Female Anti-Aging Agent

Abstract

The present invention pertains to an anti-aging agent for ovaries, ovarian follicles or ova, said anti-aging agent comprising pyrroloquinoline quinone or a salt thereof.

Description

[Technical Field]

[0001] The present invention relates to female anti-aging agents. [Background technology]

[0002] As women age, reproductive tissues such as the ovaries, follicles, and eggs undergo aging, making it difficult to conceive and give birth. In infertility treatment for women who have difficulty conceiving, hormone replacement therapy is often used to induce ovulation. However, this method not only makes it difficult to achieve uniformly mature follicle development and ovulation, but also causes a side effect known as ovarian hyperstimulation syndrome (OHSS), which, if aggravated, can lead to respiratory failure and circulatory failure. Therefore, safe and effective infertility treatments are needed (Non-Patent Document 1).

[0003] Furthermore, as reproductive tissues in women age, the concentration of estrogen and other female hormones declines, primarily causing menopausal symptoms in women in their late 40s to 50s. Even women in their 30s to mid-40s may experience menstrual irregularities in addition to the aforementioned menopausal symptoms, due to stress and overwork. This condition, also known as premenopausal symptoms, significantly reduces women's quality of life (QOL) along with menopausal symptoms. Hormone replacement therapy (HRT) is commonly used to treat these menopausal and premenopausal disorders, but it carries the risk of side effects such as carcinogenesis, irregular vaginal bleeding, vomiting, headache, and liver dysfunction. While premenopausal and premenopausal disorders can occur in any woman, maintaining a healthy lifestyle from premenopause through menopause is crucial, especially in Japan, where the population is rapidly aging. Therefore, safe and effective treatments for premenopausal and premenopausal disorders, as well as infertility treatments, are needed (Non-Patent Document 2).

[0004] Aging of female reproductive tissues is a problem not only in humans but also in livestock such as cattle and pigs. For example, in the pig farming industry, the cycle from mating to parturition for sows is 2.5 times per year. However, as sows age, the number of offspring they give birth to decreases, and reproductive performance deteriorates due to an increase in reproductive disorders such as miscarriage. Therefore, there is a need to develop methods to suppress the progression of aging of female reproductive tissues in both humans and non-human animals. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Journal of the Japan Society of Obstetrics and Gynecology, 1998, Vol. 50, No. 6, N135-N138 [Non-patent document 2] JIM, 1999, Vol. 9, No. 8, 688-691 Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide an anti-aging agent for ovaries, follicles, or eggs (hereinafter, these may be collectively referred to as "female reproductive tissues"). [Means for solving the problem]

[0007] The present inventors have found that pyrroloquinoline quinone or a salt thereof inhibits the aging process in female reproductive tissues and promotes estradiol production, etc. The present invention is based on this novel finding.

[0008] The present invention provides, for example, the following inventions. [1] An anti-aging agent for ovaries, follicles or eggs, which contains pyrroloquinoline quinone or a salt thereof. [2] An agent for suppressing or improving fibrosis of female reproductive tissues, comprising pyrroloquinoline quinone or a salt thereof. [3] An agent for improving egg quality, containing pyrroloquinoline quinone or a salt thereof. [4] An agent for improving or preventing menstrual irregularities, comprising pyrroloquinoline quinone or a salt thereof. [5] An agent for improving or preventing ovarian hyperstimulation syndrome, comprising pyrroloquinoline quinone or a salt thereof. [6] An estrogen production promoter containing pyrroloquinoline quinone or a salt thereof. [7] The agent described in [6], which improves or prevents symptoms resulting from estrogen deficiency in females. [8] The agent described in [6], which improves or prevents premenopausal symptoms or menopausal symptoms in females. [9] The agent described in [6], which improves or prevents menopausal symptoms in females.

[10] The agent according to any one of [1] to [9], for use in non-human animals.

[11] A method for inhibiting the progression of aging of ovaries, follicles, or eggs by ingesting a composition containing pyrroloquinoline quinone or a salt thereof. [2-1] The agent described in [1], which suppresses or improves fibrosis of female reproductive tissues. [2-2] The agent described in [1] improves the quality of eggs. [2-3] The agent described in [1], which improves or prevents menstrual irregularities. [2-4] The agent described in [1], which improves or prevents ovarian hyperstimulation syndrome. [2-5] The agent described in [1], which promotes the production of estrogen. [2-6] The agent described in [2-5], which improves or prevents symptoms resulting from estrogen deficiency in females. [2-7] The agent described in [2-5], which improves or prevents premenopausal symptoms or menopausal symptoms in females. [2-8] The agent described in [2-5], which improves or prevents menopausal symptoms in females. [Effects of the Invention]

[0009] According to the present invention, an anti-aging agent for female reproductive tissues can be provided. [Brief explanation of the drawings]

