CN114007618A

CN114007618A - Female anti-aging agent

Info

Publication number: CN114007618A
Application number: CN202080045230.3A
Authority: CN
Inventors: 岛田昌之
Original assignee: Hiroshima University NUC; Rohto Pharmaceutical Co Ltd
Current assignee: Hiroshima University NUC; Rohto Pharmaceutical Co Ltd
Priority date: 2019-06-28
Filing date: 2020-06-22
Publication date: 2022-02-01
Also published as: TW202114674A; JPWO2020262325A1; WO2020262325A1

Abstract

The present invention relates to an anti-aging agent for ovary, follicle or ovum, which contains pyrroloquinoline quinone or a salt thereof.

Description

Female anti-aging agent

Technical Field

The invention relates to a female anti-aging agent.

Background

In women, reproductive tissues such as ovaries, follicles, ova, and the like gradually age with age, and become difficult to be pregnant or delivered. For the treatment of infertility in such women with difficulty in pregnancy, hormone replacement therapy is mostly used to promote ovulation. However, in addition to the difficulty in developing and discharging uniformly mature follicles, a symptom called ovarian hyperstimulation syndrome (OHSS) may appear as a side effect, and if the symptom worsens, there is a risk of falling into respiratory failure or circulatory failure. Therefore, a safe and effective treatment method for infertility treatment is being sought (non-patent document 1).

In addition, as female reproductive tissues undergo aging, if the concentration of female hormones such as estrogen is decreased, menopausal disorders are mainly induced in women aged 40 to 50 years old. Among them, in women aged 30 to 40, in addition to the symptoms of the above-mentioned climacteric disorder, irregular menstruation occurs in addition to stress or fatigue. This is called a pre-menopausal disorder, which, like climacteric disorders, significantly reduces the QOL (quality of life) of women. In the treatment of these menopausal disorders or pre-menopausal disorders, hormone replacement therapy is mainly performed, but there is a problem that there is a possibility that cancer is caused or side effects such as irregular genital bleeding, vomiting, headache, liver dysfunction, and the like are caused. For any woman, there is a possibility of causing an premenstrual or climacteric disorder, and especially in japan where it is advanced to age, it is very important that women transition from premenstrual to menopause in a healthy manner. Therefore, as in the case of infertility treatment, a safe and effective treatment method for premenstrual and climacteric disorders is being sought (non-patent document 2).

However, aging of female reproductive tissues is becoming a problem not only in humans but also in livestock such as cattle and swine. For example, in the pig industry, sows pass from mating to farrowing 2.5 times per year, but with age, the litter size of sows decreases, and reproductive performance deteriorates due to increased reproductive failure such as abortion. Therefore, it is desirable to develop a method for inhibiting the aging process of female reproductive tissues in non-human animals in addition to humans.

Documents of the prior art

Non-patent document

Non-patent document 1: japanese coal , 1998, volume 50, No. 6, N135-N138

Non-patent document 2: JIM, 1999, Vol.9, No. 8, 688-

Disclosure of Invention

Technical problem to be solved by the invention

The present invention aims to provide an anti-aging agent for an ovary, follicle, or ovum (hereinafter sometimes collectively referred to as "female reproductive tissue").

Means for solving the problems

The inventors of the present application found that pyrroloquinoline quinone or a salt thereof can inhibit the aging process of female reproductive tissues and can promote the production of estradiol. The present invention has been completed based on this new finding.

The present invention provides, for example, the following inventions.

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

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

[3] An egg quality improving agent comprising pyrroloquinoline quinone or a salt thereof.

[4] An ameliorating or prophylactic agent for menoxenia, which comprises pyrroloquinoline quinone or a salt thereof.

[5] An ameliorating or prophylactic agent for ovarian hyperstimulation syndrome, which comprises pyrroloquinoline quinone or a salt thereof.

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

[7] The estrogen production promoter according to [6], which improves or prevents symptoms caused by estrogen deficiency in females.

[8] The estrogen production promoter according to [6], which improves or prevents a female's pre-menopausal or menopausal disorder.

[9] The estrogen production promoter according to [6], which improves or prevents female climacteric symptoms.

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

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

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

[2-2] the anti-aging agent according to [1], which improves ovum quality.

[2-3] the anti-aging agent according to [1], which improves or prevents irregular menstruation.

[2-4] the anti-aging agent according to [1], which improves or prevents ovarian hyperstimulation syndrome.

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

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

[2-7] the antiaging agent according to [2-5], which improves or prevents a female's pre-menopausal or menopausal disorder.

[2-8] the antiaging agent according to [2-5], which improves or prevents female climacteric symptoms.

Effects of the invention

The present invention can provide an anti-aging agent for female reproductive tissues.

Drawings

Fig. 1 is a graph showing the blood AMH concentration of each female mouse in test example 1.

Fig. 2 (a) is a graph showing the estrous cycle when pyrroloquinoline quinone disodium salt was not administered to aging model mice for females in test example 2. Fig. 2 (B) is a graph showing the estrous cycle when pyrroloquinoline quinone is administered to an aging model mouse for females in test example 2. In fig. 2, M denotes the postestrus, WE denotes the micro estrus, E denotes the estrus, P denotes the pre-estrus, and D denotes the estrus.

