WO2020262325A1

WO2020262325A1 - Anti-aging agent for females

Info

Publication number: WO2020262325A1
Application number: PCT/JP2020/024469
Authority: WO
Inventors: 昌之島田
Original assignee: 国立大学法人広島大学; ロート製薬株式会社
Priority date: 2019-06-28
Filing date: 2020-06-22
Publication date: 2020-12-30
Also published as: TW202114674A; CN114007618A; JPWO2020262325A1

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

Female anti-aging agent

The present invention relates to a female anti-aging agent.

In women, the aging of reproductive tissues such as ovaries, follicles, and eggs progresses with aging, making pregnancy and childbirth difficult. In fertility treatment of such women who have difficulty in pregnancy, hormone replacement therapy is often used to induce ovulation. However, it is difficult to develop and ovulate uniformly mature follicles, and as a side effect, a symptom called ovarian hyperstimulation syndrome (OHSS) is exhibited, and if this worsens, there is a risk of respiratory failure or circulatory failure. Therefore, in fertility treatment, a safe and effective treatment method is required (Non-Patent Document 1).

In addition, when the concentration of female hormones such as estrogen decreases with the progress of aging of reproductive tissues in women, menopause is mainly caused in women in their late 40s to 50s. In some cases, even women in their 30s to mid-40s, combined with stress and overwork, may exhibit irregular menstruation in addition to the above-mentioned symptoms of menopause. This is also called pre-menopause, and along with menopause, it significantly reduces women's quality of life. Hormone replacement therapy is mainly used in the treatment of these menopausal disorders and pre-menopausal disorders, but there are problems of side effects such as the possibility of carcinogenesis, irregular bleeding, vaginal bleeding, headache, and liver dysfunction. While pre-menopause and menopause can occur in any woman, it is very important for women to spend their time from pre-menopause to menopause, especially in Japan, where the population is aging. is there. Therefore, as with infertility treatment, a safe and effective treatment method for pre-menopausal disorders and menopausal disorders is required (Non-Patent Document 2).

By the way, aging of female reproductive tissues has become a problem not only in humans but also in domestic animals such as cattle and pigs. For example, in the pig farming industry, the cycle from mating to calving of sows is 2.5 times a year, but the number of offspring of sows decreases with aging and breeding due to an increase in reproductive disorders such as miscarriage. Grades also deteriorate. Therefore, it is desired to develop a method for suppressing the progress of aging of female reproductive tissues not only in humans but also in non-human animals.

An object of the present invention is to provide an anti-aging agent for an ovary, a follicle or an egg (hereinafter, these may be collectively referred to as "female reproductive tissue").

The present inventors have found that pyrroloquinoline quinone or a salt thereof suppresses the progress of aging of female reproductive tissues and promotes the production of estradiol. The present invention is based on this novel finding.

The present invention provides, for example, the following inventions.
[1]
An ovarian, follicle or egg anti-aging agent containing pyrroloquinoline quinone or a salt thereof.
[2]
An inhibitor or ameliorating agent for fibrosis of female reproductive tissues, which contains pyrroloquinoline quinone or a salt thereof.
[3]
An egg quality improver containing pyrroloquinoline quinone or a salt thereof.
[4]
An improving or preventing agent for menstrual irregularities containing pyrroloquinoline quinone or a salt thereof.
[5]
An ameliorating or prophylactic agent for ovarian hyperstimulation syndrome containing pyrroloquinoline quinone or a salt thereof.
[6]
An estrogen production promoter containing pyrroloquinoline quinone or a salt thereof.
[7]
The agent according to [6], which ameliorates or prevents symptoms caused by female estrogen deficiency.
[8]
The agent according to [6], which improves or prevents pre-menopause or menopause in females.
[9]
The agent according to [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 of suppressing the progress of aging of an ovary, a follicle or an egg by ingesting a composition containing pyrroloquinoline quinone or a salt thereof.
[2-1]
The agent according to [1], which suppresses or improves fibrosis of female reproductive tissue.
[2-2]
The agent according to [1], which improves the quality of eggs.
[2-3]
The agent according to [1], which improves or prevents menstrual irregularities.
[2-4]
The agent according to [1], which improves or prevents ovarian hyperstimulation syndrome.
[2-5]
The agent according to [1], which promotes the production of estrogen.
[2-6]
The agent according to [2-5], which improves or prevents symptoms caused by female estrogen deficiency.
[2-7]
The agent according to [2-5], which improves or prevents pre-menopausal disorder or climacteric disorder in females.
[2-8]
The agent according to [2-5], which improves or prevents menopausal symptoms in females.

According to the present invention, it is possible to provide an anti-aging agent for female reproductive tissues.