[0010] [Figure 1] 1 is a graph showing the blood AMH concentration in each female mouse in Test Example 1. [Figure 2] 2A is a graph showing the estrous cycle when pyrroloquinoline quinone disodium salt is not administered to female aging model mice in Test Example 2. 2B is a graph showing the estrous cycle when pyrroloquinoline quinone is administered to female aging model mice in Test Example 2. In FIG. 2, M indicates metestrus, WE indicates microestrus, E indicates estrus, P indicates proestrus, and D indicates diestrus. [Figure 3] Graph (A) shows the relationship between time and blood estradiol concentration when eCG (pregnant mare serum gonadotropin) was administered to immature female mice administered with or without pyrroloquinoline quinone disodium salt in Test Example 4. Graph (B) shows the relationship between time and blood progesterone concentration when eCG (pregnant mare serum gonadotropin) and hCG (human chorionic gonadotropin) were administered to immature female mice administered with or without pyrroloquinoline quinone disodium salt in Test Example 4. [Figure 4] (A) is a photograph (magnification 20x) of PSR-stained ovaries of a female aging model mouse before administration of pyrroloquinoline quinone disodium salt in Test Example 5. (B) is a photograph (magnification 20x) of PSR-stained ovaries of a female aging model mouse after administration of pyrroloquinoline quinone disodium salt for two weeks in Test Example 5. (C) and (D) are traces of (A) and (B), respectively, and the shaded areas indicate the stained areas. DETAILED DESCRIPTION OF THE INVENTION

[0011] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The following describes in detail the embodiments of the present invention, but the present invention is not limited to the following embodiments.

[0012] [1. Anti-aging agents for ovaries, follicles, or eggs] The anti-aging agent for ovaries, follicles, or ova according to this embodiment contains pyrroloquinoline quinone or a salt thereof (also referred to as "component (A)").

[0013] Pyrroloquinoline quinone or a salt thereof has the effect of suppressing the disappearance or decrease of AMH (Anti-Mullerian Hormone) in females. AMH is a hormone that serves as an indicator of aging in female reproductive tissues (particularly the ovaries), and it is known that blood AMH concentrations decrease as aging of female reproductive tissues progresses. Furthermore, pyrroloquinoline quinone or a salt thereof has the effect of suppressing or ameliorating fibrosis in female reproductive tissues (ovaries, follicles, ova, etc., particularly the ovaries). Thus, one embodiment of the present invention provides an anti-aging agent for ovaries, follicles, or ova, and an agent for suppressing or ameliorating fibrosis in female reproductive tissues, each containing pyrroloquinoline quinone or a salt thereof. Here, "anti-aging" refers to rejuvenating women, suppressing the progression of female aging, enabling women to maintain a high quality of life even at an advanced age, and enabling women to live vibrantly and youthfully even at an advanced age. Furthermore, the term "elderly" as used herein refers to those in their 30s or older, preferably those in their 40s or older, and more preferably those in their 50s or older. One example of aging of female reproductive tissues (particularly ovarian tissues) is ovarian reserve (the number of secondary follicles remaining in the ovaries). As described above, pyrroloquinoline quinone or a salt thereof has the effect of suppressing the disappearance or decrease of AMH in females, and therefore also has the effect of improving ovarian reserve.

[0014] Pyrroloquinoline quinone or a salt thereof has the effect of improving egg quality. Therefore, one embodiment of the present invention provides an agent for improving egg quality, comprising pyrroloquinoline quinone or a salt thereof. Here, "improving egg quality" refers to increasing the number of ovulations while maintaining or improving the fertilization rate of the ovulated eggs, where the fertilization rate is the ratio of the number of fertilized eggs to the total number of ovulated eggs. Therefore, one embodiment of the present invention provides an agent for increasing the number of ovulated eggs and maintaining or improving the fertilization rate of the ovulated eggs, comprising pyrroloquinoline quinone or a salt thereof. Furthermore, "improving egg quality" refers to maintaining or improving the development rate while maintaining or improving the fertilization rate of eggs, where the development rate refers to the proportion of fertilized eggs that develop into blastocyst-stage embryos. Therefore, one embodiment of the present invention provides an agent for maintaining or improving the fertilization rate of eggs and maintaining or improving the development rate, comprising pyrroloquinoline quinone or a salt thereof.

[0015] Pyrroloquinoline quinone or a salt thereof has the effect of improving or preventing menstrual irregularities. Thus, one embodiment of the present invention provides an agent for improving or preventing menstrual irregularities, which contains pyrroloquinoline quinone or a salt thereof. Here, "menstrual irregularities" refers to a state in which the menstrual cycle in females (ovulation in non-menstruating female non-human animals) is abnormal. For example, in humans, a normal menstrual cycle is considered to be 25 to 38 days, and menstrual irregularities are defined as a menstrual cycle of 24 days or less or 39 days or more, or an absence of menstruation for 3 months or more (amenorrhea).