Fig. 3 (a) is a graph showing the relationship between the time and the blood estradiol concentration in experimental example 4 when eCG (equine chorionic gonadotropin) is administered to an immature female mouse to which pyrroloquinoline quinone disodium salt is administered or not administered. Fig. 3 (B) is a graph showing the relationship between the time and the blood xanthone concentration in the immature female mice administered with or without administration of pyrroloquinoline quinone disodium salt in test example 4 and the administration of eCG (equine chorionic gonadotropin) and hCG (human chorionic gonadotropin).

Fig. 4 (a) is a photograph (magnification 20 times) of an ovary of an aging model mouse female before administering pyrroloquinoline quinone disodium salt in test example 5 after PSR staining. Fig. 4 (B) is a photograph (magnification: 20 times) of an ovary stained with PSR after 2 weeks of administration of pyrroloquinoline quinone disodium salt to a female aging model mouse in test example 5. Fig. 4 (C) and 4 (D) are diagrams obtained by tracing (tracing) fig. 4 (a) and 4 (B), respectively, and the hatched regions indicate the dyed regions.

Detailed Description

Hereinafter, specific embodiments of the present invention will be described in detail. However, the present invention is not limited to the following embodiments.

[1. anti-aging agent for ovary, follicle or ovum ]

The ovarian, ovarian follicle, or ovum anti-aging agent of the present embodiment contains pyrroloquinoline quinone or a salt thereof (also referred to as "(a) component").

Pyrroloquinoline quinone or a salt thereof exerts an effect of suppressing disappearance or decline of AMH (Anti-Mullerian Hormone) in females. Here, AMH is a hormone that serves as 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 exerts an effect of inhibiting or improving fibrosis of female reproductive tissues (ovary, follicle, ovum, etc., particularly ovary). Accordingly, as an embodiment of the present invention, there is provided an anti-aging agent for ovary, follicle or ovum, and an inhibitor or improver for fibrosis of female reproductive tissue, which contain pyrroloquinoline quinone or a salt thereof. The term "anti-aging" as used herein means that a woman recovers youth, suppresses the aging process of a woman, maintains high QOL even in an elderly woman, and is active and youth even in an elderly woman. The term "advanced age" as used herein means an age of 30 or more, preferably 40 or more, and more preferably 50 or more. In addition, as an example of senescence of female reproductive tissues (particularly, ovarian tissues), ovarian reserve function (the number of remaining secondary follicles in the ovary) can be cited. As described above, pyrroloquinoline quinone or a salt thereof has an effect of inhibiting disappearance or decrease of AMH in females, and thus has an effect of improving ovarian reserve function.

Pyrroloquinoline quinone or a salt thereof exerts an effect of improving the quality of an ovum. Accordingly, in one embodiment of the present invention, an ovum quality improving agent containing pyrroloquinoline quinone or a salt thereof is provided. The term "improvement in the quality of an egg" as used herein means increasing the number of ovulations and maintaining or improving the fertilization rate of the discharged egg, and the fertilization rate is a ratio of the number of fertilized eggs to the total number of discharged eggs. Accordingly, as one embodiment of the present invention, there is provided a preparation containing pyrroloquinoline quinone or a salt thereof for increasing the number of ova to be discharged and maintaining or improving the fertilization rate of the discharged ova. The term "improvement in the quality of an ovum" means maintaining or improving the fertilization rate of an ovum and maintaining or improving the development rate, and the development rate as used herein means the proportion of a fertilized ovum that is a blastocyst in the fertilized ovum. Accordingly, as one embodiment of the present invention, there is provided an agent for maintaining or increasing the fertilization rate and the development rate of an ovum, which comprises pyrroloquinoline quinone or a salt thereof.

Pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing irregular menstruation. Accordingly, an embodiment of the present invention provides an ameliorating or prophylactic agent for menstrual disorder, which comprises pyrroloquinoline quinone or a salt thereof. Here, "irregular menstruation" refers to a state in which the menstrual cycle of a female (ovulation if a female non-human animal is amenorrhea) is abnormal. For example, in humans, irregular menstruation is considered to be a period of normal menstruation of 25 to 38 days, a period of no menstruation of 24 days or less, or 39 days or more, or 3 months or more.

Pyrroloquinoline quinone or a salt thereof exerts an effect of promoting the production of estrogen (e.g., estrone, estradiol, estriol, and preferably estradiol). Accordingly, as one embodiment of the present invention, there is provided an estrogen production promoter containing pyrroloquinoline quinone or a salt thereof. Pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing female symptoms caused by estrogen deficiency, an effect of improving or preventing female climacteric symptoms, an effect of improving or preventing female premenstrual climacteric disorder, or an effect of improving or preventing female climacteric disorder by promoting estrogen production, and in addition, exerts an effect of supplementing female hormones. Accordingly, an embodiment of the present invention provides an ameliorating or prophylactic agent for female symptoms caused by estrogen deficiency, an ameliorating or prophylactic agent for female climacteric symptoms, or an ameliorating or prophylactic agent for female premenstrual tension disorders or climacteric disorders, which contains pyrroloquinoline quinone or a salt thereof.