In Test Example 1, it is a graph which shows the blood AMH concentration in each female mouse. (A) is a graph showing the estrous cycle when pyrroloquinoline quinone disodium salt is not administered to female aging model mice in Test Example 2. (B) is a graph showing the estrous cycle when pyrroloquinoline quinone was administered to a female aging model mouse in Test Example 2. In FIG. 2, M indicates the late estrous cycle, WE indicates the microestrous cycle, E indicates the estrous cycle, P indicates the early estrous cycle, and D indicates the telogen effluvium. (A) shows the time and blood estradiol concentration when eCG (equine chorionic gonadotropin) was administered to immature female mice to which pyrroloquinoline quinone disodium salt was administered or not administered in Test Example 4. It is a graph which shows the relationship. In Test Example 4, (B) administered eCG (equine chorionic gonadotropin) and hCG (human chorionic gonadotropin) to immature female mice to which pyroloquinolinquinone disodium salt was administered or not. It is a graph which shows the relationship between time and blood progesterone concentration at time. (A) is a photograph of the ovary of a female aging model mouse before administration of pyrroloquinoline quinone disodium salt in Test Example 5 taken by PSR staining (magnification 20 times). (B) is a photograph of the ovary after administration of pyrroloquinoline quinone disodium salt for 2 weeks to a female aging model mouse in Test Example 5 by PSR staining (magnification 20 times). (C) and (D) are traces of (A) and (B), respectively, and shaded areas indicate stained areas.

Hereinafter, a mode for carrying out the present invention will be described in detail. However, the present invention is not limited to the following embodiments.

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

Pyrroloquinoline quinone or a salt thereof has an effect of suppressing the disappearance or decrease of AMH (Anti-Mullerian Hormone; anti-Müllerian hormone) in females. Here, AMH is a hormone that is an index of aging of female reproductive tissues (particularly ovaries), and it is known that the blood AMH concentration decreases as the aging of female reproductive tissues progresses. In addition, pyrroloquinoline quinone or a salt thereof has an effect of suppressing or improving fibrosis of female reproductive tissues (ovary, follicle, egg, etc., particularly ovary). Therefore, as an embodiment of the present invention, an anti-aging agent for ovary, follicle or egg, and an agent for suppressing or improving fibrosis of female reproductive tissue, which contains pyrroloquinoline quinone or a salt thereof, are provided. "Anti-aging" here means rejuvenating women, suppressing the progress of aging of women, being able to maintain a high QOL even when women are old, and being lively and young even when women are old. It refers to being able to spend time in a different way. Further, the term "elderly" as used herein refers to those in their thirties or older, preferably in their forties or older, and more preferably in their fifties or older. An example of aging of female reproductive tissue (particularly ovarian tissue) is ovarian reserve (the number of secondary follicles remaining in the ovary). As described above, since pyrroloquinoline quinone or a salt thereof has an effect of suppressing the disappearance or decrease of AMH in females, it also has an effect of improving ovarian reserve.

Pyrroloquinoline quinone or its salt has the effect of improving the quality of eggs. Therefore, as an embodiment of the present invention, an egg quality improving agent containing pyrroloquinoline quinone or a salt thereof is provided. Here, "improvement of egg quality" refers to maintaining or improving the fertilization rate of the ovulated egg while increasing the number of ovulations, and the fertilization rate here refers to fertilization with respect to the total number of ovulated eggs. It is the ratio of the number of ovulated eggs. Therefore, as an embodiment of the present invention, there is provided an agent containing pyrroloquinoline quinone or a salt thereof for increasing the number of ovulated eggs and maintaining or improving the fertilization rate of the ovulated eggs. .. In addition, "improvement of egg quality" refers to maintaining or improving the development rate while maintaining or improving the fertilization rate of the egg, and the development rate here means the blastocyst stage embryo of the fertilized egg. Refers to the proportion of those that became. Therefore, as an embodiment of the present invention, an agent containing pyrroloquinoline quinone or a salt thereof for maintaining or improving the fertilization rate of an egg and maintaining or improving the incidence rate is provided.

Pyrroloquinoline quinone or its salt has the effect of improving or preventing menstrual irregularities. Therefore, as an embodiment of the present invention, an improving agent or a preventive agent for irregular menstruation containing pyrroloquinoline quinone or a salt thereof is provided. Here, "irregular menstruation" means a state in which the cycle of menstruation (ovulation in female non-human animals without menstruation) in females is abnormal. For example, in humans, the normal menstrual cycle is 25 to 38 days, and the menstrual cycle is within 24 days or 39 days or more, or there is no menstruation for 3 months or more (in the case of amenorrhea). Menstruation is irregular.

Pyrroloquinoline quinone or a salt thereof has an effect of promoting the production of estrogen (estron, estradiol, estriol, preferably estradiol). Therefore, as an embodiment of the present invention, an estrogen production promoter containing pyrroloquinoline quinone or a salt thereof is provided. In addition, pyroloquinolinquinone or a salt thereof promotes the production of estrogen, thereby improving or preventing the symptoms caused by estrogen deficiency in females, improving or preventing menopausal symptoms in females, and pre-menopausal disorders in females. In addition to having the effect of improving or preventing menopause or improving or preventing menopause in females, it also has the effect of supplementing female hormones. Therefore, as one embodiment of the present invention, a symptom ameliorating agent or a preventive agent for female estrogen deficiency, a female climacteric symptom ameliorating agent or a preventive agent, or a female pre. Menopausal disorders or ameliorating or prophylactic agents for menopause are provided.

