Application of pyrroloquinoline quinone or salt thereof

Abstract

The present invention relates to the use of pyrroloquinoline quinone or a salt thereof in the preparation of an inhibitor or ameliorating agent for female reproductive tissue fibrosis, an ameliorating or prophylactic agent for irregular menstruation, an ameliorating or prophylactic agent for ovarian hyperstimulation syndrome, or an ameliorating agent for abnormal ovulation cycle.

Description

[0001] The present application is a divisional application of Chinese Patent Application No. 202080045230.3, filed on June 22, 2020, entitled “Female Anti-aging Agent”, which claims priority to Japanese Patent Application No. 2019-121736, filed on June 28, 2019. TECHNICAL FIELD

[0002] The present application relates to a female anti-aging agent. BACKGROUND

[0003] For women, as they age, reproductive tissues such as ovaries, follicles, and oocytes gradually age and become difficult to conceive or give birth. For the treatment of infertility in such women who are difficult to conceive, hormone replacement therapy is often used to promote ovulation. However, in addition to the difficulty in developing and expelling uniformly mature follicles, as a side effect, a symptom called ovarian hyperstimulation syndrome (OHSS) occurs, and if this symptom worsens, there is a risk of falling into respiratory failure or circulatory failure. Therefore, for the treatment of infertility, a safe and effective treatment method is being sought (Non-Patent Literature 1).

[0004] In addition, as the process of aging of female reproductive tissues progresses, if the concentration of female hormones such as estrogen decreases, menopausal disorders are induced mainly in women in their late 40s to 50s. Among them, in women in their 30s to mid-40s, in addition to the symptoms of menopausal disorders described above, menstrual irregularities occur in addition to stress or excessive fatigue. This is called premenopausal disorders, and like menopausal disorders, it significantly reduces the QOL (quality of life) of women. In the treatment of these menopausal disorders or premenopausal disorders, hormone replacement therapy is mainly performed, but there are problems that it can cause cancer, or there are side effects such as irregular genital bleeding, vomiting, headache, liver dysfunction. For any woman, it is possible to induce premenopausal disorders or menopausal disorders, and in particular in an aging Japan, it is very important for women to transition from premenopausal to menopausal healthily. Therefore, like the treatment of infertility, a safe and effective treatment method for premenopausal disorders or menopausal disorders is being sought (Non-Patent Literature 2).

[0005] However, not only humans, but also for livestock such as cows or pigs, the aging of female reproductive tissues is gradually becoming a problem. For example, in the pig industry, the cycle from mating to delivery of a sow is 2.5 times per year, but as the age increases, the number of piglets born by the sow decreases, and the reproductive ability becomes poor due to an increase in reproductive disorders such as miscarriage. Therefore, it is desirable to develop a method that can suppress the aging process of female reproductive tissues in non-human animals in addition to humans.

[0006] Prior Art Documents

[0007] Non-patent literature

[0008] Non-patent literature 1: Jisan Fujin, 1998, Vol. 50, No. 6, N135-N138

[0009] Non-patent literature 2: JIM, 1999, Vol. 9, No. 8, 688-691 SUMMARY

[0010] (I) Technical problem to be solved

[0011] The present application has an object to provide an anti-aging agent for an ovary, follicle, or ovum (hereinafter sometimes collectively referred to as "female reproductive tissue").

[0012] (II) Technical solution

[0013] The present inventors have found that pyrroloquinoline quinone or a salt thereof can inhibit the aging process of a female reproductive tissue and can promote the production of estradiol. The present application has been completed on the basis of this new finding.

[0014] The present application provides, for example, the following inventions.

[0015] [1] An anti-aging agent for an ovary, follicle, or ovum, which contains pyrroloquinoline quinone or a salt thereof.

[0016] [2] An inhibitor or improver of fibrosis of a female reproductive tissue, which contains pyrroloquinoline quinone or a salt thereof.

[0017] [3] An ovum quality improver, which contains pyrroloquinoline quinone or a salt thereof.

[0018] [4] An improver or preventive of menstrual irregularity, which contains pyrroloquinoline quinone or a salt thereof.

[0019] [5] An improver or preventive against ovarian hyperstimulation syndrome, which contains pyrroloquinoline quinone or a salt thereof.

[0020] [6] An estrogen production promoter, which contains pyrroloquinoline quinone or a salt thereof.

[0021] [7] The estrogen production promoter according to [6], which ameliorates or prevents a symptom of a female due to a lack of estrogen.

[0022] [8] The estrogen production promoter according to [6], which ameliorates or prevents a premenopausal disorder or menopausal disorder of a female.

[0023] [9] The estrogen production promoter according to [6], which ameliorates or prevents a menopausal symptom of a female.

[0024]

[10] The preparation according to any one of [1] to [9], which is for a non-human animal.

[0025]

[11] A method of inhibiting the aging process of an ovary, follicle, or ovum, by taking a composition containing pyrroloquinoline quinone or a salt thereof.

[0026] [2-1] The anti-aging agent according to [1], which inhibits or improves fibrosis of female reproductive tissues.

[0027] [2-2] The anti-aging agent according to [1], which improves the quality of oocytes.

[0028] [2-3] The anti-aging agent according to [1], which ameliorates or prevents irregular menstruation.

[0029] [2-4] The anti-aging agent according to [1], which ameliorates or prevents ovarian hyperstimulation syndrome.

[0030] [2-5] The anti-aging agent according to [1], which promotes the production of estrogen.

[0031] [2-6] The anti-aging agent according to [2-5], which ameliorates or prevents symptoms caused by estrogen deficiency in females.