Pyrroloquinoline quinone or a salt thereof exerts an effect of improving or preventing ovarian hyperstimulation syndrome. Accordingly, as an embodiment of the present invention, there is provided an ameliorating or prophylactic agent for ovarian hyperstimulation syndrome, which comprises pyrroloquinoline quinone or a salt thereof. Here, the "ovarian hyperstimulation syndrome" refers to various symptoms caused by swelling of ovaries due to hyperstimulation by an ovulation promoting agent, and then, abdominal dropsy or pleural effusion, and in the case of severe cases, blood concentration may occur, which may cause complications such as renal failure and thrombosis. Specific symptoms of ovarian hyperstimulation syndrome include abdominal distension, nausea, rapid weight gain, decreased urine volume, abdominal pain, and diarrhea. In addition, pyrroloquinoline quinone or a salt thereof exerts an effect of ameliorating or preventing symptoms caused by ovarian hyperstimulation syndrome and complications of ovarian hyperstimulation syndrome by ameliorating or preventing ovarian hyperstimulation syndrome. The preparations containing pyrroloquinoline quinone or a salt thereof according to the above embodiments are collectively referred to as "preparations according to the present embodiment".

In the present specification, "menopause" means that a female is in menopause within about 10 years before and after menopause, and in the case of humans, during the later period of 40 years to 50 years, the period is in menopause. In the present specification, the term "premenopausal period" refers to a period younger than the menopause and at which symptoms of the aforementioned premenopausal disorder appear, and in humans, a period in the middle of 30 to 40 years of age belongs to the premenopausal period.

Examples of the symptoms of the "female climacteric disorder" include dyspnea, dizziness, vertigo, quadriplegia, stiffness in limbs, edema, fatigue, tinnitus, head congestion, hot flashes (hot flashes), sweating, palpitation, excitement, insomnia, anxiety, irritability, emotional instability, feelings of weakness, easy tearing, low heat, anxiety, headache, shoulder soreness, chest pain, lumbago, arthralgia, cold limbs, itching, dryness of skin or eyes, nausea, anorexia, abdominal pain, constipation, diarrhea, frequent urination, sexual intercourse disorder, and vulvar discomfort.

Examples of the "female symptoms caused by estrogen deficiency" include menstrual disorder, autonomic imbalance, psychonervous system symptoms, urogenital atrophy, hyperlipidemia, arteriosclerosis, hypertension, stroke, osteoporosis, osteopenia, skin atrophy, arthralgia, dry mouth, taste change, sleep disorder, and symptoms of the "female climacteric disorder" described above, and in addition, symptoms described in "expected action and effect of 1. HRT" in "hormone supplement therapy guideline 2017 edition" edited by the japan society of obstetrics and gynecology.

The symptoms of "premenstrual climacteric disorder (female youth-related climacteric disorder)" include, in addition to the symptoms of the above-mentioned "female climacteric disorder", irregular menstruation, amenorrhea, and the like.

The "female climacteric symptom" refers to the symptom of the "female climacteric disorder" or the symptom of the "premenstrual climacteric disorder (female juvenile climacteric disorder)". However, not only "menopause" or "premenopause," this condition may also occur in young years (e.g., the teens years of human menstruation). The "female climacteric symptom" may be a slight symptom among the above-mentioned specific symptoms. Examples thereof include a state in which the number of hot flashes or sweating is small although the hot flashes or sweating is sensed by itself, and a state in which depression or shoulder soreness is small.

Pyrroloquinoline quinone is a known compound represented by the following formula.

[ chemical formula 1]

Examples of the salt of pyrroloquinoline quinone 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 salts and magnesium salts.

The pyrroloquinoline quinone or a salt thereof is preferably an alkali metal salt of pyrroloquinoline quinone, more preferably a sodium salt of pyrroloquinoline quinone, and still more preferably a disodium salt of pyrroloquinoline quinone.

Commercially available products can be used for pyrroloquinoline quinone or a salt thereof. In addition, as the pyrroloquinoline quinone or a salt thereof, for example, a crop containing pyrroloquinoline quinone or a salt thereof, such as natto, soybean, cocoa powder, cacao mass, cacao, parsley, or green pepper, can be used.

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

The formulation of the present embodiment may vary in the daily dose of component (a) depending on the condition (body weight, age, sex, etc.) of the individual to be ingested, the form of the formulation, etc., and when used in humans, the daily dose of component (a) is preferably 7 to 100mg, more preferably 10 to 40mg, even more preferably 20 to 25mg, and when used in non-human animals, it is preferably 1 to 30mg/kg, even more preferably 2 to 20mg/kg, from the viewpoint of ease of ingestion in a single dose, from the viewpoint of effectiveness based on the physiological action of component (a), and from the viewpoint of safety.