Pyrroloquinoline quinone or a salt thereof has the effect of improving or preventing ovarian hyperstimulation syndrome. Therefore, as an embodiment of the present invention, an ameliorating agent or a preventive agent for ovarian hyperstimulation syndrome containing pyrroloquinoline quinone or a salt thereof is provided. Here, "ovarian hyperstimulation syndrome" means various symptoms caused by ascites and pleural effusion caused by swelling of the ovary due to excessive stimulation of an ovarian inducer, and in severe cases, blood is concentrated and renal failure or It also causes complications such as thrombosis. Specific symptoms of ovarian hyperstimulation syndrome include bloating, nausea, rapid weight gain, decreased urine output, abdominal pain, diarrhea and the like. In addition, pyrroloquinoline quinone or a salt thereof has an effect of improving or preventing ovarian hyperstimulation syndrome, thereby improving or preventing symptoms derived from ovarian hyperstimulation syndrome and complications of ovarian hyperstimulation syndrome. In addition, the agent containing pyrroloquinoline quinone or a salt thereof according to each of the above embodiments is also collectively referred to as "agent according to this embodiment".

In this specification, "menopause" means about 10 years before and after menopause in females, and in the case of humans, the period from the late 40s to the 50s corresponds to the menopause. Further, in the present specification, the term "pre-menopause" refers to a period younger than the above-mentioned menopause, in which symptoms of pre-menopause described later may be exhibited, and in the case of humans, the period from the 30s to the mid-40s is. Corresponds to pre-menopause.

Symptoms of "female menopause" include, for example, suffocation, lightheadedness, dizziness, numbness in limbs, stiffness in limbs, swelling, tiredness, ringing in the ears, swelling, hot flash, sweating, palpitation, excitement, etc. Sleeplessness, irritation, anger, emotional instability, depressed mood, fragile tears, decreased motivation, anxiety, headache, stiff shoulders, chest pain, lower back pain, joint pain, cold hands and feet, itching, dry skin and eyes, nausea , Loss of appetite, abdominal pain, constipation, dizziness, frequent urination, palpitations, palpitations, etc.

"Symptoms derived from female estrogen deficiency" include, for example, menopause, autonomic imbalance, neuropsychiatric symptoms, urogenital atrophy, hyperlipidemia, arteriosclerosis, hypertension, stroke, osteoporosis, bone mass. In addition to the symptoms of osteoporosis, skin atrophy, joint pain, dry mouth, taste changes, sleep disorders, and the above-mentioned "female menopause", the "Hormone Replacement Therapy Guidelines 2017" edited by the Japan Society of Obstetrics and Gynecology Symptoms and the like described in "1. Expected actions / effects of HRT" can be mentioned.

Symptoms of "female pre-menopause (female juvenile menopause)" include irregular menstruation, amenorrhea, etc., in addition to the above-mentioned "female menopause" symptoms.

"Female menopause" refers to the above-mentioned "female menopause" symptoms and "female pre-menopause (female juvenile menopause)" symptoms. However, the symptom can be exhibited not only from "menopause" or "pre-menopause" but also from a younger age (for example, in the case of a human being in his teens when menstruation begins). As the "female menopausal symptom", among these specific symptom described above, the symptom may be mild. For example, there may be a state in which the person is aware of burning or sweating but the number of times is small, or a state in which the degree of depression or stiff shoulder is small.

Pyrroloquinoline quinone has the following formula:

It is a known compound represented by.

Examples of the pyroloquinolinquinone salt include alkali metal salts, alkaline earth metal salts and ammonium salts. Examples of the alkali metal salt include sodium salt, potassium salt and lithium salt. Examples of the alkaline earth metal salt include calcium salt and magnesium salt.

As the pyrroloquinoline quinone or a salt thereof, an alkali metal salt of pyrroloquinoline quinone is preferable, a sodium salt of pyrroloquinoline quinone is more preferable, and a disodium salt of pyrroloquinoline quinone is further preferable.

As the pyrroloquinoline quinone or a salt thereof, commercially available ones can also be used. Further, as the pyrroloquinoline quinone or a salt thereof, for example, natto, soybean, cocoa powder, cacao mass, cacao, parsley, peppers and the like containing pyrroloquinoline quinone or a salt thereof may be used.

When used in humans, the content of component (A) in the agent according to this embodiment is the total amount of component (A) based on the total amount of the agent according to this embodiment from the viewpoint of the stability of the drug to be administered. The content is preferably 0.01 to 30% by weight, more preferably 0.2 to 10% by weight, further preferably 1 to 5% by weight, and 1.3 to 3% by weight. Is particularly preferable. When used in non-human animals, the content of the component (A) in the agent according to the present embodiment is such that the total content of the component (A) is 0, based on the total amount of the agent according to the present embodiment. It is preferably 01 to 0.3% by weight, more preferably 0.02 to 0.2% by weight.