[0032] [2-7] The anti-aging agent according to [2-5], which ameliorates or prevents premenopausal or menopausal disorders in females.

[0033] [2-8] The anti-aging agent according to [2-5], which ameliorates or prevents menopausal symptoms in females.

[0034] (III) Beneficial Effects

[0035] The present application can provide an anti-aging agent for female reproductive tissues. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 is a graph showing the AMH concentration in the blood of each female mouse in Test Example 1.

[0037] Figure 2 (A) in is a graph showing the estrous cycle when no pyrroloquinoline quinone disodium salt was administered to the aged model mouse of a female in Test Example 2. Figure 2 (B) in is a graph showing the estrous cycle when pyrroloquinoline quinone was administered to the aged model mouse of a female in Test Example 2. In Figure 2 In, M indicates metestrus, WE indicates diestrus, E indicates estrus, P indicates proestrus, and D indicates dioestrus.

[0038] Figure 3(A) in the figure is a graph showing the relationship between the time of administration of eCG (equine chorionic gonadotropin) and the concentration of estradiol in blood in immature female mice given or not given disodium pyrroloquinoline quinone in Experiment 4. Figure 3 (B) in the figure shows the relationship between the time of administration of eCG (equine chorionic gonadotropin) and hCG (human chorionic gonadotropin) and the concentration of progesterone in the blood in immature female mice given or not given disodium pyrroloquinoline quinone in Experiment Example 4.

[0039] Figure 4 (A) is a photograph (20x magnification) taken of the ovaries of female-aged model mice before administration of disodium pyrroloquinoline quinone in Experiment Example 5 after PSR staining. Figure 4 (B) is a photograph (20x magnification) taken after PSR staining of the ovaries of female aging model mice given disodium pyrroloquinoline quinone for 2 weeks in Experiment Example 5. Figure 4 (C) and Figure 4 (D) in the text refers to... Figure 4 (A) and Figure 4 The diagram in (B) is tracing, and the areas marked with diagonal lines indicate the colored areas. Detailed Implementation

[0040] The specific embodiments of the present invention will be described in detail below. However, the present invention is not limited to the following embodiments.

[0041] [1. Anti-aging agents for the ovaries, follicles, or eggs]

[0042] The anti-aging agent for the ovaries, follicles, or eggs in this embodiment contains pyrroloquinoline quinone or a salt thereof (also known as “(A) component”).

[0043] Pyrroloquinoline quinone or a salt thereof exerts an effect of inhibiting or reducing disappearance of AMH (Anti-Mullerian Hormone) in females. Here, AMH is a hormone that is an index of aging of female reproductive tissues (particularly, ovary), and it is known that the concentration of AMH in blood decreases as the process of aging of female reproductive tissues progresses. In addition, pyrroloquinoline quinone or a salt thereof exerts an effect of inhibiting or improving fibrosis of female reproductive tissues (ovary, follicle, ovum, etc., particularly, ovary). Therefore, as one embodiment of the present application, there is provided an anti-aging agent for ovary, follicle, or ovum, and an inhibitor or improver of fibrosis of female reproductive tissues, which contains pyrroloquinoline quinone or a salt thereof. The "anti-aging" referred to here means that a woman regains youth, that the process of aging of a woman is inhibited, that a high QOL can be maintained even in an elderly woman, that an elderly woman can live with vitality and youth, and the like. In addition, the "elderly" referred to here means 30 years old or older, preferably 40 years old or older, and more preferably 50 years old or older. In addition, as one example of aging of female reproductive tissues (particularly, ovary tissues), the ovarian reserve function (the number of residual secondary follicles in the ovary) can be cited. As described above, since pyrroloquinoline quinone or a salt thereof has an effect of inhibiting or reducing disappearance of AMH in females, it also has an effect of improving the ovarian reserve function.

[0044] Pyrroloquinoline quinone or a salt thereof exerts an effect of improving the quality of ovum. Therefore, as one embodiment of the present application, there is provided an agent for improving the quality of ovum, which contains pyrroloquinoline quinone or a salt thereof. The "improvement of the quality of ovum" referred to here means that the number of ovulations is increased, and that the fertilization rate of the ovum discharged is maintained or improved. The "fertilization rate" referred to here means the proportion of the number of fertilized ovum to the total number of ovum discharged. Therefore, as one embodiment of the present application, there is provided an agent for increasing the number of ovum discharged and maintaining or improving the fertilization rate of ovum discharged, which contains pyrroloquinoline quinone or a salt thereof. In addition, the "improvement of the quality of ovum" means that the fertilization rate of ovum is maintained or improved, and that the development rate is maintained or improved. The "development rate" referred to here means the proportion of the number of fertilized ovum that become blastocyst stage embryos to the total number of fertilized ovum. Therefore, as one embodiment of the present application, there is provided an agent for maintaining or improving the fertilization rate of ovum and maintaining or improving the development rate, which contains pyrroloquinoline quinone or a salt thereof.

[0045] Pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing irregular menstruation. Therefore, as one embodiment of the present application, there is provided an improver or preventer of irregular menstruation, which contains pyrroloquinoline quinone or a salt thereof. The "irregular menstruation" referred to here means a state in which the menstrual cycle of females (if the female is a non-human animal in which menstruation is amenorrhea, then it means ovulation) is abnormal. For example, in the case of humans, the normal menstrual cycle is 25 to 38 days, and a menstrual cycle of 24 days or less or 39 days or more, or no menstruation for 3 months or more (amenorrhea) is regarded as irregular menstruation.

