JP2023156553A - Ovarian function activators, anti-aging agents, pharmaceuticals, cosmetics, and foods or beverages - Google Patents

Abstract

To provide an ovarian function activator that can maintain or improve ovarian function by enhancing the homing effect to the ovary.SOLUTION: An ovarian function activator comprises: at least one of the group consisting of isoflavone, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B1, vitamin B2, and vitamin B6; and stem cells.SELECTED DRAWING: None

Description

The present invention relates to ovarian function activators, anti-aging agents, pharmaceuticals, cosmetics, and foods or beverages.

When a woman is born, all of her eggs are already present in her ovaries, but as she ages, the number of eggs decreases and their quality declines. Nowadays, the age at which women get married is increasing year by year, and even if there is no obvious cause of infertility, an increasing number of women have difficulty conceiving due to age-related declines in ovarian function, i.e., a decline in the number and quality of eggs in the ovaries. It is also said that there are Furthermore, a decline in ovarian function is said to affect the appearance of women, such as the luster of their skin.

Although it is difficult to newly generate oocytes and oogonia stem cells (OSC), it is possible to maintain or improve ovarian function by stimulating the differentiation of stem cells introduced from outside the body and increasing the homing effect to the ovary. If this can be achieved, it will lead to the resolution of various problems faced by women.
In other words, maintaining and improving ovarian function is an essential factor for anti-aging, maintaining or restoring women's beauty and health.

As for the homing effect on germ cells, for example, a testicular function improving agent is proposed in Patent Document 1. Patent Document 1 describes that a testicular function improving agent circulates within the testis, and when it finds damaged tissue (including stem cells), the homing effect activates the tissue (stem cells) themselves and repairs and regenerates them. There is.

Patent No. 6830286

An object of the present invention is to provide an ovarian function activator, an anti-aging agent, a pharmaceutical, a cosmetic, and a food or beverage that can maintain or improve ovarian function by enhancing the homing effect to the ovary. .

As a result of extensive studies, the present inventors have discovered that the above object can be achieved by blending predetermined components, and have completed the present invention.

That is, according to the present invention,
(1) An ovarian function activator comprising at least one member of the group consisting of isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ , and stem cells;
(2) an anti-aging agent containing the ovarian function activator according to (1);
(3) Pharmaceutical products containing the ovarian function activator described in (1),
(4) Cosmetics containing the ovarian function activator according to (1) (5) A food or drink containing the ovarian function activator according to (1) is provided.

According to the present invention, there are provided ovarian function activators, anti-aging agents, pharmaceuticals, cosmetics, and foods or beverages that can maintain or improve ovarian function by enhancing the homing effect to the ovaries.

It is a figure showing the measurement result of the amount of urinary estrogen in an example. FIG. 3 is a diagram showing the measurement results of the amount of AMH in blood in Examples. FIG. 3 is a diagram showing measurement results of skin moisture content and hyaluronic acid content in Examples.

Hereinafter, the ovarian function activating agent of the present invention will be explained. The ovarian function activator of the present invention contains a group consisting of isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B1, vitamin B2, and vitamin B6 (hereinafter sometimes referred to as "compounded ingredients"). and stem cells.

(Ingredients)
The ovarian function activator of the present invention comprises a group consisting of isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ (hereinafter sometimes referred to as "compounded ingredients"). .).

As the isoflavone, 3-phenylchromone can be preferably used, but isoflavones, which are flavonoids having 3-phenylchromone as a basic skeleton, may be used in part or in whole.

Equol specifically refers to 7-hydroxy-3-(4'-hydroxyphenyl)-chroman. Here, equol is a substance produced by metabolizing daidzein, which belongs to isoflavones, by intestinal bacteria, digestive enzymes, etc. Equol used in the present invention is a synthetic substance obtained by chemically reducing daidzein. Equol may be used, or a metabolite obtained by metabolizing a raw material containing daidzeins by equol-producing bacteria, which is a type of intestinal bacteria, may be used. Note that daidzeins include daidzein, dihydrodaidzein, daidzein glycosides, and the like.
Glutathione is a tripeptide consisting of glutamic acid, cysteine, and glycine bonded in this order.
L-carnitine is a substance synthesized from lysine and methionine.

bFGF (basic fibroblast growth factor) is a proliferation (growth) factor that exhibits pleiotropic actions. In addition, basic fibroblast growth factors include basic fibroblast growth factor 7 (FGF-7), also known as keratinocyte growth factor KGF, KGF-2, and basic fibroblast growth factor 10 (FGF-2). 10) may be mentioned, but in the present invention, it is preferable to use bFGF (or also referred to as FGF2).

Vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ are vitamins, and may be synthesized or extracted from natural products.