In the agent according to the present embodiment, the daily intake of the component (A) may differ depending on the condition (body weight, age, sex, etc.) of the individual ingesting, the formulation form, etc., but when used in humans, From the viewpoint of ease of ingestion as a single dose, the viewpoint of the effectiveness of the physiological action based on the component (A), and the viewpoint of safety, preferably 7 to 100 mg, more preferably 10 to 40 mg, still more preferably 20 to 20 to It is 25 mg, and when used for non-human animals, it is preferably 1 to 30 mg / kg, more preferably 2 to 20 mg / kg.

The agent according to the present embodiment may further contain a polyamine (also referred to as "component (B)") and a crude drug (also referred to as "component (C)"). As a result, the effect of the present invention is more remarkable.

Polyamine is an aliphatic hydrocarbon containing two or more primary amino groups in the molecule. Specific examples of polyamines include, for example, ptolessin, cadaverine, ethylenediamine, trimethylenediamine, hexamethylenediamine, spermine, kardin, homospermine, aminopropyl cadaverine, spermine, thermin, thermospermine, canabalmin, aminopentylnorspermine, aminopropyl. Homo spermine, canabalmin, homospermine, cardopentamine, homocardopentamine, aminopropyl canabalmin, bis (aminopropyl) homospermine, bis (aminopropyl) norspermine, aminobutyl canabalmin, aminopropyl homospermine, Examples thereof include homopentamine, cardohexamine, homocardohexamine, selmohexamine, and homoselmohexamine. As the polyamine, spermidine, spermine, and putrescine are preferable, spermidine and spermine are more preferable, and spermidine is further preferable, from the viewpoint of ease of use.

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. Further, as polyamines, extracts from plants such as soybean (preferably soybean germ), wheat (preferably wheat germ) or rice (preferably rice germ), extracts from seafood (preferably shirako), and dried sake. A composition containing a polyamine such as yeast (also referred to as a "polyamine composition") can also be used. When the polyamine composition is an extract from a plant, water, an organic solvent such as ethanol, or a mixed solvent thereof can be used as the extraction solvent. As the polyamine composition, an extract from a plant is preferable, and an extract from soybean (preferably soybean germ) is more preferable from the viewpoint of ease of use. Examples of commercially available polyamine compositions include "Soipolya" (manufactured by Combi Co., Ltd.), "Oryzapolyamine-P" (manufactured by Oryza Yuka Co., Ltd.), "Oryzapolyamine-LC" (manufactured by Oryza Yuka Co., Ltd.) Examples thereof include "Fight Polyamine-S" (manufactured by Toyobo Co., Ltd.), "Fight Polyamine-SP" (manufactured by Toyobo Co., Ltd.), and "Elion SP" (manufactured by Mitsubishi Gas Chemicals Co., Ltd.).

The content of the component (B) in the agent according to the present embodiment is not particularly limited, and is appropriately set according to the type of the component (B), the type and content of other compounding components, the formulation form, and the like. The content of the component (B) is determined based on, for example, the total amount of the agent according to the present embodiment from the viewpoint of the stability of the preparation and the viewpoint of reducing the influence of the odor of the component (B). The total content of is preferably 0.001 to 10% by weight, more preferably 0.01 to 0.1% by weight, still more preferably 0.02 to 0.05% by weight. ..

When the agent according to the present embodiment contains the polyamine composition as the component (B), the content of the polyamine composition is not particularly limited, and the type of the polyamine composition, the type and content of other compounding components, and the preparation. It is appropriately set according to the form and the like. The content of the polyamine composition is determined from the viewpoint of the stability of the preparation, the viewpoint of the effectiveness of the physiological action based on the component (B), the viewpoint of safety, and the viewpoint of reducing the influence of the odor of the component (B). For example, based on the total amount of the agent according to the present embodiment, the total content of the polyamine composition is preferably 5 to 35% by weight, more preferably 10 to 30% by weight, and 14 to 23% by weight. It is more preferably%.

The content ratio of the component (B) to the component (A) in the agent according to the present embodiment is not particularly limited, and the types of the components (A) and (B), the types and contents of other compounding components, and the formulation form. It is set appropriately according to the above. Regarding the content ratio of the component (B) to the component (A), from the viewpoint of further enhancing the effect of the present invention, for example, with respect to 1 part by weight of the total content of the component (A) contained in the agent according to the present embodiment. The total content of the component (B) is preferably 0.001 to 0.1 parts by weight, more preferably 0.01 to 0.03 parts by weight, and 0.015 to 0.02. It is more preferably a part by weight.

When the agent according to the present embodiment contains the polyamine composition as the component (B), the content ratio of the polyamine composition to the component (A) is not particularly limited, and the type of the component (A) and the polyamine composition, etc. It is appropriately set according to the type and content of the compounding components of the above, the formulation form, and the like. Regarding the content ratio of the polyamine composition to the component (A), from the viewpoint of further enhancing the effect of the present invention, for example, with respect to 1 part by weight of the total content of the component (A) contained in the agent according to the present embodiment. The total content of the polyamine composition is preferably 5 to 15 parts by weight, more preferably 7 to 10 parts by weight.