[0046] Pyrroloquinoline quinone or a salt thereof exerts an effect of promoting production of an estrogen (e.g., estrone, estradiol, estriol, and preferably estradiol). Therefore, as one embodiment of the present application, there is provided an estrogen production promoter containing pyrroloquinoline quinone or a salt thereof. Furthermore, pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing a symptom caused by estrogen deficiency in a female, an effect of improving or preventing a menopausal symptom in a female, an effect of improving or preventing a perimenopausal disorder in a female, or an effect of improving or preventing a menopausal disorder in a female, in addition to an effect of supplementing female hormones. Therefore, as one embodiment of the present application, there is provided an improver or preventer of a symptom caused by estrogen deficiency in a female, an improver or preventer of a menopausal symptom in a female, or an improver or preventer of a perimenopausal disorder or a menopausal disorder in a female, containing pyrroloquinoline quinone or a salt thereof.

[0047] Pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing ovarian hyperstimulation syndrome. Therefore, as one embodiment of the present application, there is provided an improver or preventer of ovarian hyperstimulation syndrome containing pyrroloquinoline quinone or a salt thereof. Here, "ovarian hyperstimulation syndrome" refers to a condition in which an ovary is swollen due to overstimulation by an ovulation-promoting agent, thereby causing various symptoms due to ascites or pleural effusion, and when the condition is severe, blood is concentrated, causing complications such as renal failure or thrombosis. As specific symptoms of ovarian hyperstimulation syndrome, there can be mentioned abdominal distension, nausea, rapid weight gain, decreased urine output, abdominal pain, diarrhea, and the like. Furthermore, pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing symptoms caused by ovarian hyperstimulation syndrome and complications of ovarian hyperstimulation syndrome by improving or preventing ovarian hyperstimulation syndrome. In addition, the preparations containing pyrroloquinoline quinone or a salt thereof of the above embodiments are collectively referred to as "preparations of the present embodiment".

[0048] In the present specification, "menopause" refers to a period of about 10 years before and after menopause in a female, and for humans, the period from the later 40s to the 50s is the menopausal period. Furthermore, in the present specification, "perimenopause" refers to a period younger than the above menopausal period, and is a period in which symptoms of a perimenopausal disorder described later appear, and for humans, the period from the 30s to the middle of the 40s is the perimenopausal period.

[0049] As symptoms of "female menopausal disorders", for example, dyspnea, dizziness on standing, paresthesia, stiffness of the limbs, edema, fatigue, tinnitus, hyperemia of the head, hot flashes, sweating, palpitation, excitement, insomnia, irritability, emotional instability, depression, easy weeping, low libido, anxiety, headache, shoulder stiffness, chest pain, low back pain, arthralgia, coldness of the extremities, itching, dryness of the skin or eyes, nausea, anorexia, abdominal pain, constipation, diarrhea, frequent urination, dyspareunia, vulvar discomfort, and the like can be listed.

[0050] As symptoms caused by a deficiency of estrogen in females, for example, menstrual abnormalities, symptoms of autonomic nervous disorders, symptoms of the nervous system, symptoms of atrophy of the urogenital organs, hyperlipidemia, arteriosclerosis, hypertension, cerebral apoplexy, osteoporosis, osteopenia, skin atrophy, arthralgia, dry mouth, changes in taste, sleep disorders, symptoms of the above-mentioned "female menopausal disorders", and the like can be listed.

[0051] As symptoms of "female premenopausal disorders (female young menopausal disorders)", in addition to the symptoms of the above-mentioned "female menopausal disorders", menstrual irregularities, amenorrhea, and the like can be listed.

[0052] "Female menopausal symptoms" refer to the symptoms of the above-mentioned "female menopausal disorders" and the symptoms of "female premenopausal disorders (female young menopausal disorders)". However, the symptoms can occur not only in "menopausal" or "premenopausal" women but also in young women (for example, women in their teens when menstruation begins). As "female menopausal symptoms", the symptoms can be relatively mild among the above-mentioned specific symptoms. For example, states in which hot flashes or sweating are felt by the subject but the frequency thereof is low, states in which depression or shoulder stiffness is relatively small, and the like can be listed.

[0053] Pyrroloquinoline quinone is a well-known compound represented by the following formula.

[0054] [Chemical Formula 1]

[0055]

[0056] As a salt of pyrroloquinoline quinone, for example, an alkali metal salt, an alkaline earth metal salt, and an ammonium salt can be listed. As an alkali metal salt, for example, a sodium salt, a potassium salt, and a lithium salt can be listed. As an alkaline earth metal salt, for example, a calcium salt and a magnesium salt can be listed.

[0057] As pyrroloquinoline quinone or a salt thereof, an alkali metal salt of pyrroloquinoline quinone is preferred, a sodium salt of pyrroloquinoline quinone is more preferred, and a disodium salt of pyrroloquinoline quinone is further preferred.

[0058] Pyrroloquinoline quinone or a salt thereof can be used as a commercially available product. In addition, as pyrroloquinoline quinone or a salt thereof, for example, crops containing pyrroloquinoline quinone or a salt thereof such as natto, soybean, cocoa powder, cocoa mass, cocoa, parsley, green pepper, and the like can be used.