In the following, the product containing all isoflavones, equol, glutathione, L-carnitine, bFGF (basic fibroblast growth factor), vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ will be referred to as "Stem-X". However, the blending ratio of each component constituting Stem-X is not particularly limited.

(stem cells)
Stem cells used in the ovarian function activating agent of the present invention are not particularly limited, but include mesenchymal stem cells, hematopoietic stem cells, neural stem cells, epidermal stem cells, embryonic stem (ES) cells, embryonic reproductive (EG) cells, and ovarian-derived stem cells. Stem cells, all other tissue stem cells, etc. can be used. Among these, ovary-derived stem cells can be preferably used. Furthermore, when using stem cells, it is preferable to use them in a state containing exosomes.

Further, the stem cells may be of human origin (autologous cells) or of xenogeneic origin (allogeneic cells). Species of stem cells derived from different species include cows, horses, pigs, dogs, cats, mice, rats, sheep, and the like. In addition, when using autologous cells, the cells may be one's own or cells from another person may be used.

(ovarian function activator)
The ovarian function activating agent of the present invention contains the above-mentioned ingredients and stem cells. Further, the blending ratio of the ingredients to the stem cells is not particularly limited, but is preferably 5:95 to 95:5 on a weight basis.

The ovarian function activator of the present invention may be used as is by combining the above-mentioned ingredients and stem cells at a certain ratio, or by adding the above-mentioned ingredients to the stem cells and adjusting the mixture under predetermined conditions. It may also be used after culturing.

The ovarian function activator of the present invention comprises a certain amount of several combination reagents (at least one of the group consisting of isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ ). ) to the mesenchymal stem cell culture medium, the administered stem cells or stem cells originally existing in the body are induced to become oocytes and oogonia stem cells (OSC)-like cells, bringing other stem cells with them. It gathers in the ovaries and promotes regeneration, resulting in improved ovarian function.

Specifically, an increase in the number of eggs in the ovary and an increase in the amount of secreted hormones (estrogen, etc.) are indicators of increased ovarian function, and as a ripple effect, skin aging is inhibited, especially to suppress sagging. You can expect good results.

Another indicator of ovarian function is the age equivalent of the number of eggs remaining in the ovary, which is called ``ovarian age.'' Ovarian age is calculated based on the measurement results of AMH (Anti-Mullerian Hormone), a hormone released by cells surrounding the egg. In addition, skin age is influenced by moisture content, skin viscoelasticity, skin brightness, and wrinkles resulting from destruction of collagen and elastin due to increased gelatinase.

Although it is difficult to newly generate oocytes and oogonia stem cells (OSCs), according to the present invention, stem cells introduced from outside the body are stimulated to induce differentiation, thereby increasing the homing effect to the ovary. Functions can be maintained or improved.

(Application)
The ovarian function activator of the present invention is intended to maintain and improve beauty and health, and is used for purposes of maintaining and improving beauty and health, and for anti-aging purposes. be able to. In other words, it can be used for the above-mentioned purposes rather than for the treatment of diseases peculiar to women, so it can be used not only by all women, but also by men, considering its use for younger-looking skin. be.

Furthermore, since the ovarian function activator of the present invention has anti-aging effects, it can also be incorporated into pharmaceuticals intended for these purposes. As a pharmaceutical, it can be used as either a preventive drug or a therapeutic drug.

When incorporated into pharmaceuticals, the component combination product may be used alone, or may be mixed with additives that are generally acceptable in pharmaceutical formulations to form a formulation. In addition, the dosage forms include oral dosage forms such as tablets, granules, capsules, pills, powders, solutions, suspensions, emulsions, syrups, elixirs, and extracts, or injections, Examples include dosage forms using parenteral agents such as liquids, suppositories, ointments, patches, poultices, and lotions; however, there are no particular limitations, and the dosage form should be selected as appropriate depending on the purpose of treatment or prevention. Can be done.

Furthermore, in the case of tablets, granules, pills, capsules, and powders, additives such as excipients, binders, disintegrants, and lubricants can be included. Examples of excipients include starch, carboxymethylcellulose, sucrose, dextrin, cornstarch, and the like.

As a binder, crystalline cellulose, crystalline cellulose/carmellose sodium, methylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carmellose sodium, ethylcellulose, Carboxymethylethyl cellulose, hydroxyethyl cellulose, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, pullulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer E, aminoalkyl methacrylate copolymer RS , methacrylic acid copolymer L, methacrylic acid copolymer, polyvinyl acetal diethylaminoacetate, polyvinyl alcohol, gum arabic, gum arabic powder, agar, gelatin, white shellac, tragacanth, refined white sugar, and macrogol.

