JP7788095B2 - Preventive or therapeutic agent for nocturia - Google Patents

Description

The present invention relates to the prevention or treatment of nocturia.

Nocturia is a symptom in which a person must wake up at least once at night to urinate. The prevalence of nocturia increases with age, reaching an extremely high rate of 40-80% among the elderly. Furthermore, nocturia is a risk factor for sleep disorders, falls, and depression, and is associated with decreased quality of life and increased mortality. Therefore, improving nocturia could potentially improve quality of life and mortality in the elderly. Causes of nocturia include nocturia, bladder storage disorders, and sleep disorders, with nocturia being the most common cause. Nocturia is a condition in which urine volume increases during the night. The pathogenesis of nocturia is still poorly understood, and treatment methods have not yet been fully established. Desmopressin, the only treatment for nocturia, activates vasopressin V2 receptors in the renal collecting duct, reabsorbing water from urine and reducing urine volume. However, desmopressin has side effects, such as hyponatremia, and requires strict monitoring for side effects. Desmopressin also has limitations, such as its use in patients with hyponatremia or renal dysfunction, and its use in women. For this reason, desmopressin is not widely used as a treatment for nocturnal polyuria, and new treatments for nocturnal polyuria are needed.

Silicon microparticles can generate hydrogen when they come into contact with water. This reaction hardly progresses when they come into contact with an acidic aqueous solution with a pH of less than 5, but the reaction progresses when they come into contact with an aqueous solution with a pH of 7 or higher, and the reaction progresses more rapidly at a pH of 8 or higher. Surface-treating the silicon microparticles also facilitates this reaction. Furthermore, while the silicon microparticles are in contact with the aqueous solution, they continue to generate hydrogen for 20 hours or more, and under certain conditions, 1 gram of silicon microparticles can generate 400 ml or more of hydrogen (Patent Document 1, Patent Document 2, Non-Patent Document 1). 400 ml of hydrogen is equivalent to the hydrogen contained in 22 liters of saturated hydrogen water.

Patent Document 3 describes a solid preparation that contains silicon microparticles as its main ingredient and has the ability to generate hydrogen. However, there is no mention of the ability of silicon microparticles to prevent or treat disease.

Patent Document 4 describes a hydrogen supply material comprising a medium containing silicon microparticles and water. It also describes using this hydrogen supply material to supply hydrogen to the skin or mucous membranes. However, it does not describe how silicon microparticles can prevent or treat disease.

Patent Document 5 describes the treatment of silicon microparticles with hydrogen peroxide. However, it does not state that silicon microparticles can prevent or treat disease.

Patent Document 6 describes a formulation containing silicon microparticles, and cites embodiments in which the formulation is contained in a "matrix" such as animal medicines, livestock or pet food, animal feed, plant medicines, plant fertilizer, or plant compost. While it describes the formulation as promoting animal health and/or preventing disease, it does not state that the silicon microparticles are capable of preventing or treating disease to the extent that they could be used as a medicine.

Patent Document 7 primarily describes a silicon oxide film formed on the surface of silicon microparticles. It lists possible uses for the silicon microparticles, such as feed, supplements, food additives, and transdermal and/or transmucosal hydrogen uptake, and describes them as promoting animal health and/or preventing disease. However, it does not state that the silicon microparticles can prevent or treat disease to the extent that they could be used as a pharmaceutical.

Non-Patent Document 2 describes the effectiveness of hydrogen water for overactive bladder. Overactive bladder is a disease caused by excessive bladder contractions, and its main symptom is a sense of urgency to urinate, usually accompanied by frequent urination and nocturia. However, overactive bladder is a different pathology from nocturia, and Non-Patent Document 2 makes no mention of nocturia.

The applicant has filed patent applications for inventions relating to the use of silicon microparticles to prevent or treat kidney disease, inflammatory diseases (inflammatory bowel disease, arthritis, hepatitis, dermatitis), visceral discomfort, depression or depressive states, Parkinson's disease, autism spectrum disorder, memory impairment, spinal cord injury, hearing loss, cerebral ischemia-reperfusion injury, diabetes, and hangovers (Patent Documents 8 to 22).