The preparation of the present embodiment may further contain a polyamine (also referred to as "component (B)") and a crude drug (also referred to as "component (C)"). This can bring about more remarkable effects of the present invention.

Polyamines are aliphatic hydrocarbons containing more than 2 primary amino groups in the molecule. Specific examples of the polyamine include putrescine, cadaverine, ethylenediamine, trimethylenediamine, hexamethylenediamine, spermidine (caldine), homopiperazine (homosperamine), aminopropylcadaverine, spermine, thermomylamine (thermamine), thermoptamine (thermospirmine), canavalimine (canavaline), aminopentylgramine (aminopentyl norspermine), aminopropyl homopiperazine (aminopropyl homospermine), canavaline, homopiperazine, thermophilic pentamine (caltropamine), thermophilic pentamine (homocaltropamine), thermophilic pentamine (homocaltropamide), aminopropyl canavalinamine, bis (aminopropyl) homoprimine, bis (aminopropyl) norspermine (bis (amidopropypro) homoprimine), aminobutyltetramine, aminopropyl homoprimine, homopentamine (homopentavalienamine), homopentaxanthamine (homopentaxanthamine), thermophilic hexamine (thermophilic hexamine), thermophilic hexamine (thermoamine). From the viewpoint of ease of use, spermidine, spermine, and putrescine are preferable as the polyamine, spermidine and spermine are more preferable, and spermidine is further preferable.

Commercially available products can also be used as the polyamine. One of the polyamines may be used alone, or two or more of them may be used in combination. Further, as the polyamine, a polyamine-containing composition (also referred to as "polyamine composition") such as an extract from a plant such as soybean (preferably soybean germ), wheat (preferably wheat germ), or rice (preferably rice germ), an extract from fish or shellfish (preferably milt), or dry sake yeast can be used. When the polyamine composition is an extract derived from a plant, water, an organic solvent such as ethanol, or a mixed solvent of these solvents can be used as the extraction solvent. From the viewpoint of ease of use, as the polyamine composition, an extract derived from a plant is preferable, and an extract derived from soybean (preferably soybean germ) is more preferable. Examples of commercially available POLYAMINE compositions include "Soypolyya" (manufactured by Combi Corporation), "ORYZA POLYAMINE-P" (manufactured by Oryza Oil & Fat Chemical Co., manufactured by Ltd), "ORYZA POLYOLYAMINE-LC" (manufactured by Oryza Oil & Fat Chemical Co., manufactured by Ltd), "PHYTOPOLYAMINE-S" (manufactured by TOYOBO CO., manufactured by LTD.), "PHYTOPOLYAMINE-SP" (manufactured by TOYOBO CO., manufactured by LTD.), "OYOPOLYAMINE-SP" (manufactured by MITSUBISHI GAS CHEMICAL COMPANY, manufactured by INC.), and the like.

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

When the polyamine composition is contained as the component (B) in the preparation of the present embodiment, the content of the polyamine composition is not particularly limited, and may be appropriately set according to the type of the polyamine composition, the type and content of other blending components, the preparation form, and the like. From the viewpoint of stability of the preparation, the viewpoint of effectiveness based on physiological action of the component (B), safety, and the viewpoint of reducing the influence of the odor contained in the component (B), the total content of the polyamine composition is preferably 5 to 35% by weight, more preferably 10 to 30% by weight, and further preferably 14 to 23% by weight, based on the total amount of the preparation of the present embodiment, for example, as the content of the polyamine composition.

The content ratio of the component (B) to the component (a) in the preparation of the present embodiment is not particularly limited, and may be appropriately set depending on the types of the component (a) and the component (B), the types and contents of other blending components, the preparation form, and the like. From the viewpoint of further improving the effect of the present invention, the content ratio of the component (B) to the component (a) is, for example, preferably 0.001 to 0.1 part by weight, more preferably 0.01 to 0.03 part by weight, and further preferably 0.015 to 0.02 part by weight, based on 1 part by weight of the total content of the component (a) contained in the preparation of the present embodiment.

In the preparation of the present embodiment, when the polyamine composition is contained as the component (B), the content ratio of the polyamine composition to the component (a) is not particularly limited, and may be appropriately set depending on the types of the component (a) and the polyamine composition, the types and contents of other blending components, the preparation form, and the like. From the viewpoint of further enhancing the effect of the present invention, the content ratio of the polyamine composition to the component (a) is, for example, preferably 5 to 15 parts by weight, more preferably 7 to 10 parts by weight, based on 1 part by weight of the total content of the component (a) contained in the preparation of the present embodiment.

In the preparation of the present embodiment, when the component (B) contains spermidine and spermine, the weight ratio of spermine to spermidine is not particularly limited, and can be appropriately set according to the types of the components (a) and (B), the types and contents of other blending components, the preparation form, and the like. From the viewpoint of further improving the effect 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.

