JP2024028552A - Sperm fertility enhancer - Google Patents

Abstract

[Problem] To provide a sperm fertility enhancer. [Solution] A sperm fertility enhancer containing pyrroloquinoline quinone or its salt. [Selection diagram] None

Description

The present invention relates to a sperm fertility enhancer.

It is said that male infertility, where the main cause is on the male side, accounts for about 50% of all infertility cases. Examples of male infertility include oligozoospermia, in which sperm concentration in semen is low, and asthenozoospermia, in which sperm motility is poor. For male infertility, assisted reproductive treatments such as artificial insemination, in vitro fertilization, and microinsemination, which use sperm collected from semen, are applied (for example, Patent Document 1). However, such treatment imposes a heavy mental and physical burden on the patient, and is not economically easy.

Semen is composed of seminal plasma and sperm. Seminal plasma is a mixture of secretions from accessory gonads such as the seminal vesicles and the prostate gland. Sperm are not motile in the epididymis and begin to move when mixed with seminal plasma during ejaculation. Therefore, in order to increase fertility, not only the potential motility of sperm but also the function of seminal plasma itself is important. Note that sperm swim while vibrating their heads using flagella.

By the way, testosterone is a type of male hormone that is produced by Leydig cells in the testes and secreted into the blood. The concentration of testosterone in the testes is higher than in the blood, and this high concentration is essential for spermatogenesis. The main physiological effects of testosterone in adult men include promotion of spermatogenesis and protein anabolism in skeletal muscles. (Patent Documents 2 and 3)

In men, it is known that the amount of testosterone secreted decreases with age, and it is reported that 40% of all men over the age of 45 have a marked decrease in testosterone. This decrease in testosterone is said to originate from the failure of Leydig cells in the testis, and in male sexual function, it is associated with decreased libido, decreased spermatogenesis, andropause, and aging male hypogonadal syndrome (LOH syndrome). ), are known to increase the risk of prostate enlargement and obesity. These symptoms are also correlated with erectile dysfunction (ED), which is one of the sexual dysfunctions. In other words, the decrease in testosterone secretion due to aging results in a decrease in fertility, regardless of sperm motility. In this way, decreased testosterone secretion not only affects male infertility, but also causes symptoms of aging male hypogonadism syndrome, such as cognitive decline, hot flashes, depression, sleep disorders, muscle weakness, and glucose metabolism/lipidemia. It causes deterioration of metabolism, increase in visceral fat, osteoporosis, etc., and also significantly reduces the quality of life of men. Testosterone replacement therapy is known as a way to deal with this problem, but high-level clinical research has not yet been conducted and its effectiveness, risks and benefits are still being debated. It has been demanded. (Non-patent documents 1, 2, 3)

Decreased fertility due to decreased sperm motility is a problem not only in humans but also in livestock such as cows and pigs. For example, although artificial insemination is being popularized in the pig farming industry, it has been observed that there are many individuals whose sperm motility significantly decreases when semen is stored for a long period of time. Therefore, it is desired to develop a method for increasing fertility by increasing sperm motility not only for humans but also for non-human animals.

JP2012-228518A Japanese Patent Application Publication No. 2005-306754 JP2006-006311A

Journal of the Japanese Urological Association, 2004, Volume 95, p. 751 Sogo Rinbaku, 2011, Volume 60, p. 453 Treatment, 2017, Volume 99, p. 1132

An object of the present invention is to provide a sperm fertility enhancer.

The present inventors have discovered that pyrroloquinoline quinone or a salt thereof increases the rectilinear motility of sperm and also increases the fertility of sperm. The present invention is based on this new finding.

The present invention provides, for example, the following inventions.
[1]
A sperm fertility enhancer containing pyrroloquinoline quinone or its salt.
[2]
An agent containing pyrroloquinoline quinone or its salt, which suppresses decline in straight sperm motility.
[3]
A sperm rectilinear motility improving agent containing pyrroloquinoline quinone or its salt.
[4]
An accessory gonadal function improving agent containing pyrroloquinoline quinone or its salt.
[5]
A prostatic function improving agent or prostatic disease preventive agent containing pyrroloquinoline quinone or a salt thereof.
[6]
A seminal vesicle gland function improving agent containing pyrroloquinoline quinone or its salt.
[7]
An agent for improving or preventing aging male hypogonadism syndrome, which contains pyrroloquinoline quinone or a salt thereof.
[8]
A testosterone production promoter containing pyrroloquinoline quinone or its salt.
[9]
A spermatogenic ability improving agent containing pyrroloquinoline quinone or its salt.
[10]
An agent for improving or preventing male infertility, containing pyrroloquinoline quinone or a salt thereof.
[11]
An in vitro fertilization rate improving agent containing pyrroloquinoline quinone or its salt.
[12]
A method for increasing the fertility of sperm by ingesting a composition containing pyrroloquinoline quinone or its salt.
[13]
A method for suppressing a decrease in straight motility of sperm by ingesting a composition containing pyrroloquinoline quinone or a salt thereof.
[14]
A method for increasing the rectilinear motility of sperm by ingesting a composition containing pyrroloquinoline quinone or its salt.
[15]
A sperm preservation solution containing pyrroloquinoline quinone or its salt.
[16]
The sperm preservation solution according to [15], which is characterized by increasing the fertility of sperm.
[2-1]
The agent according to [1], which suppresses a decrease in sperm rectilinear motility.
[2-2]
The agent according to [1], which improves straight motility of sperm.
[2-3]
The agent according to [1], which promotes testosterone production.
[2-4]
The agent according to any one of [1], [8] and [2-3], which improves accessory gonadal function.
[2-5]
The agent according to any one of [8], [2-3] and [2-4], which improves prostate function or prevents prostate disease.
[2-6]
The agent according to any one of [8], [2-3] and [2-4], which improves seminal vesicle gland function.
[2-7]
The agent according to [8] or [2-3], which improves or prevents aging male hypogonadism syndrome.
[2-8]
The agent according to any one of [8] and [2-3] to [2-7], which improves spermatogenic ability.
[2-9]
The agent according to [1], which improves or prevents male infertility.
[2-10]
The agent according to [1], which improves in vitro fertilization rate.
[2-11]
The method according to [12], which is characterized by suppressing a decrease in straight motility of sperm.
[2-12]
The method according to [12], which is characterized by increasing the rectilinear motility of sperm.
[2-13]
Use of pyrroloquinoline quinone or a salt thereof in the manufacture of a composition for increasing sperm fertility.