[0059] For the content of the (A) component in the preparation of the present embodiment, the total content of the (A) component is preferably 0.01 to 30% by weight, more preferably 0.2 to 10% by weight, further preferably 1 to 5% by weight, and particularly preferably 1.3 to 3% by weight, based on the total amount of the preparation of the present embodiment, from the viewpoint of stability of the preparation to be administered, when used for humans. In addition, for the content of the (A) component in the preparation of the present embodiment, the total content of the (A) component is preferably 0.01 to 0.3% by weight, more preferably 0.02 to 0.2% by weight, based on the total amount of the preparation of the present embodiment, when used for non-human animals.

[0060] The daily intake amount of the (A) component for the preparation of the present embodiment can differ depending on the state (body weight, age, sex, and the like) of the individual who takes it, the form of the preparation, and the like, and for humans, the daily intake amount of the (A) component is preferably 7 to 100 mg, more preferably 10 to 40 mg, and further preferably 20 to 25 mg, from the viewpoint of ease of intake of a single amount, the effectiveness based on the physiological action of the (A) component, and safety.

[0061] The preparation of the present embodiment can further contain a polyamine (also referred to as a "(B) component") and a crude drug (also referred to as a "(C) component"). Thereby, the effects of the present application can be more remarkably exerted.

[0062] A polyamine is an aliphatic hydrocarbon containing two or more primary amino groups in a molecule. As specific examples of the polyamine, for example, putrescine, cadaverine, ethylenediamine, trimethylenediamine, hexamethylenediamine, spermidine, caldine, homospermidine, aminopropylcadaverine, spermine, thermine, thermospermine, canavalmine, aminopentyl norspermidine, aminopropyl homospermine, canavalmine, homospermine, caldopentamine, homocaldopentamine, aminopropyl canavalmine, bis(aminopropyl)homospermine, bis(aminopropyl)norspermine, aminobutyl canavalmine, aminopropyl homospermine, homopentamine, caldohexamine, homocaldohexamine, thermohexamine, homothermohexamine can be listed. From the viewpoint of easy use, as the polyamine, spermidine, spermine, putrescine are preferable, spermidine, spermine are more preferable, and spermidine is further preferable.

[0063] The polyamine can also be a commercially available product. The polyamine can be used singly or in combination of two or more. In addition, as the polyamine, an extract from a plant such as soybean (preferably soybean germ), wheat (preferably wheat germ), or rice (preferably rice germ), an extract from a fish or shellfish (preferably fish milt), or a polyamine-containing composition (also referred to as "polyamine composition") such as dried sake yeast can also be used. When the polyamine composition is an extract from a plant, water, or an organic solvent such as ethanol or a mixed solvent of these solvents, or the like can be used as an extraction solvent. From the viewpoint of ease of use, as the polyamine composition, an extract from a plant is preferred, and an extract from soybean (preferably soybean germ) is more preferred. As commercially available polyamine compositions, for example, "Soypolya" (manufactured by Combi Corporation), "ORYZAPOLYAMINE-P" (manufactured by Oryza Oil & Fat Chemical Co., Ltd.), "ORYZA POLYAMINE-LC" (manufactured by Oryza Oil & Fat Chemical Co., Ltd.), "PHYTOPOLYAMINE-S" (manufactured by TOYOBO CO., LTD.), "PHYTOPOLYAMINE-SP" (manufactured by TOYOBO CO., LTD.), "Elion SP" (manufactured by MITSUBISHI GAS CHEMICAL COMPANY, INC.), and the like can be exemplified.

[0064] The content of the (B) component in the preparation of the present embodiment is not particularly limited and can be appropriately set depending on the kind of the (B) component, the kinds and contents of other blended components, the form of the preparation, and the like. From the viewpoint of stability of the preparation, the viewpoint of reducing the influence of an odor possessed by the (B) component, and the like, the content of the (B) component is, for example, preferably 0.001 to 10% by mass, more preferably 0.01 to 0.1% by mass, and further preferably 0.02 to 0.05% by mass, based on the total amount of the preparation of the present embodiment.

[0065] In the preparation of the present embodiment, when a polyamine composition is contained as the (B) component, the content of the polyamine composition is not particularly limited and can be appropriately set depending on the kind of the polyamine composition, the kinds and contents of other blended components, the form of the preparation, and the like. From the viewpoint of stability of the preparation, the viewpoint of effectiveness based on the physiological action of the (B) component, the viewpoint of safety, the viewpoint of reducing the influence of an odor possessed by the (B) component, and the like, the content of the polyamine composition is, for example, preferably 5 to 35% by mass, more preferably 10 to 30% by mass, and further preferably 14 to 23% by mass, based on the total amount of the preparation of the present embodiment.

[0066] The content ratio of the (B) component with respect to the (A) component in the preparation of the present embodiment is not particularly limited and can be appropriately set in accordance with the kinds of the (A) component and the (B) component, the kinds and amounts of other blending components, the form of the preparation, and the like. From the viewpoint of further improving the effects brought about by the present application, for example, the total amount of the (B) component is preferably 0.001 to 0.1 parts by weight, more preferably 0.01 to 0.03 parts by weight, and further preferably 0.015 to 0.02 parts by weight, with respect to 1 part by weight of the total amount of the (A) component contained in the preparation of the present embodiment.

[0067] In the preparation of the present embodiment, when the polyamine composition is contained as the (B) component, the content ratio of the polyamine composition with respect to the (A) component is not particularly limited and can be appropriately set in accordance with the kinds of the (A) component and the polyamine composition, the kinds and amounts of other blending components, the form of the preparation, and the like. From the viewpoint of further improving the effects brought about by the present application, for example, the total amount of the polyamine composition is preferably 5 to 15 parts by weight, and more preferably 7 to 10 parts by weight, with respect to 1 part by weight of the total amount of the (A) component contained in the preparation of the present embodiment.