In the preparation of the present embodiment, the daily intake amount of component (B) differs depending on the state (body weight, age, sex, etc.) of the subject to be taken, the form of the preparation, etc., but is preferably 0.001 to 20mg, more preferably 0.01 to 2mg, further preferably 0.1 to 0.7mg, particularly preferably 0.3 to 0.4mg, from the viewpoints of effectiveness based on the physiological action of component (B), safety, and reduction of the influence of the odor of component (B).

In the preparation of the present embodiment, when the polyamine composition is contained as the component (B), the daily intake amount of the polyamine composition varies depending on the state (body weight, age, sex, etc.) of the subject to be ingested, the form of the preparation, etc., but is preferably 50 to 400mg, more preferably 100 to 300mg, further preferably 200 to 250mg, particularly preferably 201 to 220mg, from the viewpoint of ease of single dose ingestion, the viewpoint of effectiveness based on the physiological action of the component (B), safety, and the viewpoint of reducing the influence of the odor of the component (B).

The crude drug is a natural product such as a plant, an animal, or a mineral having a medicinal purpose, or a natural product such as a plant, an animal, or a mineral, which is obtained by simply processing the whole or a part of the natural product such as drying. In the preparation of the present embodiment, examples of the form of crude drugs include birth drugs (crude drugs); drying and pulverizing crude drug to obtain crude drug powder; crude drug extract obtained by extracting crude drug or crude drug powder with water, or organic solvent such as ethanol, or mixed solvent of these solvents. Among them, crude drug extracts are preferred from the viewpoint of more remarkably exerting the effects of the present invention. The crude drug extract may be the extract itself (tincture, fluid extracts, etc.), a product obtained by diluting or concentrating the extract (viscous extracts, etc.), or a product obtained by drying the extract and pulverizing the extract or making the extract into paste (dry extracts, etc.).

Specific examples of the crude drug extract include, for example, a ginger extract, a maca extract, a licorice extract, a astragalus extract, a tangerine peel extract, a peony extract, a ginseng extract, a turmeric extract, a cistanche extract, a cinnamon extract, an acanthopanax extract, a hawthorn extract, a rehmannia extract, an angelica extract, a saw palmetto extract, a ginkgo leaf extract, a bupleurum extract, a cimicifuga foetida extract, a yam extract, a jujube extract, a atractylodes macrocephala extract, a atractylodes lancea extract, a ganoderma extract, a deer antler extract, a eurycoma longifolia extract, a french maritime pine bark extract, a quercus robur extract, a dioscorea villosa extract, a dioscorea opposita extract, a cistanche tubulosa extract, a garlic extract, an acanthopanax extract, a velvet bean extract, a gardenia extract, a deli extract, a cordyceps sinensis mycelium powder, a ginseng extract, a bark extract, a ginseng extract, a ginseng extract, a ginseng extract, a ginseng extract, a, Black ginger extract, black pepper extract, pomegranate seed extract, vitex extract, black cohosh extract, dandelion extract, etc. From the viewpoint of stability of the preparation, preferred crude drug extracts are maca extract, ginseng extract, serenoa repens extract, ginger extract, eurycoma longifolia extract, french maritime pine bark extract, quercus robur extract, dioscorea villosa extract, dioscin-containing sweet potato extract, pomegranate seed extract, and vitex negundo extract, and more preferred maca extract and ginseng extract.

The crude drug may be a commercially available product. The crude drugs can be used singly or in combination of two or more.

The content of the component (C) in the preparation of the present embodiment is not particularly limited, and may be appropriately set depending on the kind of the component (C), the kinds and contents of other blending components, the preparation form, and the like. From the viewpoint of further remarkably exhibiting the effects of the present invention, the total content of the component (C) is preferably 0.1 to 70% by weight, more preferably 1 to 10% by weight, and still more preferably 2 to 4% by weight, based on the total amount of the preparation of the present embodiment, for example.

The content ratio of the component (C) to the component (a) in the preparation of the present embodiment is not particularly limited, and may be appropriately set depending on the types of the component (a) and the component (C), the types and contents of other blending components, the preparation form, and the like. From the viewpoint of further improving the effect of the present invention, the content ratio of the component (C) to the component (a) is, for example, 1 part by weight of the total content of the component (a) contained in the preparation of the present embodiment, and 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 still more preferably 1 to 1.5 parts by weight.

The content ratio of the component (C) to the component (B) in the preparation of the present embodiment is not particularly limited, and may be appropriately set depending on the types of the component (B) and the component (C), the types and contents of other blending components, the preparation form, and the like. From the viewpoint of further improving the effect of the present invention, the content ratio of the component (C) to the component (B) is, for example, 1 part by weight of the total content of the component (B) contained in the preparation of the present embodiment, and the total content of the component (C) is preferably 50 to 250 parts by weight, more preferably 80 to 200 parts by weight, and further preferably 100 to 170 parts by weight.