According to the present invention, a sperm fertility enhancer can be provided.

2 is a graph showing the rectilinear motility of sperm when pyrroloquinoline quinone is added to the Hiroswine medium in Test Example 1. 2 is a graph showing the linear movement speed of sperm when pyrroloquinoline quinone is added to the Hiroswine medium in Test Example 1. 7 is a graph showing the testosterone concentration in each male mouse in Test Example 7. (A) and (B) are photographs taken of PSR-stained seminal vesicle gland cells of WT mice in Test Example 8. (C) and (D) are photographs of control (KO) seminal vesicle gland cells stained with PSR in Test Example 8. (E) and (F) are photographs of seminal vesicle gland cells of a mouse (KO) to which PQQ disodium salt was administered in Test Example 8, which were stained with PSR. Note that (A), (C), and (E) are images observed at 40x magnification, and (B), (D), and (F) are partially enlarged images of the 40x observation images. be.

EMBODIMENT OF THE INVENTION Hereinafter, the form for implementing this invention is demonstrated in detail. However, the present invention is not limited to the following embodiments.

[1. Sperm fertility enhancer]
The sperm fertility enhancer according to the present embodiment contains pyrroloquinoline quinone or a salt thereof (also referred to as "component (A)").

As used herein, "fertility of sperm" means the reproductive ability (fertility, fertility) of sperm. Fertility of sperm is affected by factors such as sperm concentration in semen, sperm motility, function of accessory gonads such as the prostate and seminal vesicles, and spermatogenic ability, resulting in decreased sperm concentration and lack of sperm motility. The fertility of sperm decreases due to decreased function of accessory gonads, decreased spermatogenic ability, decreased testosterone levels in the testes and blood, etc. Here, pyrroloquinoline quinone or a salt thereof has the effect of suppressing a decrease in the rectilinear motility of spermatozoa or increasing the rectilinear motility of spermatozoa. Therefore, as one embodiment of the present invention, there is provided an agent for suppressing a decrease in rectilinear sperm motility or an agent for improving rectilinear sperm motility, which contains pyrroloquinoline quinone or a salt thereof. Note that among motile spermatozoa, spermatozoa that move in a straight line have a high fertilization ability, and the higher the speed of the linear movement, the higher the fertilization ability.

In addition, pyrroloquinoline quinone or its salt has the effect of improving prostate function, seminal vesicle gland function, and accessory gonadal function, the effect of improving or preventing aging male hypogonadism syndrome, and the effect of preventing prostate disease. . Therefore, as one embodiment of the present invention, an agent for improving prostate function or preventing prostate disease, an agent for improving seminal vesicle gland function, an agent for improving accessory gonadal function, or an agent for improving gonadal function in aging men, which contains pyrroloquinoline quinone or a salt thereof, is provided. An agent for improving or preventing the syndrome is provided.

Furthermore, pyrroloquinoline quinone or a salt thereof has the effect of promoting the production of testosterone, the effect of improving spermatogenic ability, and the effect of improving the in vitro fertilization rate. Therefore, as an embodiment of the present invention, there is provided a testosterone production promoter, a spermatogenic ability improving agent, a male infertility improving agent or preventive agent, or an in vitro fertilization rate improving agent containing pyrroloquinoline quinone or a salt thereof. .

Therefore, pyrroloquinoline quinone or its salt, which has both of these effects, has the effect of increasing the fertility of sperm and the effect of improving or preventing male infertility. Therefore, as one embodiment of the present invention, there is provided an agent for enhancing sperm fertility, or an agent for improving or preventing male infertility, which contains pyrroloquinoline quinone or a salt thereof. Note that the agents containing pyrroloquinoline quinone or its salt according to each of the above embodiments are also collectively referred to as "the agent according to the present embodiment."