[0016] Pyrroloquinoline quinone or a salt thereof has the effect of promoting the production of estrogen (including estrone, estradiol, and estriol, preferably estradiol). Thus, one embodiment of the present invention provides an estrogen production promoter containing pyrroloquinoline quinone or a salt thereof. Furthermore, pyrroloquinoline quinone or a salt thereof promotes the production of estrogen, thereby improving or preventing symptoms resulting from estrogen deficiency in females, improving or preventing female menopausal symptoms, improving or preventing female premenopausal disorders, or improving or preventing female menopausal disorders, as well as the effect of supplementing female hormones. Thus, one embodiment of the present invention provides an agent for improving or preventing symptoms resulting from estrogen deficiency in females, an agent for improving or preventing female menopausal symptoms, or an agent for improving or preventing female premenopausal disorders or menopausal disorders, which contains pyrroloquinoline quinone or a salt thereof.

[0017] Pyrroloquinoline quinone or a salt thereof has the effect of improving or preventing ovarian hyperstimulation syndrome (OVHS). Accordingly, one embodiment of the present invention provides an agent for improving or preventing OVHS, containing pyrroloquinoline quinone or a salt thereof. Here, "ovarian hyperstimulation syndrome" refers to various symptoms caused by ascites and pleural effusion due to ovarian swelling caused by excessive stimulation by ovulation-inducing agents. In severe cases, blood may become concentrated, leading to complications such as renal failure and thrombosis. Specific symptoms of OVHS include abdominal distension, nausea, rapid weight gain, decreased urine output, abdominal pain, diarrhea, etc. Furthermore, pyrroloquinoline quinone or a salt thereof improves or prevents OVHS, thereby improving or preventing symptoms and complications associated with OVHS. The agents containing pyrroloquinoline quinone or a salt thereof according to the above-described embodiments are collectively referred to as "agents according to the present embodiments."

[0018] As used herein, "menopause" refers to the approximately 10 years before and after menopause in females, and in humans, the period from the late 40s to the 50s. Furthermore, as used herein, "pre-menopause" refers to a period younger than the menopause described above, during which symptoms of pre-menopausal disorders, described below, may occur, and in humans, the period from the 30s to the mid-40s corresponds to the pre-menopausal period.

[0019] Symptoms of "female menopause" include, for example, shortness of breath, lightheadedness, dizziness, numbness in the limbs, stiffness in the limbs, swelling, fatigue, tinnitus, hot flashes, sweating, palpitations, excitement, insomnia, irritability, anger, emotional instability, depression, tearfulness, loss of motivation, anxiety, headache, stiff shoulders, chest pain, back pain, joint pain, cold hands and feet, itching, dry skin and eyes, nausea, loss of appetite, abdominal pain, constipation, diarrhea, frequent urination, sexual dysfunction, and discomfort in the vulva.

[0020] Examples of "symptoms resulting from estrogen deficiency in females" include menstrual irregularities, autonomic nervous system symptoms, neuropsychiatric symptoms, atrophy of the urogenital organs, hyperlipidemia, arteriosclerosis, hypertension, stroke, osteoporosis, osteopenia, skin atrophy, joint pain, dry mouth, changes in taste, sleep disorders, and the symptoms of "female menopausal disorders" mentioned above, as well as the symptoms described in "1. Expected actions and effects of HRT" in the "Guidelines for Hormone Replacement Therapy 2017 Edition" edited by the Japan Society of Obstetrics and Gynecology.

[0021] Symptoms of "female pre-menopause (female premature menopause)" include irregular menstruation and amenorrhea in addition to the symptoms of "female menopause" mentioned above.

[0022] "Female menopausal symptoms" refers to the symptoms of "female menopausal disorder" and "female pre-menopausal disorder (premature female menopausal disorder)" described above. However, these symptoms are not limited to "menopause" or "pre-menopause" and can appear at an even younger age (for example, in humans, in the teens when menstruation begins). "Female menopausal symptoms" may also refer to milder symptoms of these specific symptoms described above. Examples include a state in which hot flashes and sweating are noticed but occur infrequently, and mild depression and stiff shoulders.

[0023] Pyrroloquinoline quinone has the following formula: [ka] It is a known compound represented by the formula:

[0024] Examples of salts of pyrroloquinoline quinone include alkali metal salts, alkaline earth metal salts, and ammonium salts. Examples of alkali metal salts include sodium salts, potassium salts, and lithium salts. Examples of alkaline earth metal salts include calcium salts and magnesium salts.

[0025] 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 even more preferred.

[0026] Commercially available pyrroloquinoline quinone or a salt thereof may be used. Alternatively, pyrroloquinoline quinone or a salt thereof may be used from, for example, natto, soybeans, cocoa powder, cacao mass, cacao, parsley, bell peppers, or other products containing pyrroloquinoline quinone or a salt thereof.