Disintegrants include crystalline cellulose, methylcellulose, low-substituted hydroxypropylcellulose, carmellose, carmellose calcium, carmellose sodium, croscarmellose sodium, wheat starch, rice starch, corn starch, potato starch, partially pregelatinized starch, hydroxy Examples include propyl starch, sodium carboxymethyl starch, and tragacanth.

As lubricants, wheat starch, rice starch, corn starch, stearic acid, calcium stearate, magnesium stearate, hydrated silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, dry aluminum hydroxide gel, talc, aluminum metasilicate Examples include magnesium acid, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, sucrose fatty acid ester, waxes, hydrogenated vegetable oil, and polyethylene glycol.

In addition, in the case of solutions, syrups, suspensions, emulsions, and elixirs, in addition to commonly used inert diluents such as water and vegetable oil, coloring agents, flavoring agents, flavoring agents, etc. are added. It may also be included as an agent.

Furthermore, in the case of injections, additives such as suspensions, emulsions, and solubilizers for use can be included. In the case of ointments and suppositories, additives such as fat, fatty oil, lanolin, petrolatum, paraffin, wax, resin, plastic, base, glycols, higher alcohol, water, emulsifiers, suspending agents, etc. It can be contained as. In the case of poultices, glycerin, water, water-soluble polymers, water-absorbing polymers, etc. can be contained as additives. Further, in the case of a lotion, a solvent, an emulsifier, a suspending agent, etc. can be included as additives.

Moreover, the ovarian function activator of the present invention can also be blended into cosmetics. Cosmetics include lotions, milky lotions, face washes, cleansers, serums, creams, foundations, eyebrows, mascara, eye shadows, eyeliners, lipsticks, glosses, cheeks, white powder, nail polish, and the like. Furthermore, the cosmetics may be in the form of liquid, cream, solid, stick, powder, or the like.

Furthermore, the ovarian function activator of the present invention may be incorporated into foods, drinks, and the like. Examples of foods include breads, noodles, confectionery, processed meat products, processed seafood products, frozen foods, jellies, ice creams, dairy products, and various seasonings. In addition to general foods, it can also be incorporated into foods for specified health uses, quasi-drugs, health foods, and supplements. Beverages include soft drinks, milk drinks, alcoholic beverages, tea, black tea drinks, coffee, fruit juice drinks, carbonated drinks, mineral waters, fruit and vegetable drinks, and the like.
Furthermore, foods and drinks containing the ovarian function activating agent of the present invention may be in the same form as oral preparations such as tablets, capsules, and syrups.

In addition, when producing foods and beverages containing the ovarian function activator of the present invention, sweeteners, colorants, preservatives, thickeners, Additives such as stabilizers, gelling agents, antioxidants, coloring agents, bleaching agents, emulsifiers, swelling agents, acidulants, brighteners, fragrances, solvents, and oils may be added. Each type of these additives may be used alone, or two or more types may be used in combination.

The proportion of the ovarian function activator of the present invention blended into the above foods and drinks can be adjusted as appropriate depending on the purpose of use, but the proportion of the ingredient combination product blended into the above foods and drinks: The amount is preferably 0.01 to 20% by weight, more preferably 0.01 to 15% by weight, and even more preferably 0.1 to 10% by weight.

The present invention will be explained below with reference to Examples, but the present invention is not limited thereto.
A "Stem-X" solution was prepared by dissolving isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ in ultrapure water or physiological saline.

The "Stem-X" solution and mouse ovary-derived stem cell suspension were administered in 0.5 ml via the tail vein to old mice (ICR; 25 weeks old, female). Similarly, groups to which only stem cells and only solution were administered were also created. Four groups of mice, including mice in the non-administration group, were observed for 3 weeks and 6 weeks under the same breeding conditions. In the 6-week group, after 3 weeks had elapsed, an additional administration similar to the first administration was conducted (second administration). During the observation period, the same chow and drinking water were available ad libitum. Urine was collected from mice in each group after 3 weeks and 6 weeks, and the amount of urinary estrogen (oestrone, 17β-estradiol, estriol) was measured by ELISA method (Estrone; K031-H1, 17β-estradiol; K030-H1, Estriol). ; K064-H1, all measured using Arbor Assays). In addition, after collecting blood from the ophthalmic plexus and jugular vein under Nembutal anesthesia, the amount of AMH (anti-Mullerian hormone) in the blood was measured using the Anti-Mullerian Hormone (AMH) ELISA Kit (Cloud-Clone Corp. Wuhan; CEA228BO). .