"Prostate function" includes, for example, secretion of prostatic fluid. Furthermore, examples of "prostate diseases" include tumorous diseases, inflammatory diseases, pyuria, and urinary disorders. Examples of neoplastic diseases include benign prostatic hyperplasia and prostate cancer. Examples of inflammatory diseases include acute prostatitis and chronic prostatitis. Symptoms of prostate disease include, for example, urinary dysfunction, hematuria, frequent urination, and bloody semen. The agent according to this embodiment can improve or prevent these symptoms by improving prostate function.

"Seminal vesicle gland function" includes, for example, secretion of seminal vesicle gland fluid. Further, examples of "seminal vesicle disease" include seminal vesicle adenitis, abnormal seminal vesicle dilatation, semen reflux disease, and semen leakage. Symptoms of seminal vesicle gland disease include, for example, urinary dysfunction, hematuria, frequent urination, hematosemen, and infertility. The agent according to this embodiment can improve or prevent these symptoms by improving seminal vesicle gland function.

Examples of "accessory gonads" include the prostate, seminal vesicles, epididymis, and bulbourethral glands. Examples of accessory gonadal functions include secretion of seminal plasma.

Symptoms of "Aging Male Hypogonadism Syndrome" include, for example, muscle weakness and associated muscle pain, hot flashes, sweating, headaches, dizziness, decreased sense of well-being, anxiety, and trivial symptoms. These symptoms include irritability, depression, insomnia, decreased concentration and memory accompanying these symptoms, decreased sexual desire (sexual arousal), and frequent urination. The agent according to the present embodiment can improve or prevent these symptoms by promoting the production of testosterone, and can serve as an alternative therapy to testosterone replacement therapy, which is expensive and whose safety and effectiveness are not sufficiently ensured. obtain. It is also known that blood flow improves when testosterone production is promoted. Blood flow disorders include arteriosclerosis, atherosclerosis, varicose veins, embolism, myocardial infarction, cerebral infarction, headache, hypertension, dementia, depression, renal failure, glomerulonephritis, ischemic hepatitis, acute pancreatitis, and ischemic bile duct. Piroro is a cause or aggravating factor for disorders such as Buerger's disease, Raynaud's disease, edema, joint pain, tinnitus, hearing loss (sensitivity hearing loss, noise-induced hearing loss), dizziness, Meniere's disease, hair loss, erectile dysfunction, and difficulty urinating. Quinoline quinone or a salt thereof has the effect of preventing or improving symptoms resulting from these blood flow disorders.

As used herein, "spermatogenic ability" means the ability to form healthy sperm, and specifically includes, for example, the ability to produce testosterone, the ability to improve the number of sperm in semen, and the ability to reduce the malformation rate of sperm. It will be done.

で表される公知の化合物である。 Pyrroloquinoline quinone has the following formula:

This is a known compound represented by

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

As the pyrroloquinoline quinone or its salt, 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 even more preferable.

Commercially available pyrroloquinoline quinone or its salt can also be used. Furthermore, as the pyrroloquinoline quinone or its salt, for example, those containing pyrroloquinoline quinone or its salt, such as natto, soybeans, cocoa powder, cacao mass, cacao, parsley, and green pepper, may be used.

When used in humans, the content of component (A) in the agent according to this embodiment is based on the total amount of the agent according to this embodiment, from the viewpoint of stability of the administered preparation, and pyrroloquinoline quinone or its salt. The total content of 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 even more preferably 1.3 to 3% Particularly preferred is weight %. Furthermore, when used in non-human animals, the content of component (A) in the agent according to this embodiment is such that the total content of pyrroloquinoline quinone or its salt is based on the total amount of the agent according to this embodiment. It is preferably 0.01 to 0.3% by weight, more preferably 0.02 to 0.2% by weight.

In the agent according to this embodiment, the daily intake amount of component (A) may vary depending on the condition of the ingesting individual (body weight, age, sex, etc.), the formulation form, etc., but when used in humans, From the viewpoint of ease of ingestion as a single dose, the effectiveness of the physiological action based on component (A), and the safety viewpoint, the amount is preferably 7 to 100 mg, more preferably 10 to 40 mg, and still more preferably 20 to 40 mg. When used in 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)"). Thereby, the effects of the present invention are more prominently exhibited.

Polyamines are aliphatic hydrocarbons containing two or more primary amino groups within the molecule. Specific examples of polyamines include putrescine, cadaverine, ethylenediamine, trimethylenediamine, hexamethylenediamine, spermidine, cardine, homospermidine, aminopropylcadaverine, spermine, theremine, thermospermine, canavarumine, aminopentylnorspermidine, and aminopropyl. Homospermine, canavarmine, homospermine, cardopentamine, homocardopentamine, aminopropylcanabarumine, bis(aminopropyl)homospermine, bis(aminopropyl)norspermine, aminobutylcanabarumine, aminopropylhomospermine, Examples include homopentamine, cardohexamine, homocardohexamine, sermohexamine, and homosermohexamine. As the polyamine, from the viewpoint of ease of use, spermidine, spermine, and putrescine are preferable, spermidine and spermine are more preferable, and spermidine is still more preferable.