In the agent according to the present embodiment, when the component (B) contains spermidine and spermine, the weight ratio of spermine to spermidine is not particularly limited, and the types of the components (A) and (B) and other compounding components. It is appropriately set according to the type, content, formulation form and the like. The weight ratio of spermine to spermidine is preferably, for example, 0.01 to 10, more preferably 0.1 to 2, and 0.2 to 0 from the viewpoint of further enhancing the effect of the present invention. It is more preferably .5.

In the agent according to the present embodiment, the daily intake of the component (B) may differ depending on the condition (body weight, age, sex, etc.) of the individual ingesting, the formulation form, etc., but is based on the component (B). From the viewpoint of the effectiveness of physiological action, the viewpoint of safety, and the viewpoint of reducing the influence of the odor of the component (B), it is preferably 0.001 to 20 mg, more preferably 0.01 to 2 mg, and further preferably 0. It is 1 to 0.7 mg, particularly preferably 0.3 to 0.4 mg.

When the polyamine composition is contained as the component (B) in the agent according to the present embodiment, the daily intake of the polyamine composition depends on the condition (body weight, age, sex, etc.) of the individual ingesting, the formulation form, etc. It may vary depending on the viewpoint, but from the viewpoint of ease of ingestion as a single dose, from the viewpoint of (B) effectiveness of physiological action based on the component, from the viewpoint of safety, and from the viewpoint of reducing the influence of the odor of the component (B). Therefore, it is preferably 50 to 400 mg, more preferably 100 to 300 mg, still more preferably 200 to 250 mg, and particularly preferably 201 to 220 mg.

Crude drugs are those that have undergone simple processing such as drying all or part of natural products such as plants, animals, and minerals, or drying them, for the purpose of medicinal purposes. In the agent according to the present embodiment, as the form of the crude drug, for example, the crude drug itself (herbal medicine); the crude drug powder obtained by drying and powdering the crude drug; the crude drug or the crude drug powder in an organic solvent such as water or ethanol, or these. Examples thereof include crude drug extracts extracted with a mixed solvent of. Among these, crude drug extracts are preferable from the viewpoint of exerting the effect of the present invention more remarkably. The crude drug extract may be the extract as it is (tincture, stream extract, etc.), may be a diluted or concentrated extract (soft extract, etc.), or the extract may be dried and then pulverized. Alternatively, it may be in the form of a paste (dried extract, etc.).

Specific examples of the crude drug extract include ginger extract, maca extract, kanzo extract, ougi extract, chimpi extract, shakyaku extract, otane carrot extract, corn extract, nikjuyo extract, keihi extract, elephant kogi extract, sanzashi extract, dioscorea extract, and touki extract. , Nokogiri palm extract, Ginkgo biloba extract, Psycho extract, Ginger extract, Sanyaku extract, Taisou extract, Byakujutsu extract, Sojutsu extract, Reishi extract, Rokujo extract, Tonkat Ali extract, French coastal pine bark extract, European oak extract, Wild yam Extract, diosgenin-containing mountain potato extract, canca extract, garlic extract, eleuthero extract, mukuna extract, cuttlefish extract, shiragit extract, winter worm summer grass mycelium powder, black ginger extract, black pepper extract, pomegranate seed extract, chest tree extract, black kohosh extract, western Examples include dandelion extract. From the viewpoint of formulation stability, crude drug extracts include maca extract, otane carrot extract, sawtooth palm extract, ginger extract, toncat ant extract, French coastal pine bark extract, European oak extract, wild yam extract, diosgenin-containing yam extract, and pomegranate seeds. Extracts and chest tree extracts are preferable, and maca extract and otane carrot extract are more preferable.

As the crude drug, commercially available ones may be used. As the crude drug, one kind may be used alone, or two or more kinds may be used in combination.

The content of the component (C) in the agent according to the present embodiment is not particularly limited, and is appropriately set according to the type of the component (C), the type and content of other compounding components, the formulation form, and the like. Regarding the content of the component (C), from the viewpoint of exerting the effect of the present invention more remarkably, for example, the total content of the component (C) is 0.1 to 0 based on the total amount of the agent according to the present embodiment. It is preferably 70% by weight, more preferably 1 to 10% by weight, and even more preferably 2 to 4% by weight.

The content ratio of the component (C) to the component (A) in the agent according to the present embodiment is not particularly limited, and the types of the components (A) and (C), the types and contents of other compounding components, and the formulation form. It is set appropriately according to the above. Regarding the content ratio of the component (C) to the component (A), from the viewpoint of further enhancing the effect of the present invention, for example, with respect to 1 part by weight of the total content of the component (A) contained in the agent according to the present embodiment. The total content of the component (C) is preferably 0.1 to 3 parts by weight, more preferably 0.5 to 2.5 parts by weight, and 1 to 1.5 parts by weight. Is even more preferable.

The content ratio of the component (C) to the component (B) in the agent according to the present embodiment is not particularly limited, and the types of the component (B) and the component (C), the types and contents of other compounding components, and the formulation form. It is set appropriately according to the above. Regarding the content ratio of the component (C) to the component (B), from the viewpoint of further enhancing the effect of the present invention, for example, with respect to 1 part by weight of the total content of the component (B) contained in the agent according to the present embodiment. The total content of the component (C) is 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.