[0027] When used in humans, the content of component (A) in the agent according to this embodiment is preferably 0.01 to 30 wt%, more preferably 0.2 to 10 wt%, even more preferably 1 to 5 wt%, and particularly preferably 1.3 to 3 wt%, based on the total amount of the agent according to this embodiment, from the viewpoint of the stability of the administered formulation. When used in non-human animals, the content of component (A) in the agent according to this embodiment is preferably 0.01 to 0.3 wt%, more preferably 0.02 to 0.2 wt%, based on the total amount of the agent according to this embodiment.

[0028] In the agent according to this embodiment, the daily intake of component (A) may vary depending on the condition of the individual taking it (body weight, age, sex, etc.), the formulation, etc., but when used in humans, it is preferably 7 to 100 mg, more preferably 10 to 40 mg, and even more preferably 20 to 25 mg, from the viewpoints of ease of ingestion as a single dose, the effectiveness of the physiological action based on component (A), and safety, and when used in non-human animals, it is preferably 1 to 30 mg / kg, more preferably 2 to 20 mg / kg.

[0029] The agent according to this embodiment may further contain a polyamine (also referred to as "component (B)") and a herbal medicine (also referred to as "component (C)"). This makes the effects of the present invention more pronounced.

[0030] Polyamines are aliphatic hydrocarbons containing two or more primary amino groups in the molecule. Specific examples of polyamines include putrescine, cadaverine, ethylenediamine, trimethylenediamine, hexamethylenediamine, spermidine, caldine, homospermidine, aminopropylcadaverine, spermine, thermine, thermospermine, canavalmine, aminopentylnorspermidine, aminopropylhomospermine, canavalmine, homospermine, caldopentamine, homocaldopentamine, aminopropylcanavalmine, bis(aminopropyl)homospermine, bis(aminopropyl)norspermine, aminobutylcanavalmine, aminopropylhomospermine, homopentamine, caldohexamine, homocaldohexamine, thermohexamine, and homothermohexamine. From the viewpoint of ease of use, spermidine, spermine, and putrescine are preferred as polyamines, spermidine and spermine are more preferred, and spermidine is even more preferred.

[0031] Commercially available polyamines can also be used. One type of polyamine may be used alone, or two or more types may be used in combination. Furthermore, compositions containing polyamines (also referred to as "polyamine compositions") such as extracts from plants such as soybeans (preferably soybean germ), wheat (preferably wheat germ), or rice (preferably rice germ), extracts from seafood (preferably milt), or dried sake yeast can also be used as polyamines. When the polyamine composition is an extract from a plant, the extraction solvent can be water, an organic solvent such as ethanol, or a mixture thereof. From the viewpoint of ease of use, the polyamine composition is preferably an extract from a plant, and more preferably an extract from soybeans (preferably soybean germ). Examples of commercially available polyamine compositions include "SOYPOLIA" (manufactured by Combi Corporation), "ORYZA POLYAMINE-P" (manufactured by Oryza Oil & Fat Chemical Co., Ltd.), "ORYZA POLYAMINE-LC" (manufactured by Oryza Oil & Fat Chemical Co., Ltd.), "PHYTO POLYAMINE-S" (manufactured by Toyobo Co., Ltd.), "PHYTO POLYAMINE-SP" (manufactured by Toyobo Co., Ltd.), and "ELION SP" (manufactured by Mitsubishi Gas Chemical Co., Inc.).

[0032] The content of component (B) in the agent according to this embodiment is not particularly limited and is set appropriately depending on the type of component (B), the types and contents of other blended components, the formulation form, etc. From the viewpoint of formulation stability and minimizing the effect of the odor of component (B), the content of component (B) is, for example, preferably 0.001 to 10% by weight, more preferably 0.01 to 0.1% by weight, and even more preferably 0.02 to 0.05% by weight, based on the total amount of the agent according to this embodiment.

[0033] When the agent according to this embodiment contains a polyamine composition as component (B), the content of the polyamine composition is not particularly limited and is set appropriately depending on the type of polyamine composition, the types and contents of other blended components, the formulation form, etc. From the viewpoints of formulation stability, the effectiveness of the physiological action based on component (B), safety, and reducing the influence of the odor of component (B), the content of the polyamine composition is, for example, preferably 5 to 35 wt %, more preferably 10 to 30 wt %, and even more preferably 14 to 23 wt %, based on the total amount of the agent according to this embodiment.

[0034] The content ratio of component (B) to component (A) in the agent according to this embodiment is not particularly limited and is set appropriately depending on the types of components (A) and (B), the types and contents of other blended components, the dosage form, etc. From the viewpoint of further enhancing the effects of the present invention, the content ratio of component (B) to component (A) is, for example, preferably 0.001 to 0.1 parts by weight, more preferably 0.01 to 0.03 parts by weight, and even more preferably 0.015 to 0.02 parts by weight, of the total content of component (B) per 1 part by weight of the total content of component (A) contained in the agent according to this embodiment.