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. In addition, as polyamines, extracts from plants such as soybeans (preferably soybean germ), wheat (preferably wheat germ), or rice (preferably rice germ), extracts from seafood (preferably milt), 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, from the viewpoint of ease of use, extracts from plants are preferred, and extracts from soybeans (preferably soybean germs) are more preferred. Commercially available polyamine compositions include, for example, "Soiporia" (manufactured by Combi Co., Ltd.), "Oryza Polyamine-P" (manufactured by Oryza Yuka Co., Ltd.), "Oryza Polyamine-LC" (manufactured by Oryza Yuka Co., Ltd.), Examples include "Phytopolyamine-S" (manufactured by Toyobo Co., Ltd.), "Phytopolyamine-SP" (manufactured by Toyobo Co., Ltd.), and "Elion SP" (manufactured by Mitsubishi Gas Chemical Co., Ltd.).

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

When the agent according to this embodiment contains a polyamine composition as component (B), the content of the polyamine composition is not particularly limited, and the type of polyamine composition, the type and content of other ingredients, and the preparation It is set as appropriate depending on the format etc. The content of the polyamine composition is determined from the viewpoint of the stability of the preparation, the effectiveness of physiological effects based on component (B), the viewpoint of safety, and the viewpoint of reducing the influence of the odor of component (B). For example, 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 based on the total amount of the agent according to the present embodiment. % is more preferable.

In the agent according to this embodiment, the content ratio of component (B) to component (A) is not particularly limited, and the types of component (A) and (B), the types and contents of other ingredients, and the formulation form. It is set as appropriate depending on the situation. From the viewpoint of further enhancing the effects of the present invention, the content ratio of component (B) to component (A) is, for example, per 1 part by weight of the total content of component (A) contained in the agent according to the present embodiment. The total content of 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 parts by weight. Parts by weight are more preferred.

In the case where the agent according to the present embodiment contains a polyamine composition as the (B) component, the content ratio of the polyamine composition to the (A) component is not particularly limited, and the type of the (A) component and the polyamine composition, etc. It is set as appropriate depending on the type and content of the ingredients, the formulation form, etc. From the viewpoint of further enhancing the effects of the present invention, the content ratio of the polyamine composition to component (A) is, for example, relative to 1 part by weight of the total content of 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 components (A) and (B) and other ingredients are It is set appropriately depending on the type, content, formulation form, etc. The weight ratio of spermine to spermidine is, for example, preferably from 0.01 to 10, more preferably from 0.1 to 2, and from 0.2 to 0. More preferably, it is .5.

In the agent according to this embodiment, the daily intake amount of component (B) may vary depending on the condition of the ingesting individual (body weight, age, sex, etc.), the formulation form, etc., but is based on the amount of 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 component (B), the amount is preferably 0.001 to 20 mg, more preferably 0.01 to 2 mg, and still more preferably 0.001 to 20 mg. 1 to 0.7 mg, particularly preferably 0.3 to 0.4 mg.

In the case where the agent according to this embodiment contains a polyamine composition as component (B), the daily intake amount of the polyamine composition depends on the condition of the ingesting individual (body weight, age, sex, etc.), the formulation form, etc. Although it may vary depending on the situation, it is preferably 50 to 400 mg, more preferably 100 to 300 mg, even more preferably 200 to 250 mg, particularly preferably 201 to 220 mg.

Crude medicines refer to all or part of natural products such as plants, animals, and minerals, either as they are or after simple processing such as drying, for the purpose of medicinal use. In the agent according to the present embodiment, the form of the crude drug includes, for example, the crude drug itself (crude drug); crude drug powder obtained by drying and powdering the crude drug; crude drug or crude drug powder mixed with water, organic solvents such as ethanol, or these. Examples include crude drug extracts extracted with mixed solvents, etc. Among these, herbal medicine extracts are preferred from the viewpoint of more significantly exerting the effects of the present invention. The crude drug extract may be the extract itself (tincture, liquid extract, etc.), the extract may be diluted or concentrated (soft extract, etc.), or the extract may be dried and then powdered. Alternatively, it may be in the form of a paste (such as a dried extract).

Specific examples of herbal medicine extracts include ginger extract, maca extract, licorice extract, Astragalus extract, chimpi extract, peony extract, Panax ginseng extract, turmeric extract, daisy extract, cinnamon bark extract, eleuthero extract, hawthorn extract, rhododendron extract, and Asian ginseng extract. , Saw Palmetto Extract, Ginkgo Biloba Extract, Saiko Extract, Citrus Extract, Sanyaku Extract, Sandalwood Extract, Sandalwood Extract, Soju Extract, Reishi Extract, Rokujo Extract, Tongkat Ali Extract, French Maritime Pine Bark Extract, European Oak Extract, Diosgenin Contains yam extract, cistanche extract, garlic extract, eleuthero extract, mucuna extract, gardenia extract, shilajit extract, cordyceps sinensis mycelium powder, black ginger extract, and black pepper extract. As crude drug extracts, maca extract, Panax ginseng extract, saw palmetto extract, ginger extract, Tongkat ali extract, French maritime pine bark extract, and European oak extract are preferable from the viewpoint of exhibiting the effects of the present invention more markedly, and maca extract and Panax ginseng extract is more preferable.