The preparation of the present embodiment may contain pharmacologically active ingredients or physiologically active ingredients other than the component (a), the component (B), and the component (C) within a range not to impair the effects of the present invention. Specific examples of the pharmacologically active ingredient or physiologically active ingredient include ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, folic acid, vitamin B12, vitamin C, vitamin D, vitamin a, vitamin E, nicotinic acid, calcium pantothenate, taurine, carnosine, anserine, whale carnosine, citrulline, gamma-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, carnitine, glutamic acid, glutamine, 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, extract of placenta, placenta hominis, placenta, and the like, Pycnogenol (procyanidins), equol, etc. From the viewpoint of further improving the effect of the present invention, as the pharmacologically active ingredient or physiologically active ingredient, ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, resveratrol, isoflavone, equol, pycnogenol (procyanidin), folic acid are preferable, and ubiquinone (coenzyme Q10), astaxanthin, zinc, resveratrol, isoflavone, equol, pycnogenol (procyanidin), folic acid are more preferable. The pharmacologically active ingredient or the physiologically active ingredient may be used alone or in combination of two or more.

The formulation of the present embodiment may contain various additives other than the above-described components within a range not to impair the effects of the present invention. Specific examples of the additives include excipients, binders, disintegrating agents, lubricants, coloring agents, flavoring agents, deodorizing agents, saccharides, sugar alcohols/polyols, high-intensity sweeteners, fats and oils, emulsifiers, thickeners, souring agents, fruit juices, and the like. Examples of the excipient include lactose, white sugar, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, silicic acid, and the like. Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, calcium phosphate, and polyvinylpyrrolidone. Examples of the disintegrating agent include dried starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, glycerol monostearate, and lactose. Examples of the lubricant include talc, stearate, borax, and polyethylene glycol. Examples of the taste-improving agent include white sugar, orange peel, citric acid, tartaric acid, and the like. Examples of the saccharide include sugars such as sucrose, isomerized sugar, glucose, fructose, palatinose, trehalose, lactose, and xylose. Examples of the sugar alcohol/polyol include sorbitol, xylitol, erythritol, lactitol, isomalt, hydrogenated syrup, hydrogenated maltose syrup, glycerol, and propylene glycol. Examples of the high-intensity sweetener include aspartame, stevia, acesulfame potassium, and sucralose. Examples of the oils and fats include safflower oil (safflower oil), grape seed oil, sunflower seed oil (sunflower oil), olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, and perilla oil. Examples of the emulsifier include sucrose fatty acid esters, glycerin fatty acid esters, and lecithin. Examples of the thickener include carrageenan, xanthan gum, guar gum, pectin, and locust bean gum. Examples of the acidulant include citric acid, lactic acid, and malic acid. Examples of the fruit juice include lemon juice, orange juice, and berry juice. One kind of the additive may be used alone, or two or more kinds may be used in combination. From the viewpoint of stability of the components (a) and (B) during formulation, it is preferable that the additive contains an oil or fat.

The dosage form of the preparation of the present embodiment is not particularly limited, and examples thereof include oral preparations such as tablets (including orally disintegrating tablets, chewable tablets, buccal tablets, and the like), granules, powders, soft capsules, hard capsules, buccal tablets, jellies (jellies), and liquid preparations (including suspensions, emulsions, syrups and the like); external preparations such as ointment, suppository, patch, and spray; injections, and the like. Among them, from the viewpoint of easy handling and further improvement of the effect of the present invention, oral preparations are preferable, and tablets, granules, soft capsules and hard capsules are more preferable.

The preparation of the present embodiment can be used, for example, as a component of a drug, a quasi drug (quasi drug), a cosmetic, a food or drink (beverage, food). Further, the agent of the present embodiment is useful, for example, as a pharmaceutical preparation, a quasi drug preparation, a specific health food, a nutritional functional food, an elderly food, a special-use food, a functional food, a health supplement (a nutritional supplement), a food preparation (e.g., a candy tablet), a self-evident food (a ming らか food). Further, the preparation of the present embodiment can be used, for example, as a drug for animals or a feed additive.

When the preparation of the present embodiment is used as a food, the food may be a food obtained by blending the component (a) and, if necessary, other components with a usual food. Examples of such food products include solid food products such as cookies, crackers, snacks, jellies, soft sweets, chocolates, chewing gums, cerealose, and cheese; liquid food such as nutritious beverage, fruit juice, tea beverage, coffee beverage, and milk beverage.

The formulation of the present embodiment can be suitably used for subjects (e.g., humans; non-human animals such as cows, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, ferrets, etc.) in need of an effect of inhibiting the aging process of female reproductive tissues. From the viewpoint of further improving the effects of the present invention, human beings are preferable. In addition, in terms of further improving the effects of the present invention, among non-human animals, cattle, pigs, and horses are preferable, and pigs are more preferable.

[2. method for inhibiting the aging progression of ovary, ovarian follicle or ovum ]

Pyrroloquinoline quinone or a salt thereof has an effect of inhibiting the aging process of an ovary, a follicle or an ovum. Accordingly, as an embodiment of the present invention, there is provided a method for inhibiting the progression of senescence in an ovary, follicle or ovum, by ingesting a composition containing pyrroloquinoline quinone or a salt thereof. The ovary, follicle or ovum can be human ovary, follicle or ovum, or ovary, follicle or ovum of non-human animal such as cow, pig, horse, sheep, goat, dog, rabbit, mouse, rat, guinea pig, gerbil, hamster, ferret, etc. From the viewpoint of further improving the effect of the present invention, human ovaries, follicles, or ova are preferable. In addition, in terms of further improving the effect of the present invention, the ovary, follicle or ovum of the non-human animal is preferably the ovary, follicle or ovum of bovine, porcine or equine, and more preferably the ovary, follicle or ovum of porcine.