[0068] In the preparation of the present embodiment, when the (B) component contains spermidine and spermine, the weight ratio of spermine with respect to spermidine is not particularly limited and can be appropriately set in accordance with the kinds of the (A) component and the (B) component, the kinds and amounts of other blending components, the form of the preparation, and the like. From the viewpoint of further improving the effects brought about by the present application, for example, the weight ratio of spermine with respect to spermidine is preferably 0.01 to 10, more preferably 0.1 to 2, and further preferably 0.2 to 0.5.

[0069] In the preparation of the present embodiment, the daily intake amount of the (B) component varies depending on the state (body weight, age, sex, and the like) of the individual who takes it, the form of the preparation, and the like, but from the viewpoint of the effectiveness based on the physiological action of the (B) component, the viewpoint of safety, and the viewpoint of reducing the influence of the peculiar smell possessed by the (B) component, the daily intake amount of the (B) component is preferably 0.001 to 20 mg, more preferably 0.01 to 2 mg, further preferably 0.1 to 0.7 mg, and particularly preferably 0.3 to 0.4 mg.

[0070] In the preparation of the present embodiment, when the polyamine composition is contained as the (B) component, the daily intake amount of the polyamine composition varies depending on the state (body weight, age, sex, etc.) of the individual who takes it, the preparation form, etc., but from the viewpoint of ease of intake in a single amount, the effectiveness of the physiological action of the (B) component, the safety, and the viewpoint of reducing the influence of the odor possessed by the (B) component, it is preferably 50 to 400 mg, more preferably 100 to 300 mg, further preferably 200 to 250 mg, and particularly preferably 201 to 220 mg.

[0071] Crude drugs refer to all or a part of plants, animals, minerals, and the like natural products having medicinal purposes, or substances obtained by subjecting all or a part of the plants, animals, minerals, and the like natural products to simple processes such as drying. In the preparation of the present embodiment, as the form of crude drugs, for example, crude drugs themselves (raw drugs), crude drug powders obtained by drying and powdering raw drugs, crude drug extracts obtained by extracting raw drugs or crude drug powders with water, or organic solvents such as ethanol or mixed solvents of these solvents, and the like can be exemplified. Among them, from the viewpoint of more significantly exerting the effects brought about by the present application, crude drug extracts are preferred. The crude drug extracts can be infusions themselves (tinctures, fluid extracts, and the like), can be products obtained by diluting or concentrating infusions (viscous extracts, and the like), or can also be products obtained by drying infusions and powdering or pasting them (dry extracts, and the like).

[0072] Specific examples of herbal extracts include ginger extract, maca extract, licorice extract, astragalus extract, tangerine peel extract, peony extract, ginseng extract, turmeric extract, cistanche extract, cinnamon extract, eleutherococcus extract, hawthorn extract, rehmannia extract, angelica extract, saw palmetto extract, ginkgo leaf extract, bupleurum extract, cimicifuga extract, yam extract, jujube extract, atractylodes extract, lancea extract, ganoderma extract, deer antler extract, tongkat ali extract, French coastal pine bark extract, European oak extract, wild yam extract, yam extract containing diosgenin, cistanche tubulosa extract, garlic extract, eleutherococcus extract, verticillata extract, gardenia extract, shilajiri extract, cordyceps mycelium powder, black ginger extract, black pepper extract, pomegranate seed extract, vitex extract, black cimicifuga extract, and dandelion extract. From the perspective of formulation stability, as herbal extracts, maca extract, ginseng extract, saw palmetto extract, ginger extract, Tongkat Ali extract, French coastal pine bark extract, European oak extract, wild yam extract, yam extract containing diosgenin, pomegranate seed extract, and vitex extract are preferred, with maca extract and ginseng extract being even more preferred.

[0073] Commercially available products can also be used for raw medicinal herbs. A single raw medicinal herb can be used, or two or more can be used in combination.

[0074] The content of component (C) in the formulation of this embodiment is not particularly limited, and can be appropriately set according to the type of component (C), the type and content of other admixtures, the formulation form, etc. From the perspective of more significantly exerting the effects brought about by the present invention, the content of component (C), for example based on the total amount of the formulation of this embodiment, is preferably 0.1 to 70% by weight, more preferably 1 to 10% by weight, and even more preferably 2 to 4% by weight.

[0075] The content ratio of component (C) to component (A) in the formulation of this embodiment is not particularly limited, and can be appropriately set according to the types of components (A) and (C), the types and contents of other admixtures, the formulation form, etc. From the perspective of further improving the effect of the present invention, as the content ratio of component (C) to component (A), for example, relative to 1 part by weight of the total content of component (A) contained in the formulation of this embodiment, the total content of component (C) is preferably 0.1 to 3 parts by weight, more preferably 0.5 to 2.5 parts by weight, and even more preferably 1 to 1.5 parts by weight.

[0076] The content ratio of the (C) component with respect to the (B) component in the preparation of the present embodiment is not particularly limited and can be appropriately set according to the kinds of the (B) component and the (C) component, the kinds and contents of other blending components, the form of the preparation, and the like. From the viewpoint of further improving the effects brought by the present application, as the content ratio of the (C) component with respect to the (B) component, for example, with respect to 1 part by weight of the total content of the (B) component contained in the preparation of the present embodiment, the total content of the (C) component is preferably from 50 to 250 parts by weight, more preferably from 80 to 200 parts by weight, and further preferably from 100 to 170 parts by weight.