Commercially available crude drugs may be used. One type of crude drug may be used alone, or two or more types may be used in combination.

The content of component (C) in the agent according to the present embodiment is not particularly limited, and is appropriately set depending on the type of component (C), the type and content of other ingredients, the formulation form, etc. The content of component (C) may be, for example, from 0.1 to 0.1, based on the total amount of the agent according to the present embodiment, from the viewpoint of achieving more remarkable effects of the present invention. It is preferably 70% by weight, more preferably 1 to 10% by weight, even more preferably 2 to 4% by weight.

In the agent according to this embodiment, the content ratio of component (C) to component (A) is not particularly limited, and the types of component (A) and (C), the types and contents of other ingredients, and the formulation form It is set as appropriate depending on the situation. From the viewpoint of further enhancing the effect of the present invention, the content ratio of component (C) to component (A) is, for example, per 1 part by weight of the total content of component (A) contained in the agent according to the present embodiment. The total content of component (C) is preferably 0.1 to 3 parts by weight, more preferably 0.5 to 2.5 parts by weight, and 1 to 1.5 parts by weight. More preferably.

In the agent according to this embodiment, the content ratio of component (C) to component (B) is not particularly limited, and the types of component (B) and (C), the types and contents of other ingredients, and the formulation form It is set as appropriate depending on the situation. From the viewpoint of further enhancing the effects of the present invention, the content ratio of component (C) to component (B) is, for example, per 1 part by weight of the total content of component (B) contained in the agent according to the present embodiment. The total content of 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 can contain pharmacologically active ingredients or physiologically active ingredients 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 B6, vitamin B12, vitamin C, vitamin D, vitamin A, vitamin E, niacin, calcium pantothenate, taurine, Carnosine, anserine, balenine, citrulline, gamma-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, glutamic acid, glutamine, creatine, carnitine, luteolin, quercetin, genistin, cyanidin, resveratrol, diosgenin, isoflavone aglycone, lipo Examples include acid, zinc, iron, calcium, selenium, astaxanthin, zeaxanthin, β-carotene, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidonic acid, placenta extract, and pycnogenol. As the pharmacologically active ingredient or physiologically active ingredient, from the viewpoint of further enhancing the effect of the present invention, ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, and carnitine are preferable, and ubiquinone (coenzyme Q10), Astaxanthin and zinc are more preferred. One type of pharmacologically active ingredient or physiologically active ingredient may be used alone, or two or more types may be used in combination.

The agent according to the present embodiment can contain various additives in addition to the above-mentioned components, as long as they do not impair the effects of the present invention. Specific examples of such additives include excipients, binders, disintegrants, lubricants, colorants, flavoring agents, flavoring agents, sugars, sugar alcohols/polyhydric alcohols, and high-intensity sweeteners. Examples include oils, fats and oils, emulsifiers, thickeners, acidulants, fruit juices, etc. Excipients include lactose, sucrose, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, silicic acid, and the like. Examples of binders include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, calcium phosphate, polyvinylpyrrolidone, etc. Can be mentioned. Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, lactose, and the like. Examples of lubricants include talc, stearate, borax, polyethylene glycol, and the like. Examples of flavoring agents include white sugar, orange peel, citric acid, and tartaric acid. Examples of sugars include sugars such as sucrose, high fructose sugar, glucose, fructose, palatinose, trehalose, lactose, and xylose. Examples of sugar alcohols/polyhydric alcohols include sorbitol, xylitol, erythritol, lactitol, palatinit, reduced starch syrup, reduced maltose starch syrup, glycerin, propylene glycol, and the like. Examples of high-intensity sweeteners include aspartame, stevia, acesulfame potassium, and sucralose. Examples of the fats and oils include safflower oil, grape seed oil, sunflower oil, olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, and perilla oil. Examples of the emulsifier include sucrose fatty acid ester, glycerin fatty acid ester, lecithin, and the like. Examples of thickeners include carrageenan, xanthan gum, guar gum, pectin, and locust bean gum. Examples of acidulants include citric acid, lactic acid, and malic acid. Examples of fruit juices include lemon juice, orange juice, and berry juice. The additives may be used alone or in combination of two or more. From the viewpoint of stability of component (A) and component (B) during formulation, it is preferable that the additive contains oil or fat.

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

The agent according to this embodiment can be used, for example, as a component of pharmaceuticals, quasi-drugs, cosmetics, and food and beverages (beverages, foods). In addition, the agent according to the present embodiment is, for example, a pharmaceutical preparation, a quasi-drug preparation, a food for specified health use, a food with nutritional function, a food for the elderly, a food for special use, a food with functional claims, a health supplement (supplement), It can also be used in food preparations (eg confectionery tablets), obviously food products. Furthermore, the agent according to this embodiment can also be used as, for example, a veterinary drug, a feed additive, a diluent for frozen semen, and a diluent for artificial insemination.