The composition according to these embodiments includes pyrroloquinoline quinone or a salt thereof in the types, amounts, daily intake amounts, and the like, other components in the types, amounts, daily intake amounts, and the like, and the form of the composition is as described in [1. anti-aging agent for ovary, follicle, or ovum ].

Examples

The present invention will be described in more detail below with reference to examples and the like. However, the present invention is not limited to the following examples. The following test examples were all carried out with the approval of the ethical committee for animal experiments at the university of Guangdong island.

[ test example 1: effect of administration of pyrroloquinoline quinone on AMH (anti-Mullerian hormone) in blood of female mice ]

2-week old lesch cell-specific NRG 1-depleted female mice were administered PQQ disodium salt (2 mg/kg/day) (n-5) by drinking water. In addition, only 2 weeks of water was administered to 6-month-old Lei's cell-specific NRG 1-deficient male mice (KO; control group) and 6-month-old wild-type mice (WT mice), respectively. Thereafter, blood was recovered, and the concentration of AMH (anti-mullerian hormone) as a follicle marker was determined using an AMH ELISA kit (Elabscience Biotechnology, Bethesda, MD) and according to the method of use of the kit. The results are shown in FIG. 1. In addition, mice with lacunae NRG 1-specific lewy cells had a short lifespan, corresponding to more than 40 years of age in humans at 6 months.

As shown in fig. 1, the concentration of AMH was decreased in the control group female mice to which no PQQ disodium salt was administered, compared to the WT mice, while the concentration of AMH was increased in the control group female mice to which PQQ disodium salt was administered, compared to the WT mice. That is, it was confirmed that the ovarian tissue recovered youth by the ingestion of PQQ disodium salt.

[ test example 2: effect of administration of pyrroloquinoline quinone on estrous cycle in female mice ]

PQQ disodium salt (2 mg/kg/day) was administered to 6-month old Lee's cell-specific NRG1 depleted female mice (KO) continuously by drinking water. In addition, female mice given only water were used as a control group. The oestrus cycle of each mouse was examined by vaginal smear examination. In addition, a vaginal smear test was performed as follows: the reflux was applied to the slide by vaginal reflux with physiological saline. After drying, fixation with methanol was performed and Giemsa staining (Giemsa staining) was performed. The slide glass was observed under an inverted microscope (manufactured by KEYENCE CORPORATION), and it was judged as a estrus when only leukocytes were observed, as a prophase of estrus when only nucleated epithelial cells were observed, and as an estrus when only non-nucleated epithelial cells were observed, and as a micro-estrus when a large number of non-nucleated epithelial cells were observed in the presence of non-nucleated epithelial cells and leukocytes, and as a postestrus when a large number of leukocytes were observed in the presence of non-nucleated epithelial cells and leukocytes. The results are shown in FIG. 2.

As shown in fig. 2, it can be confirmed that: while the female mice of the control group to which no PQQ disodium salt was administered maintained in a micro estrus state all the time and no periodic change in estrus was observed, the female mice to which PQQ disodium salt was administered began to estrus again after about 14 days from the initiation of administration. That is, it was confirmed that the abnormal ovulation cycle in female mice was improved by the administration of PQQ disodium salt.

[ test example 3: effect of administration of pyrroloquinoline quinone on ovum ]

Immature female mice of 3 weeks of age were administered PQQ disodium salt (2 mg/kg/day) (n-5) by drinking water. In addition, female mice given only water were used as a control group. At a time point after the start of administration and 2 days later, 4IU of eCG (equine chorionic gonadotropin) for inducing follicular development was administered, followed by 5IU of hCG (human chorionic gonadotropin) after 48 hours. Thereafter, the number of ovulations of each female mouse was determined. The number of ovulations was calculated by: the calculation was performed by recovering the oviduct from the killed mouse and incising the ampulla of the oviduct to recover the discharged ovum and cumulus cell complex and visually observing the complex under a stereo microscope. In addition, the discharged ovum was used for in vitro fertilization, and the fertilization rate and the development rate were calculated. Here, the fertilized egg refers to a split egg 24 hours after fertilization, and the fertilization rate refers to a ratio of the number of successfully fertilized eggs to the total number of discharged mature eggs. The development rate refers to the proportion of fertilized eggs that become blastocysts among the fertilized eggs. The results expressed as mean ± standard deviation for the number of ovulations and fertilization rate are shown in table 1. In addition, for mice, 3 weeks of age corresponds to the teens years of first menstruation in humans.