[0077] The preparation of the present embodiment can contain a pharmacologically active component or a physiologically active component other than the (A) component, the (B) component, and the (C) component, within a range that does not impair the effects of the present application. As specific examples of the pharmacologically active component or the physiologically active component, for example, ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, folic acid, vitamin B12, vitamin C, vitamin D, vitamin A, vitamin E, nicotinic acid, calcium pantothenate, taurine, carnosine, anserine, cetane, citrulline, gamma-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, carnitine, glutamic acid, glutamine, creatine, carnitine, luteolin, quercetin, genistein, cyanidin, resveratrol, diosgenin, isoflavone aglycone, isoflavone, lipoic acid, zinc, iron, calcium, selenium, astaxanthin, zeaxanthin, beta-carotene, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, placenta extract, pycnogenol (proanthocyanidin), equol, and the like can be listed. From the viewpoint of further improving the effects brought by the present application, as the pharmacologically active component or the physiologically active component, ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, resveratrol, isoflavone, equol, pycnogenol (proanthocyanidin), folic acid are preferred, and ubiquinone (coenzyme Q10), astaxanthin, zinc, resveratrol, isoflavone, equol, pycnogenol (proanthocyanidin), folic acid are more preferred. The pharmacologically active component or the physiologically active component can be used alone or in combination of two or more.

[0078] The preparation of the present embodiment can contain various additives other than the above-mentioned components within a range not impairing the effects of the present application. As specific examples of the additive, for example, excipients, binders, disintegrants, lubricants, coloring agents, flavoring agents, odor correcting agents, sugars, sugar alcohols / polyols, high-intensity sweeteners, oils, emulsifiers, thickening agents, acidifiers, fruit juices, and the like can be exemplified. As the excipient, lactose, white sugar, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, silicic acid, and the like can be exemplified. As the binder, water, ethanol, propanol, monosugar syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, calcium phosphate, polyvinylpyrrolidone, and the like can be exemplified. As the disintegrant, dried starch, sodium alginate, agar powder, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, glycerol monostearate, lactose, and the like can be exemplified. As the lubricant, talc, stearate, borax, polyethylene glycol, and the like can be exemplified. As the flavoring agent, white sugar, orange peel, citric acid, tartaric acid, and the like can be exemplified. As the sugar, sucrose, isomerized sugar, glucose, fructose, palatinose, trehalose, lactose, xylose, and the like can be exemplified. As the sugar alcohols / polyols, sorbitol, xylitol, erythritol, lactitol, isomalt, hydrogenated sugar syrup, hydrogenated malt sugar syrup, glycerol, propylene glycol, and the like can be exemplified. As the high-intensity sweetener, aspartame, stevia, acesulfame potassium, sucralose, and the like can be exemplified. As the oil, safflower oil, grape seed oil, sunflower oil, olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, perilla oil, and the like can be exemplified. As the emulsifier, sucrose fatty acid ester, glycerol fatty acid ester, lecithin, and the like can be exemplified. As the thickening agent, carrageenan, xanthan gum, guar gum, pectin, locust bean gum, and the like can be exemplified. As the acidifier, citric acid, lactic acid, malic acid, and the like can be exemplified. As the fruit juice, lemon juice, orange juice, berry juice, and the like can be exemplified. The additive can be used singly or in combination of two or more. From the viewpoint of the stability of the (A) component and the (B) component at the time of preparation, as the additive, it is preferable to contain an oil.

[0079] The dosage form of the preparation of the present embodiment is not particularly limited, and for example, oral preparations such as tablets (including oral disintegrating tablets, chewable tablets, lozenges, and the like), granules, powders, soft capsules, hard capsules, lozenges, jellies, or liquid preparations (including suspensions, emulsions, syrups, and the like), and the like; external preparations such as ointments, suppositories, patches, sprays, and the like; injections, and the like can be exemplified. Among them, from the viewpoint of easy handling, further improvement of the effects brought by the present application, oral preparations are preferable, and tablets, granules, soft capsules, hard capsules are more preferable.

[0080] The preparation of the present embodiment can be used, for example, as an ingredient of a medicine, a quasi-drug, a cosmetic, a food (beverage, food). In addition, the preparation of the present embodiment can be used, for example, as a pharmaceutical preparation, a quasi-drug preparation, a specific health food, a nutritionally functional food, a food for the elderly, a food for a specific use, a functional food, a health supplement (nutritional supplement), a food preparation (e.g., a candy tablet), a food for obvious use. Further, the preparation of the present embodiment can also be used, for example, as an animal medicine, a feed additive.

[0081] When the preparation of the present embodiment is used as a food, the food can be a food in which the (A) component and, as necessary, other components are mixed in a general food. As such a food, solid foods such as cookies, biscuits, snack bars, jellies, gummies, chocolates, chewing gums, caramels, cheeses, and the like; and liquid foods such as nutritional beverages, fruit juices, tea beverages, coffee beverages, milk beverages, and the like can be exemplified.

[0082] The preparation of the present embodiment can be suitably used for a subject (e.g., a human; a non-human animal such as a cow, a pig, a horse, a sheep, a goat, a dog, a rabbit, a mouse, a rat, a guinea pig, a gerbil, a hamster, a marmoset, and the like) in which an action of inhibiting the aging process of female reproductive tissues is required. From the viewpoint of further improving the effects brought by the present application, as the subject, a human is preferred. In addition, from the viewpoint of further improving the effects brought by the present application, among non-human animals, a cow, a pig, or a horse is preferred, and a pig is more preferred.