Since the estrous cycle of mice is said to last 4 to 5 days, the amount of urinary estrogen was measured at the 3rd week (6wks), the 6th week (6wks), from the 21st to the 25th, and from the 42nd to the 46th, respectively. Measurements were made for 5 consecutive days, and expressed as a percentage, with the value of non-administered mice of the same week taken as 100%. AMH measurements were performed by ophthalmic venous plexus blood sampling on the 21st and 42nd days, and by jugular vein blood sampling on the 25th and 46th days. (The 3-week experimental group and the 6-week experimental group were conducted using different mice.)

In addition, the skin from the back of the mouse was collected on Day 25 from the start of the experiment and at the end of the experiment on Day 45, and the water content was determined using normal pressure heat drying method (MX-50; A&D Co., Ltd.). The amount of hyaluronic acid in the skin was measured using the ELISA method (Hyaluronan Quantikine ELISA Kit; manufactured by R&D systems). The skin for measuring hyaluronic acid was freeze-ground for each individual, and the skin was ground under liquid nitrogen until it became powder. The obtained frozen and crushed skin was defatted in ethanol (99.5%) at 4°C overnight. Next, add PBS(-) containing protease inhibitor (5 mM benzamidine hydrochloride/10 mM EDTA-2Na/0.1 M aminohexanoic acid dissolved in PBS(-), pH 7.4) and incubate at 4°C for 3 to 4 hours. After rotating with a rotator for a time, centrifuging, and removing the supernatant, the precipitate was dissolved in PBS(-) containing a protease inhibitor and used as a sample for ELISA.

The measurement results of the amount of estrogen in the urine are shown in FIG. 1, the results of the measurement of the amount of AMH in the blood are shown in FIG. 2, and the results of the measurement of the amount of skin moisture and hyaluronic acid are shown in FIG. 3, respectively. The experimental groups in FIGS. 1 to 3 are as follows.
Group 1; Physiological saline administration group 2; Mouse ovary-derived stem cell suspension group 3; Stem-X solution administration group 4; Mouse ovary-derived stem cell suspension and Stem-X solution administration group

As shown in Figure 1, the measurement results of urinary estrogen (estrone, 17β-estradiol, estriol) were almost the same at 3 weeks (3wks) and 6 weeks (6wks), and group 2 was more effective than group 1. (Group to which mouse ovary-derived stem cell suspension was administered alone) and Group 3 (Group to which Stem-X solution was administered alone) had higher estrogen levels. Furthermore, the estrogen level was higher in Group 4 (the group to which both the mouse ovary-derived stem cell suspension and Stem-X solution were administered) than in Groups 2 and 3.

As shown in Figure 2, the measurement results for the amount of AMH in serum had the same tendency as that for urinary estrogen, and were higher in group 2 (the group in which the mouse ovary-derived stem cell suspension was administered alone) and group 3 (in which the mouse ovary-derived stem cell suspension was administered alone) than in group 1. The amount of AMH was higher in the group to which Stem-X solution was administered alone. Furthermore, the amount of AMH was higher in Group 4 (the group to which both the mouse ovary-derived stem cell suspension and Stem-X solution were administered) than in Groups 2 and 3. Furthermore, higher results were shown in the 6th week (6wks; 2-time administration group) than in the 3rd week (3wks; 1-time administration). This suggests the advantage of regular administration.

As shown in FIG. 3, both the skin moisture content and the hyaluronic acid content were high in Groups 3 and 4, and this trend was the same at 3 weeks (3wks) and 6 weeks (6wks). By administering the drug twice, the amount of hyaluronic acid in Group 4 was the highest.

The body hair of the mice was apparently smoother in groups 2, 3, and 4 than in group 1 throughout the experimental period. In addition, no differences were observed in mouse body weight or ovary weight between groups. Although it is possible that changes may be observed with repeated administration over a longer period of time, it has been confirmed that internal indicators (urine, blood) have already improved in the positive direction after the two administrations.

Based on these results, this reagent, "Stem-X", when added to a stem cell culture medium outside the body or introduced into the body together with stem cells, has the ability to direct stem cells to the ovaries (homing effect) and cause them to enter the body. It was assumed that ovarian function would be enhanced by attracting pre-existing stem cells or acting directly on the ovaries. Furthermore, it has been suggested that "Stem-X" suppresses skin aging as a side effect of circulation in the body.

Claims (5)

at least one member of the group consisting of isoflavones, equol, glutathione, L-carnitine, bFGF, vitamin C, vitamin B ₁ , vitamin B ₂ and vitamin B ₆ ;
An ovarian function activator containing stem cells. An anti-aging agent containing the ovarian function activator according to claim 1. A pharmaceutical product containing the ovarian function activating agent according to claim 1. A cosmetic product containing the ovarian function activator according to claim 1. A food or drink containing the ovarian function activator according to claim 1.