Furthermore, when the agent according to the present embodiment is used as a food, the food may be a general food mixed with pyrroloquinoline quinone or its salt, and other ingredients as necessary. Preferable examples of such foods include those that can be stored for a long period of time, such as cookies, biscuits, snacks, jellies, gummies, chocolates, gums, candies, and cheese. It may also be a liquid food such as an energy drink, juice, tea drink, coffee drink, or milk drink.

The agent according to the present embodiment is suitable for use in subjects that require the effect of promoting sperm fertility (e.g., humans; cows, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters). , non-human animals such as ferrets). From the viewpoint of further enhancing the effects of the present invention, humans are preferred as subjects. In addition, from the viewpoint of further enhancing the effects of the present invention, among non-human animals, cows, pigs, and horses are preferable, and pigs are more preferable.

[2. A method for increasing the fertility of sperm, a method for suppressing a decline in sperm rectilinear motility, and a method for increasing sperm rectilinear motility]
Pyrroloquinoline quinone or a salt thereof has the effect of suppressing a decline in the rectilinear motility of sperm and the effect of increasing the rectilinear motility of sperm. Furthermore, pyrroloquinoline quinone or a salt thereof has the effect of increasing the fertility of sperm by suppressing a decrease in the rectilinear motility of sperm or increasing the rectilinear motility of sperm. Therefore, as one embodiment of the present invention, 1) a method for increasing the fertility of sperm, which involves ingesting a composition containing pyrroloquinoline quinone or a salt thereof; and 3) a method of suppressing a decline in sperm rectilinear motility by ingesting a composition containing pyrroloquinoline quinone or a salt thereof. The sperm may be human sperm or non-human animal sperm such as a cow, pig, horse, sheep, goat, dog, rabbit, mouse, rat, guinea pig, gerbil, hamster, or ferret. As spermatozoa, human spermatozoa are preferable from the viewpoint of further enhancing the effects of the present invention. Furthermore, from the viewpoint of further enhancing the effects of the present invention, among the non-human animal spermatozoa, bovine spermatozoa, porcine spermatozoa, and horse spermatozoa are preferred, and porcine spermatozoa are more preferred.

In addition, in these embodiments, the composition, such as the type, content and daily intake of pyrroloquinoline quinone or its salt, the type, content and daily intake of other components, etc. Regarding the formulation form etc. of the product, please refer to [1. As explained in [Sperm Fertility Enhancer].

[3. Sperm preservation solution]
Pyrroloquinoline quinone or its salt has the effect of increasing the fertility of sperm. Therefore, pyrroloquinoline quinone or its salt can also be used as a sperm preservation solution for preserving spermatozoa with increased fertility. Therefore, as one embodiment of the present invention, a sperm preservation solution containing pyrroloquinoline quinone or a salt thereof is provided.

The type of pyrroloquinoline quinone or its salt in the sperm preservation solution according to the present embodiment is [1. As explained in [Sperm Fertility Enhancer].

The content of pyrroloquinoline quinone or its salt in the sperm preservation solution according to the present embodiment is such that the total content of pyrroloquinoline quinone or its salt is 0.0000000033 to 0.00033% by weight based on the total amount of the sperm preservation solution. It is preferably 0.00000033 to 0.000033% by weight, and even more preferably 0.0000033% by weight.

The sperm preservation solution according to this embodiment may further contain at least one selected from the group consisting of lactose, galactose, sucrose, glycine, and lactic acid. Thereby, the effects of the present invention are more prominently exhibited.

The sperm preservation solution according to the present embodiment can contain pharmacologically active ingredients or physiologically active ingredients other than the above-mentioned ingredients, as long as the effects of the present invention are not impaired. Specific examples of such pharmacologically active ingredients or physiologically active ingredients include antibiotics such as amikacin, gentamicin, kanamycin, streptomycin, polymyxin B, and penicillin; and various amino acids. One type of pharmacologically active ingredient or physiologically active ingredient may be used alone, or two or more types may be used in combination.

The sperm preservation solution according to the present embodiment can contain various additives in addition to the above-mentioned components, as long as they do not impair the effects of the present invention. Specific examples of such additives include, for example, glucose; citric acid, carbonic acid, succinic acid, tartaric acid, malic acid, acetic acid, phosphoric acid or salts thereof; buffers such as tris(hydroxymethyl)aminomethane; ethylene diamine; Examples include chelating agents such as tetraacetic acid (EDTA) and glycol ether diamine tetraacetic acid (EGTA); cryoprotectants such as glycerin, ethylene glycol, propylene glycol, and dimethyl sulfoxide; and water. The additives may be used alone or in combination of two or more.

The sperm preservation solution according to this embodiment can be prepared by mixing pyrroloquinoline quinone or its salt and other components. Moreover, the sperm preservation solution according to this embodiment can also be prepared using a known preservation solution. For example, it can be adjusted by adding pyrroloquinoline quinone or its salt to a known storage solution. Known preservation solutions are not particularly limited as long as they are liquids commonly used in this field, but examples include Modena solution (components: glucose, sodium citrate, sodium hydrogen carbonate, citric acid, EDTA-2Na, tris(hydroxy) (methyl) aminomethane, penicillin and water). In this case, pyrroloquinoline quinone or its salt and, if necessary, other components may be added to the Modena liquid at a predetermined ratio and mixed.