[0035] When the agent according to the present embodiment contains a polyamine composition as component (B), the content ratio of the polyamine composition relative to component (A) is not particularly limited and is set appropriately depending on the types of component (A) and polyamine composition, the types and contents of other blended components, the formulation form, etc. From the viewpoint of further enhancing the effects of the present invention, the content ratio of the polyamine composition relative to component (A) is, for example, preferably 5 to 15 parts by weight, and more preferably 7 to 10 parts by weight, of the total content of the polyamine composition relative to 1 part by weight of the total content of component (A) contained in the agent according to the present embodiment.

[0036] In the agent according to this embodiment, when component (B) contains spermidine and spermine, the weight ratio of spermine to spermidine is not particularly limited and is appropriately set depending on the types of components (A) and (B), the types and contents of other blended components, the formulation form, etc. From the viewpoint of further enhancing the effects of the present invention, the weight ratio of spermine to spermidine is, for example, preferably 0.01 to 10, more preferably 0.1 to 2, and even more preferably 0.2 to 0.5.

[0037] In the agent according to this embodiment, the daily intake of component (B) may vary depending on the condition of the individual taking it (body weight, age, sex, etc.), the formulation, etc., but from the viewpoints of the effectiveness of the physiological action of component (B), safety, and minimizing the impact of the odor of component (B), the daily intake is preferably 0.001 to 20 mg, more preferably 0.01 to 2 mg, even more preferably 0.1 to 0.7 mg, and particularly preferably 0.3 to 0.4 mg.

[0038] When the agent according to the present embodiment contains a polyamine composition as component (B), the daily intake amount of the polyamine composition will vary depending on the condition of the individual taking it (body weight, age, sex, etc.), the formulation, etc., but from the viewpoints of ease of ingestion as a single dose, the effectiveness of the physiological action based on component (B), safety, and minimizing the impact of the odor of component (B), the daily intake amount is preferably 50 to 400 mg, more preferably 100 to 300 mg, even more preferably 200 to 250 mg, and particularly preferably 201 to 220 mg.

[0039] Herbal medicines refer to natural products, such as plants, animals, and minerals, taken whole or in part for medicinal purposes, either as is or after simple processing such as drying. In the agent according to this embodiment, the form of the herbal medicine may include, for example, the herbal medicine itself (original herbal medicine); herbal powder obtained by drying and powdering the original herbal medicine; and herbal extracts obtained by extracting the original herbal medicine or herbal powder with water, an organic solvent such as ethanol, or a mixture of these solvents. Among these, herbal extracts are preferred from the viewpoint of achieving the effects of the present invention more significantly. The herbal extract may be in the form of a liquid extract (e.g., tincture, liquid extract), a diluted or concentrated extract (e.g., soft extract), or a dried extract that has been powdered or pasted (e.g., dried extract).

[0040] Specific examples of herbal extracts include ginger extract, maca extract, licorice extract, Astragalus extract, Tangerine Peony extract, peony extract, Panax ginseng extract, turmeric extract, Cistanche extract, cinnamon bark extract, Eleuthero extract, Hawthorn extract, Rehmannia root extract, Angelica acutiloba extract, Saw palmetto extract, Ginkgo leaf extract, Bupleurum Root extract, Cimicifuga Root extract, Dioscorea extract, Atractylodes Root extract, Atractylodes Root extract, Ganoderma lucidum extract, Rokujo extract, Tongkat Ali extract, French maritime pine bark extract, European oak extract, wild yam extract, diosgenin-containing yam extract, Cistanche Tubulosa extract, garlic extract, Eleuthero extract, Mucuna pruriens extract, Gardenia extract, Shilajit extract, Cordyceps mycelium powder, black ginger extract, black pepper extract, pomegranate seed extract, chaste tree extract, black cohosh extract, and dandelion extract. As the herbal extract, from the viewpoint of formulation stability, maca extract, Panax ginseng extract, saw palmetto extract, ginger extract, tongkat ali extract, French maritime pine bark extract, European oak extract, wild yam extract, diosgenin-containing yam extract, pomegranate seed extract, and chaste tree extract are preferred, with maca extract and Panax ginseng extract being more preferred.

[0041] Commercially available herbal medicines may be used. One herbal medicine may be used alone, or two or more herbal medicines may be used in combination.

[0042] The content of component (C) in the agent according to this embodiment is not particularly limited and is set appropriately depending on the type of component (C), the types and contents of other blended components, the dosage form, etc. From the viewpoint of more significantly exhibiting the effects of the present invention, the content of component (C) is, for example, preferably 0.1 to 70% by weight, more preferably 1 to 10% by weight, and even more preferably 2 to 4% by weight, based on the total amount of the agent according to this embodiment.