[0083] [2. A method of inhibiting the aging process of an ovary, a follicle, or an ovum]

[0084] Pyrroloquinoline quinone or a salt thereof has an action of inhibiting the aging process of an ovary, a follicle, or an ovum. Therefore, as one embodiment of the present application, a method of inhibiting the aging process of an ovary, a follicle, or an ovum by taking a composition containing pyrroloquinoline quinone or a salt thereof is provided. The ovary, the follicle, or the ovum can be a human ovary, a human follicle, or a human ovum, or a non-human animal ovary, a non-human animal follicle, or a non-human animal ovum such as a cow ovary, a cow follicle, or a cow ovum, a pig ovary, a pig follicle, or a pig ovum, a horse ovary, a horse follicle, or a horse ovum, a sheep ovary, a sheep follicle, or a sheep ovum, a goat ovary, a goat follicle, or a goat ovum, a dog ovary, a dog follicle, or a dog ovum, a rabbit ovary, a rabbit follicle, or a rabbit ovum, a mouse ovary, a mouse follicle, or a mouse ovum, a rat ovary, a rat follicle, or a rat ovum, a guinea pig ovary, a guinea pig follicle, or a guinea pig ovum, a gerbil ovary, a gerbil follicle, or a gerbil ovum, a hamster ovary, a hamster follicle, or a hamster ovum, a marmoset ovary, a marmoset follicle, or a marmoset ovum, and the like. From the viewpoint of further improving the effects brought by the present application, a human ovary, a human follicle, or a human ovum is preferred. In addition, from the viewpoint of further improving the effects brought by the present application, among non-human animal ovaries, follicles, and ova, a cow ovary, a cow follicle, or a cow ovum, a pig ovary, a pig follicle, or a pig ovum, or a horse ovary, a horse follicle, or a horse ovum is preferred, and a pig ovary, a pig follicle, or a pig ovum is more preferred.

[0085] In addition, as to the kind, content, and daily intake amount of pyrroloquinoline quinone or a salt thereof in the composition in these embodiments, the kind, content, and daily intake amount of other components, and the like, and the preparation form of the composition, and the like, the explanations in [1. Anti-aging agent for an ovary, a follicle, or an ovum] apply.

[0086] Examples

[0087] Hereinafter, the present application is more specifically described based on examples and the like. However, the present application is not limited by the following examples. In addition, the following test examples are all conducted with the approval of the Animal Experiment Ethics Committee of Hiroshima University.

[0088] [Test Example 1: Effect of administration of pyrroloquinoline quinone on AMH (anti-Mullerian hormone) in blood of female mice]

[0089] Six-month-old Leydig cell-specific NRG1 -deleted female mice were administered 2 weeks of PQQ disodium salt (2 mg / kg / day) through drinking water (n = 5). In addition, six-month-old Leydig cell-specific NRG1 -deleted male mice (KO; control group) and six-month-old wild-type mice (WT mice) were each administered 2 weeks of water only. Thereafter, blood was recovered, and the concentration of AMH (anti-Mullerian hormone), which is a marker of ovarian follicles, was measured using an AMH ELISA kit (Elabscience Biotechnology, Bethesda, MD) and in accordance with the method for use of the kit. The results are shown in Figure 1 In addition, the lifespan of Leydig cell-specific NRG1 -deleted female mice is short, and six months of age corresponds to more than 40 years of age in humans.

[0090] As shown in Figure 1 , the concentration of AMH decreased in the control group of female mice not administered PQQ disodium salt compared to WT mice, whereas the concentration of AMH increased in female mice administered PQQ disodium salt compared to WT mice. That is, it was confirmed that by ingesting PQQ disodium salt, the ovarian tissue was rejuvenated.

[0091] [Test Example 2: Effect of administration of pyrroloquinoline quinone on estrous cycle of female mice]

[0092] Six-month-old female Reye's cell-specific NRG1-deficient mice (KO) were continuously administered PQQ disodium salt (2 mg / kg / day) via drinking water. Female mice given only water served as a control group. Estrogenic cycles were assessed using vaginal smears. Vaginal smears were examined as follows: physiological saline was refluxed into the vagina, and the refluxate was smeared onto a glass slide. After drying, the slide was fixed with methanol and stained with Giemsa stain. The slide was observed under an inverted microscope (manufactured by KEYENCE CORPORATION). Anestrus was defined as the presence of only white blood cells, proestrus as the presence of only nucleated epithelial cells, estrus as the presence of only anucleated epithelial cells, microestrus as the presence of a mixture of anucleated epithelial cells and white blood cells with a higher number of anucleated epithelial cells, and metestrus as the presence of a mixture of anucleated epithelial cells and white blood cells with a higher number of white blood cells. The results are presented below. Figure 2 .

[0093] like Figure 2 As shown, it can be confirmed that the female mice in the control group, which were not given PQQ disodium salt, remained in a microestrus state throughout, and no cyclical changes in estrus were observed. In contrast, the female mice given PQQ disodium salt showed cyclical changes in estrus again approximately 14 days after the start of administration. That is, it can be confirmed that the abnormal ovulation cycle in female mice was improved by administering PQQ disodium salt.