The sperm preservation solution according to the present embodiment may contain human sperm, or non-human animals such as cows, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets (preferably cows, It can be used as a storage solution for spermatozoa (pig, horse). Moreover, the sperm preservation solution according to this embodiment can also be used, for example, as a diluent for frozen semen or a diluent for artificial insemination.

Hereinafter, the present invention will be explained more specifically 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: Analysis of linear motility and linear movement speed of spermatozoa]
In a 37°C water bath, add HIRO SWINE medium (HIRO SWINE B solution, manufactured by Cryo Preservation Service Co., Ltd.) containing various amounts of pyrroloquinoline quinone disodium salt (PQQ disodium salt) and PQQ disodium salt. Samples were prepared by suspending pig spermatozoa in untreated Hiroswine medium. Subsequently, the samples were incubated at 37°C for 2-6 hours. Next, 5 μL of the sample was analyzed using a computer aided sperm analysis (CASA) system. Regarding straight-line motility of sperm, the ratio of the number of straight-moving sperm among all observed spermatozoa is analyzed as a percentage, and the straight-line motion speed of sperm is determined by the average speed of straight-moving sperm. was analyzed. The results are shown in Figures 1 and 2.

As shown in FIG. 1, spermatozoa cultured in a medium to which PQQ disodium salt was added had improved rectilinear motility compared to spermatozoa to which PQQ disodium salt was not added. In addition, as shown in Figure 2, the average number of straight-moving spermatozoa for spermatozoa cultured in a medium supplemented with PQQ disodium salt was higher than that of spermatozoa cultured in a medium without PQQ disodium salt. Speed was improving. From the above, it was confirmed that the addition of PQQ disodium salt improves the linear motility and linear movement speed of sperm.

[Test Example 2: Effect of administration of pyrroloquinoline quinone on fertility of male mice]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, male mice to which only water was administered for 2 weeks were used as controls. Thereafter, the mice were allowed to live with female mice in heat for one hour to which hCG (placental gonadotropin) had been administered, and mating was attempted. This was done with one male and three females separately. After allowing the mice to coexist for 1 hour, mating was confirmed by the formation of a vaginal plug in the female mice, and the mating rate (%) was calculated (n = 3). Furthermore, 6 hours after the administration of hCG (placental gonadotropin), eggs were collected from the oviducts of female mice in which mating was confirmed, and the fertilization rate was measured. To determine fertilization, eggs in which two pronuclei had developed when observed under an inverted system microscope (IMT-2, manufactured by Olympus) were considered fertilized eggs. Fertilization rate was calculated using Equation 1 below.
(Formula 1) Fertilization rate (%) = {(Number of fertilized eggs)/(Total number of recovered eggs)} x 100
It is known that when NRG1 in Leydig cells is deficient, Leydig cell proliferation becomes impaired, blood testosterone concentration decreases, and spermatogenesis becomes abnormal. Furthermore, it is known that when NRG1 is deleted in Leydig cells, fertilization ability decreases. Furthermore, it is known that administering testosterone to NRG1-deficient mice increases the mating rate (Endoclinology, 157(12), p4899-4913, 2016). The results are shown in Table 1.

As shown in Table 1, in mice administered with PQQ disodium salt, both mating rate and fertilization rate increased compared to controls not administered with PQQ disodium salt, and the increase in mating rate was particularly remarkable. Ta. That is, it was confirmed that the administration of PQQ disodium salt enhanced the fertility of male mice and improved the decrease in libido (sexual arousal).

[Test Example 3: Effect of administration of pyrroloquinoline quinone on in vitro fertilization rate]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, male mice to which only water was administered for 2 weeks were used as controls. Thereafter, mature spermatozoa were collected from the epididymis of male mice and subjected to in vitro fertilization. Six hours after insemination, the eggs were collected and the in vitro fertilization rate (%) was measured. To determine fertilization, eggs in which two pronuclei had developed when observed under an inverted system microscope (IMT-2, manufactured by Olympus) were considered fertilized eggs. The results are shown in Table 2.

As shown in Table 2, the in vitro fertilization rate was increased in the sperm of mice administered with PQQ disodium salt compared to control sperm without administration of PQQ disodium salt. That is, it was confirmed that the administration of PQQ disodium salt enhanced the fertility of sperm.

[Test Example 4: Effect of administration of pyrroloquinoline quinone on spermatogenic ability]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, male mice to which only water was administered for 2 weeks were used as controls. Then, mature spermatozoa were collected from the epididymis of male mice, a sperm smear was prepared, fixed with methanol, and stained with hematoxylin and eosin. ) observed below. The determination of normal sperm or malformed sperm is based on the Nissan Gynecology (Volume 61, No. 6, Figure E-4-2-1 of N-189). Malformed spermatozoa were defined as abnormal sperm appearance, such as a circular sperm head, a bent sperm tail, or a missing sperm tail. 200 spermatozoa were observed for each individual, and the proportion of malformed spermatozoa among the 200 spermatozoa was calculated as the malformation rate (%) of the spermatozoa. The results are shown in Table 3.