[0043] The content ratio of component (C) to component (A) in the agent according to this embodiment is not particularly limited and is set appropriately depending on the types of components (A) and (C), the types and contents of other blended components, the dosage form, etc. From the viewpoint of further enhancing the effects of the present invention, the content ratio of component (C) to component (A) is, for example, 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, of the total content of component (C) per 1 part by weight of the total content of component (A) contained in the agent according to this embodiment.

[0044] The content ratio of component (C) to component (B) in the agent according to this embodiment is not particularly limited and is set appropriately depending on the types of components (B) and (C), the types and contents of other blended components, the dosage form, etc. From the viewpoint of further enhancing the effects of the present invention, the content ratio of component (C) to component (B) is, for example, preferably 50 to 250 parts by weight, more preferably 80 to 200 parts by weight, and even more preferably 100 to 170 parts by weight, of the total content of component (C) per part by weight of the total content of component (B) contained in the agent according to this embodiment.

[0045] The agent according to this embodiment may contain a pharmacologically or physiologically active ingredient other than the ingredients (A), (B), and (C) as long as the effects of the present invention are not impaired. Specific examples of such pharmacologically or physiologically active ingredients include ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, folic acid, vitamin B12, vitamin C, vitamin D, vitamin A, vitamin E, niacin, calcium pantothenate, taurine, carnosine, anserine, balenine, citrulline, gamma-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, carnitine, glutamic acid, glutamine, and niacin. These include niacin, creatine, carnitine, luteolin, quercetin, genistin, 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), and equol. From the viewpoint of further enhancing the effects of the present invention, the pharmacologically active ingredient or physiologically active ingredient is preferably ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, resveratrol, isoflavone, equol, pycnogenol (proanthocyanidin), or folic acid, and more preferably ubiquinone (coenzyme Q10), astaxanthin, zinc, resveratrol, isoflavone, equol, pycnogenol (proanthocyanidin), or folic acid. One pharmacologically active ingredient or physiologically active ingredient may be used alone, or two or more may be used in combination.

[0046] The agent according to the present embodiment may contain various additives in addition to the above-mentioned components, provided that the effects of the present invention are not impaired. Specific examples of such additives include excipients, binders, disintegrants, lubricants, colorants, flavorings, odorants, sugars, sugar alcohols / polyalcohols, intense sweeteners, oils and fats, emulsifiers, thickeners, acidulants, and fruit juices. Examples of excipients include lactose, sucrose, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, and silicic acid. Examples of binders include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, calcium phosphate, and polyvinylpyrrolidone. Disintegrants include dry starch, sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, and lactose. Lubricants include talc, stearates, borax, and polyethylene glycol. Flavoring agents include sucrose, orange peel, citric acid, and tartaric acid. Sugars include sucrose, isomerized sugar, glucose, fructose, palatinose, trehalose, lactose, and xylose. Sugar alcohols and polyhydric alcohols include sorbitol, xylitol, erythritol, lactitol, palatinit, reduced starch syrup, reduced maltose syrup, glycerin, and propylene glycol. High-intensity sweeteners include aspartame, stevia, acesulfame potassium, and sucralose. Examples of fats and oils include safflower oil, grape seed oil, sunflower oil, olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, and perilla oil. Examples of emulsifiers include sucrose fatty acid esters, glycerin fatty acid esters, and lecithin. Examples of thickeners include carrageenan, xanthan gum, guar gum, pectin, and locust bean gum. Examples of acidulants include citric acid, lactic acid, and malic acid. Examples of fruit juices include lemon juice, orange juice, and berry juice.The additives may be used alone or in combination of two or more. From the viewpoint of the stability of the components (A) and (B) during formulation, the additive preferably contains an oil or fat.

[0047] The dosage form of the agent according to this embodiment is not particularly limited, and examples thereof include oral preparations such as tablets (including orally disintegrating tablets, chewable tablets, troche tablets, etc.), granules, powders, soft capsules, hard capsules, troches, jellies, or liquid preparations (including suspensions, emulsions, syrups, etc.); external preparations such as ointments, suppositories, patches, and sprays; injections, etc. Among these, from the viewpoint of ease of handling and further enhancing the effects of the present invention, oral preparations are preferred, and tablets, granules, soft capsules, and hard capsules are more preferred.

[0048] The agent according to the present embodiment can be used as an ingredient of, for example, pharmaceuticals, quasi-drugs, cosmetics, and food and beverages (drinks, foods). The agent according to the present embodiment can also be used as, for example, pharmaceutical preparations, quasi-drug preparations, foods for specified health uses, foods with nutrient function claims, foods for the elderly, foods for special dietary uses, foods with functional claims, health supplements (supplements), food preparations (e.g., confectionery tablets), and clear foods. Furthermore, the agent according to the present embodiment can also be used as, for example, veterinary medicines and feed additives.