[ Table 1]

	Control group	PQQ disodium salt
			Number of ovulatory cycles	40.41±1.12	60.21±1.77
Fertilized egg number (number)	26.63±1.29	41.67±1.48
			Fertilization Rate (%)	65.77±2.10	69.41±0.77
Development Rate (%)	80.42±2.44	88.09±1.56

As shown in table 1, the number of ovulations of female mice administered with PQQ disodium salt was increased while maintaining the in vitro fertilization rate, and the development rate was increased while maintaining the in vitro fertilization rate, compared to the control female mice not administered with PQQ disodium salt. That is, it was confirmed that the egg quality was increased by the administration of PQQ disodium salt.

[ test example 4: effect of administration of pyrroloquinoline quinone on the concentration of estrogen in blood ]

Immature female mice of 3 weeks of age were administered PQQ disodium salt (2 mg/kg/day) by drinking water. In addition, female mice given only water were used as a control group. At a time point after the start of administration and 2 days later, 4IU of eCG (equine chorionic gonadotropin) for inducing follicular development was administered intraperitoneally, followed by 5IU of hCG (human chorionic gonadotropin) after 48 hours. Serum from each female mouse was recovered over time immediately before administration of eCG, and the blood estradiol (E2) concentration as a follicle development marker and progesterone (P4) concentration as a luteal marker were measured using a commercially available kit of Endocrine Technology, inc. (Newark, CA). The results are shown in FIG. 3.

As shown in fig. 3 (a), the blood estradiol concentration of the female mice administered with PQQ disodium salt increased with time compared to the control female mice not administered with PQQ disodium salt. That is, it was confirmed that the administration of PQQ disodium salt promoted the production of estradiol. It is known that estradiol is one of female hormones widely used in hormone replacement therapy, and is used for the treatment of climacteric disorder or the improvement of climacteric symptoms. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof can be used as an ameliorating or prophylactic agent for female symptoms caused by estrogen deficiency, or for female premenstrual syndrome or climacteric syndrome. Thus, the woman in the pre-menopause or the menopause can have amenorrhea without pain.

As shown in fig. 3 (B), no significant difference was observed between the female mice administered with PQQ disodium salt and the control group female mice not administered with PQQ disodium salt in terms of the increase in blood progesterone concentration. Progesterone is known to be associated with the concentration of blood in ovarian hyperstimulation syndrome. That is, it was confirmed that pyrroloquinoline quinone or a salt thereof can be used as a prophylactic agent for ovarian hyperstimulation syndrome by promoting the production of estradiol and relieving the production of progesterone.

[ test example 5: effect of administration of pyrroloquinoline quinone on ovarian fibrosis

6-month old Lee's cell-specific NRG1 depleted female mice (KO) were given 2 weeks PQQ disodium salt (2 mg/kg/day) by drinking water. Then, the removed ovaries were subjected to PSR staining and observed at a magnification of 20 times using an inverted microscope (manufactured by KEYENCE CORPORATION). The results are shown in FIG. 4.

As shown in fig. 4, the stroma cells of the mice were stained red overall before administration of PQQ disodium salt, and collagen deposition, i.e., ovarian fibrosis, was confirmed. In addition, atrophy of stromal cells was also observed. On the other hand, in the mice given 2 weeks PQQ disodium salt, it was confirmed that the red color of the stromal cells disappeared and the fibrotic stromal cells were improved to a healthy state. Stromal cells of the ovary are known to gradually fibrillate with increasing age. That is, it was confirmed that the ovarian tissue recovered youth by the ingestion of PQQ disodium salt. In addition, the follicular follicle was observed, and it was confirmed that atrophy of the follicle was also improved, and thus it was confirmed that the quality of the ovum was improved.

According to the results of test examples 1 to 5, by ingesting pyrroloquinoline quinone or a salt thereof, QOL of women who are easily impaired due to psychosomatic abnormalities occurring near amenorrhea can be ensured, and active and youth can be provided for elderly women as well.

Claims

1. An anti-aging agent for ovary, follicle or ovum, which comprises pyrroloquinoline quinone or a salt thereof.

2. An inhibitor or improver of fibrosis in female reproductive tissue, which comprises pyrroloquinoline quinone or a salt thereof.

3. An egg quality improving agent comprising pyrroloquinoline quinone or a salt thereof.

4. An ameliorating or prophylactic agent for menoxenia, which comprises pyrroloquinoline quinone or a salt thereof.

5. An ameliorating or prophylactic agent for ovarian hyperstimulation syndrome, which comprises pyrroloquinoline quinone or a salt thereof.

6. An estrogen production promoter contains pyrroloquinoline quinone or a salt thereof.

7. The estrogen production promoting agent according to claim 6, which improves or prevents symptoms of females caused by estrogen deficiency.

8. The estrogen production promoter according to claim 6, which improves or prevents a female from a pre-menopausal or climacteric disorder.

9. The estrogen production promoter according to claim 6, which improves or prevents female climacteric symptoms.

10. The formulation of any one of claims 1-9 for use in a non-human animal.

11. A method for inhibiting the aging process of an ovary, follicle or ovum by ingesting a composition comprising pyrroloquinoline quinone or a salt thereof.