic acid, embonic acid, aspartic acid, glutamic acid, and acetylsalicylic acid. The salts also include salts derived from amino acids such as glycine, alanine, valine, isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine, arginine, tyrosine, and proline. Additionally, the salt includes salts of sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, and toluenesulfonic acid.

[0021] In the present invention, a poor ovarian responder is defined as a woman who is 40 years of age or older, has one or more risk factors for ovarian dysfunction, has three or fewer eggs retrieved during a previous superovulation in vitro fertilization procedure, or has abnormal findings observed in an ovarian function evaluation, when two or more of the following apply.

[0022] The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above-mentioned effective ingredient, and may be in various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, etc., and these solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, and aerosols. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers, and suppositories. Non-aqueous solvents and suspending agents can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerol gelatin. For parenteral administration, it is preferable to select a method such as topical application to the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, epidural, or intracerebrovascular injection.

[0023] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment. The level of the effective amount may be determined based on factors including the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. It is important to take all of the above factors into consideration and administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0024] The dosage of the composition of the present invention varies depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity. The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

[0025] In addition, the present invention relates to a health functional food composition for preventing or improving ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0026] The above composition is preferably manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0027] When the health functional food composition of the present invention is used as a food additive, the butyl phthalide may be added as is or used together with other foods or food ingredients, and may be used appropriately according to a conventional method. The amount of the active ingredient mixed may be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may also be used in an amount exceeding the above range.

[0028] There are no specific restrictions on the types of the above foods. Examples of foods to which butylphthalide can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense.

[0029] When the composition of the present invention is used as a health beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, a natural sweetener such as thaumarin or stevia extract, or a synthetic sweetener such as saccharin or aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 g of the composition of the present invention. The composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in mixtures. The proportion of these additives is not particularly important, but the composition of the present invention is typically selected within the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0030] In addition, the present invention relates to a veterinary composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

[0031] The veterinary composition of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, ethanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil.

[0032] The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using a method well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of a powder, granule, tablet, capsule, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, suppository, sterile injection solution, sterile external preparation, etc.

[0033] The effective amount of the veterinary composition according to the present invention can be appropriately selected depending on the individual animal. This may be determined based on factors including the severity of the disease or condition, the individual's age, weight, health status, or sex, sensitivity to the active ingredient of the present invention, the route of administration, the duration of administration, other compositions combined with or used concurrently with the composition, and other factors well known in the fields of physiology and veterinary medicine.

[0034]

[0035] Hereinafter, the present invention will be described in more detail using examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0036]

[0037] Example 1. Confirmation of body weight and ovarian weight according to butylphthalide administration

[0038] YC (Young control): Distilled water (DW) was orally administered 20 times over 4 weeks to 8-10 week-old BALB / c mice (n=6).

[0039] OC (Old control): Distilled water (DW) was orally administered 20 times over 4 weeks to 38-40 week-old BALB / c mice (n=6).

[0040] OC + BP (Old Control + butylphthalide): BALB / c mice (n=6) aged 38 to 40 weeks were orally administered 5 mg / kg / day of butylphthalide 20 times for 4 weeks.

[0041] Body weight was measured at one-week intervals, and after oral administration, 5 IU of PMSG (pregnant mare's serum gonadotropin) was injected intraperitoneally. Two days after PMSG injection, 5 IU of hCG was injected intraperitoneally, and the next day, autopsy was performed to measure ovarian weight.

[0042] As a result, as disclosed in Fig. 1, the body weight of 38-40 week old mice (OC) increased compared to 8-10 week old mice (YC), and in contrast, the body weight of the experimental group (OC+BP) orally administered butylphthalide showed almost no change.

[0043] Meanwhile, the ovarian weight of 38-40 week-old mice (OC) was significantly reduced compared to 8-10 week-old mice (YC), and the ovarian weight of the experimental group (OC+BP) orally administered butylphthalide was significantly increased compared to 38-40 week-old mice (OC).

[0044]

[0045] Example 2. Confirmation of the number of follicles at each stage of follicular maturation according to butylphthalide administration

[0046] After measuring the weight of the oocytes in Example 1, the ovarian tissue was extracted and the distribution of follicle growth stages within the tissue was evaluated. The ovarian tissue was fixed in 4% paraformaldehyde fixative, embedded in paraffin, and then the entire tissue was serially sectioned at 5 μm thickness, and every fifth section was stained with H&E and examined under a microscope. The follicle growth stages in the sections were classified and evaluated as primordial follicles, primary follicles, secondary follicles, and antral follicles.

[0047] As a result, as disclosed in Fig. 2, the number of follicles was significantly reduced in all stages in OC compared to YC, and the number of follicles increased in OC+BP administered with butylphthalide of the present invention compared to OC, and in particular, it was confirmed that butylphthalide, the effective ingredient of the present invention, suppresses the decrease in ovarian reserve in mice due to aging through the increase in primordial follicles (Fig. 2(A)).

[0048]

[0049] Example 3. Confirmation of the number and quality of ovulated eggs following administration of butylphthalide.

[0050] YC (Young control): Distilled water was orally administered 20 times over 4 weeks to 8-10 week-old BALB / c mice.

[0051] OC (Old control): Distilled water was orally administered 20 times over 4 weeks to 38-40 week-old BALB / c mice.

[0052] OC+BP: BALB / c mice aged 38 to 40 weeks were orally administered 5 mg / kg / day of butylphthalide 20 times for 4 weeks.

[0053] After oral administration, 5 IU of PMSG (pregnant mare's serum gonadotropin) was injected intraperitoneally, and on the second day, hCG was injected to induce ovulation, and the ovulated oocytes were collected and evaluated for number and quality.

[0054] As a result, as disclosed in Fig. 3, although there was no difference in the total number of ovulated oocytes, not only did the number of mature oocytes (MⅡ) increase in the OC+BP group, but the number of MⅡ oocytes with normal spindle alignment, which have the highest probability of being fertilized, also increased statistically significantly.

Claims

1. A pharmaceutical composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

2. A pharmaceutical composition for preventing or treating ovarian hyporesponsiveness due to aging, characterized in that in paragraph 1, the butylphthalide or an acceptable salt thereof suppresses a decrease in the number of primordial follicles due to aging.

3. A pharmaceutical composition for preventing or treating ovarian hyporesponsiveness due to aging, characterized in that the butylphthalide or an acceptable salt thereof in paragraph 1 increases ovulation of mature oocytes with a possibility of fertilization.

4. A pharmaceutical composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, characterized in that in paragraph 1, in addition to the effective ingredient, it further comprises a pharmaceutically acceptable carrier, excipient or diluent.

5. A pharmaceutical composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, characterized in that the composition in the first paragraph is manufactured in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols.

6. A health functional food composition containing butylphthalide or an acceptable salt thereof as an effective ingredient for preventing or improving ovarian hyporesponsiveness due to aging.

7. A health functional food composition for preventing or improving ovarian hyporesponsiveness due to aging, characterized in that the composition in claim 6 is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candy, syrup, and beverage.

8. A veterinary composition for the prevention or treatment of ovarian hyporesponsiveness due to aging, containing butylphthalide or an acceptable salt thereof as an active ingredient.

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