[0049] When the agent according to the present embodiment is used as a food, the food may be a general food compounded with component (A) and, if necessary, other ingredients, such as solid foods such as cookies, biscuits, snacks, jellies, gummies, chocolates, chewing gum, candies, and cheese; and liquid foods such as nutritional drinks, juices, tea drinks, coffee drinks, and dairy drinks.

[0050] The agent according to this embodiment can be suitably used in subjects requiring an effect of suppressing the progression of aging of female reproductive tissues (e.g., humans; non-human animals such as cattle, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets). From the viewpoint of further enhancing the effects of the present invention, humans are preferred as subjects. Furthermore, from the viewpoint of further enhancing the effects of the present invention, among non-human animals, cattle, pigs, and horses are preferred, and pigs are more preferred.

[0051] 2. Method for inhibiting the progression of aging of ovaries, follicles, or eggs Pyrroloquinoline quinone or a salt thereof has the effect of suppressing the progression of aging in ovaries, follicles, or eggs. Therefore, one embodiment of the present invention provides a method for suppressing the progression of aging in ovaries, follicles, or eggs, which comprises ingesting a composition containing pyrroloquinoline quinone or a salt thereof. The ovaries, follicles, or eggs may be human ovaries, follicles, or eggs, or may be ovaries, follicles, or eggs of non-human animals such as cows, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets. From the viewpoint of further enhancing the effects of the present invention, human ovaries, follicles, or eggs are preferred. Furthermore, from the viewpoint of further enhancing the effects of the present invention, among the ovaries, follicles, or eggs of non-human animals, bovine, porcine, or equine ovaries, follicles, or eggs are preferred, and porcine ovaries, follicles, or eggs are more preferred.

[0052] In these embodiments, the type, content, and daily intake amount of pyrroloquinoline quinone or a salt thereof in the composition, the type, content, and daily intake amount of other components, and the formulation form of the composition are as described in [1. Anti-aging agent for ovaries, follicles, or ova]. [Example]

[0053] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to the following examples. All of the following test examples were conducted with the approval of the Hiroshima University Animal Experiment Ethics Committee.

[0054] [Test Example 1: Effect of administration of pyrroloquinoline quinone on blood AMH (anti-Müllerian hormone) in female mice] Six-month-old female Leydig cell-specific NRG1-deficient mice were administered PQQ disodium salt (2 mg / kg / day) in their drinking water for two weeks (n = 5). Six-month-old Leydig cell-specific NRG1-deficient male mice (KO; control) and 6-month-old wild-type mice (WT) were also administered water alone for two weeks. Blood samples were then collected, and the concentration of AMH (anti-Müllerian hormone), a follicular marker, was measured using an AMH ELISA kit (Elabscience Biotechnology, Bethesda, MD) according to the kit's instructions. The results are shown in Figure 1. Leydig cell-specific NRG1-deficient female mice have a shorter lifespan; 6 months of age corresponds to a human age in their 40s.

[0055] As shown in Figure 1, the AMH concentration was reduced in control female mice not administered PQQ disodium salt compared to WT mice, whereas the AMH concentration was increased in female mice administered PQQ disodium salt compared to WT mice. This indicates that intake of PQQ disodium salt rejuvenates ovarian tissue.

[0056] [Test Example 2: Effect of administration of pyrroloquinoline quinone on the estrous cycle in female mice] Six-month-old Leydig cell-specific NRG1-deficient female mice (KO) were continuously administered PQQ disodium salt in their drinking water (2 mg / kg / day). Female mice receiving only water served as controls. The estrous cycle of each mouse was assessed by vaginal smear examination. The vaginal smear examination was performed as follows: saline was perfused into the vagina, and the perfused material was smeared onto a glass slide. After drying, the smear was fixed with methanol and stained with Giemsa. The smears were observed under an inverted microscope (Keyence). The presence of only leukocytes indicated diestrus; the presence of only nucleated epithelial cells indicated proestrus; the presence of only anucleated epithelial cells indicated estrus; the presence of a mixture of anucleated epithelial cells and leukocytes, with a greater number of anucleated epithelial cells; and the presence of a mixture of anucleated epithelial cells and leukocytes, with a greater number of leukocytes, indicated metestrus. The results are shown in Figure 2.

[0057] As shown in Figure 2, control female mice not administered PQQ disodium salt maintained a constant microestrus state and showed no cyclical changes in estrus, whereas in female mice administered PQQ disodium salt, cyclical changes in estrus were confirmed to resume around 14 days after the start of administration. In other words, it was confirmed that administration of PQQ disodium salt improved the abnormal ovulation cycle in female mice.