The agent according to the present embodiment may contain a pharmacologically active ingredient or a physiologically active ingredient other than the components (A), (B) and (C) as long as the effects of the present invention are not impaired. Specific examples of such pharmacologically active ingredients or physiologically active ingredients include ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, folic acid, vitamin B12, vitamin C, vitamin D, vitamin A, and vitamin E. Niacin, calcium pantothenate, taurine, carnosin, anserine, valenin, citrulin, γ-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, carnitine, glutamic acid, glutamine, creatine, carnitine, luteolin, quercetin, genistin, cyanidin, Resveratrol, diosgenin, isoflavone aglycon, isoflavone, lipoic acid, zinc, iron, calcium, selenium, astaxanthin, zeaxanthin, β-carnitine, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, Placenta extract, Pycnogenol (proanthocyanidin), equol and the like can be mentioned. As the pharmacologically active ingredient or the physiologically active ingredient, ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, resveratrol, isoflavone, equol, from the viewpoint of further enhancing the effect of the present invention. Pycnogenol (proanthocyanidin) and folic acid are preferable, and ubiquinone (coenzyme Q10), astaxanthin, zinc, resveratrol, isoflavone, equol, pycnogenol (proanthocyanidin) and folic acid are more preferable. As the pharmacologically active ingredient or the physiologically active ingredient, one kind may be used alone, or two or more kinds may be used in combination.

The agent according to the present embodiment may contain various additives in addition to the above components as long as the effects of the present invention are not impaired. Specific examples of such additives include excipients, binders, disintegrants, thickeners, colorants, flavoring agents, flavoring agents, sugars, sugar alcohols / polyhydric alcohols, and high-sweetness sweetness. Materials, fats and oils, emulsifiers, thickeners, acidulants, fruit juices and the like can be mentioned. Examples of excipients include lactose, sucrose, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, and silicic acid. Examples of the binder include water, ethanol, propanol, simple 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 mentioned. Examples of the disintegrant include dried starch, sodium alginate, canten powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, lactose and the like. Examples of the lubricant include talc, stearate, borax, polyethylene glycol and the like. Examples of the flavoring agent include white sugar, orange peel, citric acid, tartaric acid and the like. Examples of sugars include sugars such as sucrose, high fructose corn syrup, glucose, fructose, palatinose, trehalose, lactose and xylose. Examples of sugar alcohols and polyhydric alcohols include sorbitol, xylitol, erythritol, lactitol, palatinit, reduced starch syrup, reduced malt sugar starch syrup, glycerin, propylene glycol and the like. Examples of high-sweetness sweeteners include aspartame, stevia, acesulfame potassium, sucralose and the like. Examples of fats and oils include safflower oil (safflower oil), grape seed oil, sunflower oil (sunflower oil), olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, and perilla oil. Examples of the emulsifier include sucrose fatty acid ester, glycerin fatty acid ester, lecithin and the like. Examples of the thickener include carrageenan, xanthan gum, guar gum, pectin, locust bean gum and the like. Examples of the acidulant include citric acid, lactic acid, malic acid and the like. Examples of fruit juices include lemon juice, orange juice, and berry juice. As the additive, one type may be used alone, or two or more types may be used in combination. The additive preferably contains fats and oils from the viewpoint of stability of the component (A) and the component (B) at the time of formulation.

The dosage form of the agent according to the present embodiment is not particularly limited, and for example, tablets (including orally disintegrating tablets, chewable tablets, troche tablets, etc.), granules, powders, soft capsules, hard capsules, troches, and jellies. Oral agents such as agents or liquids (including suspensions, emulsions, syrups, etc.); external agents such as ointments, suppositories, patches, sprays; injections, etc. may be mentioned. Among these, from the viewpoint of ease of handling and further enhancing the effect of the present invention, oral preparations are preferable, and tablets, granules, soft capsules, and hard capsules are more preferable.

The agent according to this embodiment can be used as a component of, for example, pharmaceuticals, quasi-drugs, cosmetics, and foods and drinks (beverages, foods). In addition, the agents according to the present embodiment include, for example, pharmaceutical preparations, non-pharmaceutical preparations, foods for specified health use, foods with nutritional functions, foods for the elderly, foods for special purposes, foods with functional claims, health supplements (supplements), etc. It can also be used as a dietary supplement (eg, confectionery tablets), apparently a food. Furthermore, the agent according to this embodiment can also be used as, for example, a veterinary drug or a feed additive.

When the agent according to this embodiment is used as a food, the food may be a general food containing the component (A) and, if necessary, other components. Examples of such foods include solid foods such as cookies, biscuits, snacks, jellies, gummies, chocolates, gums, candy and cheese; and liquid foods such as energy drinks, juices, tea beverages, coffee beverages and milk beverages.