[0094] [Experiment 3: Effect of pyrroloquinoline quinone administration on oocytes]

[0095] Three-week-old immature female mice were administered PQQ disodium salt (2 mg / kg / day) via drinking water (n=5). A control group of female mice was given only water. Two days after the start of administration, 4 IU of eCG (equine chorionic gonadotropin) to induce follicle development was administered, followed by 5 IU of hCG (human chorionic gonadotropin) 48 hours later. The number of ovulations was then determined for each female mouse. The number of ovulations was calculated by retrieving the oviduct from the slaughtered mouse, cutting open the ampulla of the oviduct, and recovering the released oocytes and cumulus cell complexes. The complexes were observed visually under a stereomicroscope. Furthermore, the released oocytes were used for in vitro fertilization, and the fertilization rate and development rate were calculated. Here, a fertilized oocyte is a divided oocyte 24 hours after fertilization, and the fertilization rate is the ratio of the number of successfully fertilized oocytes to the total number of mature oocytes released. Furthermore, the development rate refers to the proportion of fertilized eggs that develop into blastocysts. The number of ovulations and fertilization rate are presented in Table 1, expressed as mean ± standard deviation. Additionally, for mice, 3 weeks of age corresponds to the teenage years at which menstruation begins in humans.

[0096] [Table 1]

[0097]

[0098] As shown in Table 1, the number of ovulations of the female mice administered with PQQ disodium salt increased while maintaining the in vitro fertilization rate, and the development rate increased while maintaining the in vitro fertilization rate. That is, it was confirmed that the quality of the ovum increased by administering PQQ disodium salt.

[0099] [Experiment 4: Effect of administration of pyrroloquinoline quinone on the concentration of female hormone in blood]

[0100] PQQ disodium salt (2 mg / kg / day) was administered to immature female mice of 3 weeks old through drinking water. In addition, female mice administered with water alone were used as a control group. At the time point after 2 days from the start of administration, eCG (equine chorionic gonadotropin) 4 IU was intraperitoneally administered to induce follicular development, followed by intraperitoneal administration of hCG (human chorionic gonadotropin) 5 IU after 48 hours. Serum of each female mouse was recovered over time from just before the administration of eCG, and the concentration of estradiol (E2) in blood, which is a marker of follicular development, and the concentration of progesterone (P4) in blood, which is a marker of corpus luteum, were measured using a commercially available kit of Endocrine Technology, Inc. (Newark, CA), respectively. The results are shown in Figure 3 .

[0101] As shown in (A) of Figure 3 , the concentration of estradiol in blood of the female mice administered with PQQ disodium salt increased over time compared to the control group of female mice not administered with PQQ disodium salt. That is, it was confirmed that the production of estradiol was promoted by administering PQQ disodium salt. It is well known that estradiol is one of female hormones widely used in hormone replacement therapy, which is used for the treatment of perimenopausal disorder or the improvement of menopausal symptoms. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof can be used as an improver or preventer of symptoms caused by a deficiency of estrogen, an improver or preventer of perimenopausal disorder or menopausal disorder of females. Thereby, a woman in perimenopause or menopause can come to amenorrhea without pain.

[0102] In addition, as shown in (B) of Figure 3 , no large difference was found in the increase in the concentration of progesterone in blood between the female mice administered with PQQ disodium salt and the control group of female mice not administered with PQQ disodium salt. It is known that progesterone is associated with the concentration of blood in ovarian hyperstimulation syndrome. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof can be used as a preventer of ovarian hyperstimulation syndrome by promoting the production of estradiol and alleviating the production of progesterone.

[0103] [Experiment Example 5: Effect of administration of pyrroloquinoline quinone on ovarian fibrosis]

[0104] A 6-month-old female mouse lacking NRG1 specific to Leydig cells (KO) was administered 2 weeks of PQQ disodium salt (2 mg / kg / day) through drinking water. Then, the excised ovary was subjected to PSR staining, and observed at 20 times magnification using an inverted microscope (manufactured by KEYENCE CORPORATION). The results are shown in Figure 4 .

[0105] As shown in Figure 4 , the collagen deposition, i.e., fibrosis of the ovary, was confirmed by the red staining of the stromal cells of the mouse before administration of PQQ disodium salt. In addition, atrophy of the stromal cells was also observed. On the other hand, for the mouse administered 2 weeks of PQQ disodium salt, disappearance of the red color of the stromal cells was confirmed, and the fibrotic stromal cells were improved to a healthy state. It is known that the stromal cells of the ovary gradually fibrose with aging. That is, it was confirmed that by taking PQQ disodium salt, the ovarian tissue was rejuvenated. In addition, cystic follicles were observed, and it was confirmed that atrophy of the follicles was also improved, and thus it was confirmed that the quality of the oocytes was improved.

[0106] According to the results of Experiment Examples 1 to 5, by taking pyrroloquinoline quinone or a salt thereof, it is possible to ensure the QOL of women who are easily damaged due to physical and mental abnormalities caused by perimenopause, and it is possible to make the elderly women also live with vitality and youth.

Claims

1. Use of pyrroloquinoline quinone or a salt thereof in the manufacture of an inhibitor or improver of fibrosis of female reproductive tissue.

2. Use of pyrroloquinoline quinone or a salt thereof in the manufacture of an improver or preventer of menstrual irregularity.

3. Use of pyrroloquinoline quinone or a salt thereof in the manufacture of an improver or preventer against ovarian hyperstimulation syndrome.

4. Use of pyrroloquinoline quinone or a salt thereof in the manufacture of an improver of abnormal ovulation cycle.

5. The use according to any one of claims 1 to 4, which improves or prevents a perimenopausal disorder or menopausal disorder in a female.

6. The use according to any one of claims 1 to 4, which improves or prevents menopausal symptoms in a female.

7. The use according to any one of claims 1 to 4, which is used for improving the fertility of a female.

Images