[0058] [Test Example 3: Effect of administration of pyrroloquinoline quinone on eggs] Three-week-old immature female mice were administered PQQ disodium salt in drinking water (2 mg / kg / day) (n = 5). Female mice administered only water served as controls. Two days after the start of administration, 4 IU of eCG (pregnant mare serum gonadotropin) was administered to induce follicular development, followed 48 hours later by 5 IU of hCG (human chorionic gonadotropin). The number of ovulations in each female mouse was then measured. Oviducts were removed from sacrificed mice, the ampulla of the oviduct was dissected, and the ovulated cumulus cell complexes were collected. The complexes were then visually observed under a stereomicroscope. In vitro fertilization was performed using the ovulated oocytes, and the fertilization rate and development rate were calculated. Here, fertilized oocytes refer to cleaved oocytes 24 hours after fertilization, and the fertilization rate refers to the ratio of successfully fertilized oocytes to the total number of mature oocytes ovulated. The development rate refers to the proportion of fertilized eggs that develop into blastocyst-stage embryos. The results of the number of ovulations and fertilization rate, expressed as mean ± standard deviation, are shown in Table 1. Note that 3 weeks of age in mice corresponds to the teenage years when menstruation begins in humans.

[0059] [Table 1]

[0060] As shown in Table 1, female mice administered PQQ disodium salt showed an increased number of ovulations while maintaining the in vitro fertilization rate, compared with control female mice not administered PQQ disodium salt, and also showed an improved development rate while maintaining the in vitro fertilization rate. In other words, it was confirmed that administration of PQQ disodium salt improves the quality of eggs.

[0061] [Test Example 4: Effect of administration of pyrroloquinoline quinone on blood female hormone concentrations] Three-week-old immature female mice were administered PQQ disodium salt in drinking water (2 mg / kg / day). Female mice administered only water served as controls. Two days after the start of administration, 4 IU of eCG (pregnant mare serum gonadotropin) to induce follicular development was intraperitoneally administered, followed 48 hours later by 5 IU of hCG (human chorionic gonadotropin). Serum was collected from each female mouse over time, beginning immediately before eCG administration, and the concentrations of estradiol (E2), a marker of follicular development, and progesterone (P4), a marker of corpus luteum, were measured using commercially available kits from Endocrine Technology, Inc. (Newark, CA). The results are shown in Figure 3.

[0062] As shown in Figure 3(A), blood estradiol concentrations increased over time in female mice administered PQQ disodium salt compared with control female mice not administered PQQ disodium salt. This confirms that administration of PQQ disodium salt promotes estradiol production. Estradiol is a female hormone widely used in hormone replacement therapy and is widely known to be used to treat menopausal disorders and alleviate menopausal symptoms. This confirms that pyrroloquinoline quinone or its salts are useful as an agent for improving or preventing symptoms resulting from estrogen deficiency in females, as well as for improving or preventing premenopausal or menopausal disorders in females. This allows premenopausal or menopausal women to experience menopause without pain. Furthermore, as shown in Figure 3(B), there was no significant difference in the increase in blood progesterone concentration between female mice administered PQQ disodium salt and control female mice not administered PQQ disodium salt. Progesterone is known to be involved in blood concentration in ovarian hyperstimulation syndrome (OVHS). In other words, pyrroloquinoline quinone or its salts were confirmed to be useful as a preventive agent for OVHS by promoting estradiol production while mitigating progesterone production.

[0063] [Test Example 5: Effect of administration of pyrroloquinoline quinone on ovarian fibrosis] Six-month-old Leydig cell-specific NRG1-deficient female mice (KO) were administered PQQ disodium salt in their drinking water (2 mg / kg / day) for two weeks. The ovaries were then excised, stained with PSR, and observed under an inverted microscope (Keyence) at 20x magnification. The results are shown in Figure 4.

[0064] As shown in Figure 4, in mice before administration of PQQ disodium salt, the interstitial cells were stained red throughout, confirming collagen deposition, i.e., ovarian fibrosis. At the same time, atrophy of interstitial follicles was also observed. On the other hand, in mice administered PQQ disodium salt for two weeks, the red color of the interstitial cells disappeared, confirming that the fibrotic interstitial cells had returned to a healthy state. Ovarian interstitial cells are known to become increasingly fibrotic with age. In other words, it was confirmed that the intake of PQQ disodium salt rejuvenated ovarian tissue. Furthermore, antral follicles were observed, and follicular atrophy was also improved, confirming that egg quality had improved.

[0065] The results of Test Examples 1 to 5 indicate that taking pyrroloquinoline quinone or a salt thereof can maintain women's quality of life, which tends to be impaired by physical and mental abnormalities that occur as menopause approaches, and can help women stay vibrant and youthful even at an older age.

Claims

[Claim 1] An agent for increasing the number of ovulated eggs for use in a human or a non-human animal, comprising pyrroloquinoline quinone or a salt thereof, wherein the non-human animal is a pig, horse, sheep, goat, dog, rabbit, mouse, rat, guinea pig, gerbil, hamster, or ferret.

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