The agent according to this embodiment is a subject (for example, human; cow, pig, horse, sheep, goat, dog, rabbit, mouse, rat, guinea pig, etc.) that requires an action of suppressing the progress of aging of female reproductive tissues. It can be suitably used for non-human animals such as gerbils, hamsters and ferrets). As a target, a human is preferable from the viewpoint of further enhancing the effect of the present invention. Further, from the viewpoint of further enhancing the effect of the present invention, cows, pigs and horses are preferable among non-human animals, and pigs are more preferable.

[2. How to control the aging process of ovaries, follicles or eggs]
Pyrroloquinoline quinone or a salt thereof has an action of suppressing the progress of aging of ovaries, follicles or ova. Therefore, as one embodiment of the present invention, there is provided a method of suppressing the progress of aging of an ovary, a follicle or an egg by ingesting a composition containing pyrroloquinoline quinone or a salt thereof. The ovary, follicle or egg may be a human ovary, follicle or egg and is a non-human animal such as cow, pig, horse, sheep, goat, dog, rabbit, mouse, rat, guinea pig, snail, hamster, ferret. It may be an ovary, a follicle or an egg. From the viewpoint of further enhancing the effect of the present invention, it is preferably a human ovary, follicle or egg. Further, from the viewpoint of further enhancing the effect of the present invention, among the ovaries, follicles or eggs of non-human animals, bovine, pig or horse ovaries, follicles or eggs are preferable, and pig ovaries, follicles or eggs are more preferable.

In addition, in these embodiments, the type, content, daily intake, etc. of pyrroloquinoline quinone or a salt thereof in the composition, the type, content, daily intake, etc. of other components, etc. For the formulation form of the product, see [1. Anti-aging agents for ovaries, follicles or eggs].

Hereinafter, the present invention will be described in more detail based on Examples and the like. 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.

[Test Example 1: Effect of administration of pyrroloquinoline quinone on blood AMH (anti-Müllerian hormone) in female mice]
Six-month-old Leydig cell-specific NRG1-deficient female mice were treated with PQQ disodium salt for 2 weeks (2 mg / kg / day) (n = 5). In addition, 6-month-old Leydig cell-specific NRG1-deficient male mice (KO; control) and 6-month-old wild-type mice (WT mice) were each administered with water alone for 2 weeks. Then, blood was collected and the concentration of AMH (anti-Müllerian hormone), which is a follicle marker, was measured using an AMH ELISA kit (Elabscience Biotechnology, Bethesda, MD) according to the method of use of the kit. The results are shown in FIG. The Leydig cell-specific NRG1-deficient female mouse has a short life span, and 6 months of age corresponds to the human 40s.

As shown in FIG. 1, control female mice not administered PQQ disodium salt had a lower concentration of AMH than WT mice, whereas female mice administered PQQ disodium salt had AMH compared to WT mice. Concentration increased. That is, it was confirmed that the ovarian tissue was rejuvenated by ingestion of PQQ disodium salt.

[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) continued to receive PQQ disodium salt in drinking water (2 mg / kg / day). Female mice to which only water was administered were used as controls. The estrous cycle of each mouse was examined by vaginal dirt examination. The vaginal dirt test was performed as follows: The inside of the vagina was refluxed with physiological saline, and the refluxed substance was smeared on a slide glass. After drying, it was fixed with methanol and stained with Giemsa. This was observed under an inverted microscope (manufactured by Keyens), and when only leukocytes were observed, the estrus resting period was observed, when only the epithelial cells with nuclei were observed, the preestrus, and only the anucleated epithelial cells were observed. The case is in estrus, a mixture of anucleated epithelial cells and leukocytes, and the number of annuclear epithelial cells is higher in the microestrus, a mixture of annuclear epithelial cells and leukocytes, and the number of leukocytes When more people were seen, it was judged to be late estrus. The results are shown in FIG.

As shown in FIG. 2, control female mice not administered PQQ disodium salt always maintained a microestrus period, and no periodic change in estrus was observed, whereas PQQ disodium salt was administered. In female mice, it was confirmed that the cyclical change in estrus resumed about 14 days after the start of administration. That is, it was confirmed that the administration of PQQ disodium salt improved the abnormal ovulation cycle of female mice.

[Test Example 3: Effect of administration of pyrroloquinoline quinone on eggs]
Three-week-old immature female mice were given PQQ disodium salt in drinking water (2 mg / kg / day) (n = 5). Female mice to which only water was administered were used as controls. Two days after the start of administration, eCG (pregnant horse serum gonadotropin) 4IU, which induces follicle development, was administered, and 48 hours later, hCG (human chorionic gonadotropin) 5IU was administered. It was administered. Then, the number of ovulations of each female mouse was measured. The number of ovulations is determined by collecting the fallopian tubes from the slaughtered mice, incising the ampulla of the fallopian tubes, collecting the ovulated egg-cumulus cell complex, and observing the complex visually under a microscope. Calculated. In vitro fertilization was performed using the ovulated ovum, and the fertilization rate and the incidence rate were calculated. Here, the fertilized egg refers to a split egg 24 hours after fertilization, and the fertilization rate refers to the ratio of the number of successfully fertilized eggs to the total number of mature eggs ovulated. The incidence refers to the proportion of fertilized eggs that have become blastocyst stage embryos. Table 1 shows the results of the number of ovulations and the fertilization rate expressed by the mean ± standard deviation. In addition, 3 weeks of age in mice corresponds to the teens when menstruation begins in humans.