As shown in Table 3, the malformation rate of the sperm of mice administered with PQQ disodium salt was significantly reduced to half that of control sperm without administration of PQQ disodium salt. That is, it was confirmed that administration of PQQ disodium salt improved sperm forming ability.

[Test Example 5: Effects of administration of pyrroloquinoline quinone on prostate function, seminal vesicle function, and accessory gonad function]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, male mice to which only water was administered for 2 weeks were used as controls. Thereafter, fluid components were collected from the accessory gonads (prostate, seminal vesicles, and epididymis) of male mice, and the fluid components were mixed. Sperm collected from the epididymis of a 6-month-old wild-type mouse (WT mouse) was cultured using the mixture, and the linear movement speed (μm/sec) of the sperm was measured after 2 hours for sperm motility analysis. The analysis was performed using a device (CASA).
By mixing fluid components from the prostate, seminal vesicles, and epididymis, it is possible to simulate seminal plasma. It is possible to reproduce semen. By analyzing the rectilinear motility of sperm at this time, it is possible to examine the effects on prostate function, seminal vesicle gland function, and accessory gonadal function. The results are shown in Table 4.

As shown in Table 4, the linear movement of spermatozoa cultured in seminal plasma of mice treated with PQQ disodium salt was significantly higher than that of control sperm cultured in seminal plasma of mice not treated with PQQ disodium salt. Improved speed. Therefore, it was confirmed that administration of PQQ disodium salt improves secretion of prostatic fluid, seminal vesicle gland fluid, and seminal plasma, that is, improves prostate function, seminal vesicle gland function, and accessory gonadal function.

[Test Example 6: Effect of administration of pyrroloquinoline quinone on seminal vesicle gland weight]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, male mice to which only water was administered for 2 weeks were used as controls. Thereafter, the seminal vesicle gland was removed and its weight was measured. The results are shown in Table 5.

As shown in Table 5, the weight of the seminal vesicle glands of mice administered with PQQ disodium salt increased compared to the seminal vesicle glands of controls to which PQQ disodium salt was not administered. That is, it was confirmed that seminal vesicle gland function was improved by administration of PQQ disodium salt.

[Test Example 7: Effect of administration of pyrroloquinoline quinone on testosterone concentration]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, water alone was administered to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO; control) and 6-month-old wild-type mice (WT mice) for 2 weeks, respectively. Thereafter, blood was collected from each mouse, followed by aliquoting 50 μl of serum. Testosterone was quantified using a Rodent testosterone ELISA kit (Endocrine technology; competitive method). The results are shown in Table 6 and Figure 3.

As shown in Table 6 and Figure 3, the testosterone concentration of mice administered with PQQ disodium salt was significantly improved compared to the testosterone concentration of controls not administered with PQQ disodium salt, and was comparable to that of normal mice. Testosterone levels improved to . That is, it was confirmed that administration of PQQ disodium salt promoted testosterone production and improved spermatogenic ability. A decrease in testosterone is a cause of andropause, aging male hypogonadal syndrome (LOH syndrome), and dysfunction of accessory gonads including the prostate, so it has also been confirmed that it can prevent or improve these symptoms. It was done.

[Test Example 8: Effect of administration of pyrroloquinoline quinone on seminal vesicle gland cells]
PQQ disodium salt was administered to drinking water (2 mg/kg/day) for 2 weeks to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO). In addition, water alone was administered to 6-month-old Leydig cell-specific NRG1-deficient male mice (KO; control) and 6-month-old wild-type mice (WT mice) for 2 weeks, respectively. Thereafter, the seminal vesicle glands were removed, fixed with paraformaldehyde, and then sections were obtained. This was stained with PAS and observed at 40x magnification using an inverted microscope (manufactured by Keyence). The results are shown in Figure 4. In addition, (A) and (B) are photographs taken of PSR-stained seminal vesicle gland cells of WT mice, and (C) and (D) are photographs of PSR-stained seminal vesicle gland cells of control (KO). (E) and (F) are photographs of PSR-stained seminal vesicle gland cells of a mouse (KO) administered with PQQ disodium salt. In addition, (A), (C), and (E) are images observed at 40x magnification, and (B), (D), and (F) are partially enlarged images of the 40x observation images. be.

As shown in Figure 4, in the control in which PQQ disodium salt was not administered, the seminal vesicle gland cells (represented by the cells shown in the circle in (D)) were small and atrophic, whereas PQQ disodium salt was not administered. The seminal vesicle gland cells (represented by the circled cells in (F)) of the administered mice improved to a size comparable to that of normal mice. In other words, it was confirmed that administration of PQQ disodium salt caused the atrophied seminal vesicle gland cells to recover, thereby improving seminal vesicle gland function.

From the results of Test Examples 1 to 8, it was confirmed that pyrroloquinoline quinone or its salt has the effect of enhancing the fertility of sperm. In other words, it was confirmed that it improves male infertility.

[Example of prescription for sperm preservation solution]
The components listed in Table 7 are mixed according to a conventional method to produce the sperm preservation solution of Formulation Example 1. The unit of each component in Table 7 is mg (milligram).

Claims (1)

Invention described in the specification.

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