As shown in Table 1, female mice administered with PQQ disodium salt maintained in vitro fertilization rates while increasing ovulation numbers and maintained in vitro fertilization rates as compared to control female mice not administered PQQ disodium salt. The incidence improved while maintaining the in vitro fertilization rate. That is, it was confirmed that the quality of the egg was improved by the administration of PQQ disodium salt.

[Test Example 4: Effect of administration of pyrroloquinoline quinone on blood female hormone concentration]
Three-week-old immature female mice were given PQQ disodium salt in drinking water (2 mg / kg / day). Female mice to which only water was administered were used as controls. Two days after the start of administration, eCG (pregnant horse serum gonadotropin) 4IU, which induces follicle development, was intraperitoneally administered, and 48 hours later, hCG (human chorionic gonadotropin) 5IU. Was intraperitoneally administered. Serum of each female mouse was collected over time immediately before administration of eCG, and blood estradiol (E2) concentration as a follicle development marker and progesterone (P4) concentration as a luteal marker were measured in Endocrine Technology, Inc. Measurements were made using a commercially available kit from (Newark, CA). The results are shown in FIG.

As shown in FIG. 3 (A), the blood estradiol concentration was higher in the female mice to which the PQQ disodium salt was administered than in the control female mice to which the PQQ disodium salt was not administered. That is, it was confirmed that administration of PQQ disodium salt promoted the production of estradiol. Estradiol is one of the female hormones widely used in hormone replacement therapy, and it is widely known that it is used for the treatment of menopausal disorders and the improvement of menopausal symptoms. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof is useful as an ameliorating agent or a preventive agent for symptoms derived from estrogen deficiency in females, and as an ameliorating agent or a preventive agent for pre-menopausal disorders or climacteric disorders in females. This allows pre-menopausal or menopausal women to reach menopause without pain.
In addition, as shown in FIG. 3 (B), no significant difference was observed in the increase in blood progesterone concentration between the female mouse to which PQQ disodium salt was administered and the control female mouse to which PQQ disodium salt was not administered. .. Progesterone is known to be involved in blood enrichment in ovarian hyperstimulation syndrome. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof is useful as a preventive agent for ovarian hyperstimulation syndrome by relaxing the production of progesterone while promoting the production of estradiol.

[Test Example 5: Effect of administration of pyrroloquinoline quinone on ovarian fibrosis]
Six-month-old Leydig cell-specific NRG1-deficient female mice (KO) were treated with PQQ disodium salt for 2 weeks (2 mg / kg / day). Then, the removed ovary was stained with PSR and observed with an inverted microscope (manufactured by KEYENCE) at a magnification of 20 times. The results are shown in FIG.

As shown in FIG. 4, in the mice before PQQ disodium salt administration, the inside of the stromal cells was dyed red as a whole, so that collagen deposition, that is, ovarian fibrosis was confirmed. At the same time, atrophy of interstitial follicles was also observed. On the other hand, in the mice to which PQQ disodium salt was administered for 2 weeks, it was confirmed that the red color of the stromal cells disappeared and the fibrotic stromal cells were improved to a healthy state. It is known that stromal cells of the ovary become fibrotic with aging. That is, it was confirmed that the ovarian tissue was rejuvenated by ingestion of PQQ disodium salt. In addition, antral follicles were observed, and it was confirmed that the atrophy of the follicles was also improved, confirming that the quality of the ova was improved.

From the results of Test Examples 1 to 5, by ingesting pyrroloquinoline quinone or a salt thereof, it is possible to maintain the QOL of women who tend to be impaired due to mental and physical abnormalities that occur as menopause approaches, and women can be vibrant and young even when they are old. It can be said that you can spend your time well.

Claims

An ovarian, follicle or egg anti-aging agent containing pyrroloquinoline quinone or a salt thereof.
An inhibitor or ameliorating agent for fibrosis of female reproductive tissues, which contains pyrroloquinoline quinone or a salt thereof.
An egg quality improver containing pyrroloquinoline quinone or a salt thereof.
An improving or preventing agent for menstrual irregularities containing pyrroloquinoline quinone or a salt thereof.
An improving agent or a preventive agent for ovarian hyperstimulation syndrome containing pyrroloquinoline quinone or a salt thereof.
An estrogen production promoter containing pyrroloquinoline quinone or a salt thereof.
The agent according to claim 6, which improves or prevents symptoms caused by female estrogen deficiency.
The agent according to claim 6, which improves or prevents pre-menopausal disorders or menopausal disorders in females.
The agent according to claim 6, which improves or prevents menopausal symptoms in females.
The agent according to any one of claims 1 to 9, for use in non-human animals.
A method of suppressing the progress of aging of ovaries, follicles or eggs by ingesting a composition containing pyrroloquinoline quinone or a salt thereof.