A patent application for a preventive or therapeutic agent for diabetes (Patent Document 19) lists nocturia as one of the symptoms of diabetes. Nocturia associated with diabetes involves excessive urination throughout the day, not just at night, and is a different pathology from nocturnal polyuria.

Japanese Patent Application Laid-Open No. 2016-155118 Japanese Patent Application Laid-Open No. 2017-104848 International Publication No. 2017/130709 International Publication No. 2018/037752 International Publication No. 2018/037818 International Publication No. 2018/037819 International Publication No. 2019/211960 International Publication No. 2019/021769 International Publication No. 2019/235577 JP 2019-214556 A JP 2020-007300 A Japanese Patent Application Laid-Open No. 2020-079228 Japanese Patent Application Laid-Open No. 2020-079240 Japanese Patent Application Laid-Open No. 2020-117480 Japanese Patent Application Laid-Open No. 2020-117481 Japanese Patent Application Laid-Open No. 2020-117482 Japanese Patent Application Laid-Open No. 2020-117483 Japanese Patent Application Laid-Open No. 2020-117484 Japanese Patent Application Laid-Open No. 2020-117485 Japanese Patent Application Laid-Open No. 2020-117486 Japanese Patent Application Laid-Open No. 2020-200302 International Publication No. 2020/152985

Matsuda Shinsuke et al., Water decomposition and hydrogen concentration by silicon nanoparticles, Proceedings of the 62nd Spring Meeting of the Japan Society of Applied Physics, 2015, 11a-A27-6 Osamu Yamaguchi et al., Research Report for the Private University Strategic Research Infrastructure Formation Support Project from FY2014 to FY2018, pp. 250-253

The present invention aims to provide pharmaceuticals, medical devices, foods, beverages, etc. for the prevention or treatment of nocturia.

1. A preventive or therapeutic agent for nocturia containing silicon microparticles.
2. The preventive or therapeutic agent according to the preceding item 1, wherein the nocturia is caused by nocturnal polyuria.
3. The preventive or therapeutic agent according to the above item 1 or 2, which is for preventing or treating nocturia by regulating nocturnal urine volume.
4. The preventive or therapeutic agent according to any one of items 1 to 3 above, wherein the silicon microparticles are microparticles containing elemental silicon capable of generating hydrogen upon contact with water.
5. The preventive or therapeutic agent according to any one of items 1 to 4 above, wherein the silicon microparticles are silicon microparticles having a silicon oxide film formed on the surface thereof.
6. The preventive or therapeutic agent according to any one of items 1 to 5 above, wherein the silicon microparticles are silicon microparticles and/or aggregates of the silicon microparticles.
7. The preventive or therapeutic agent according to any one of items 1 to 5 above, wherein the silicon microparticles are porous silicon particles.
8. The preventive or therapeutic agent according to any one of items 1 to 7 above, wherein the silicon microparticles are silicon microparticles that have been hydrophilized.
9. The preventive or therapeutic agent according to any one of the preceding items 1 to 8, which is for oral administration.
10. A pharmaceutical composition for preventing or treating nocturia, containing silicon microparticles.
11. A medical device containing silicon microparticles for the prevention or treatment of nocturia.
12. A food or drink containing silicon microparticles for preventing or treating nocturia.
13. A therapeutic agent for nocturia containing silicon microparticles.
14. A method for preventing or treating nocturia, comprising administering silicon microparticles.
15. A method for treating nocturia comprising administering silicon microparticles.
16. An agent for use in the prevention or treatment of nocturia, containing silicon microparticles.
17. An agent for use in treating nocturia, containing silicon microparticles.
18. Use of silicon microparticles for the preparation of an agent for the prevention or treatment of nocturia.
19. Use of silicon microparticles for the preparation of a medicament for treating nocturia.

The preventive or therapeutic agent of the present invention can prevent and treat nocturia. The preventive or therapeutic agent of the present invention can preferably prevent and treat nocturnal polyuria. The preventive or therapeutic agent of the present invention can regulate nighttime urine volume. The preventive or therapeutic agent of the present invention is safe and has no side effects, and can be administered to many patients regardless of whether they suffer from other diseases, their medical history, gender, etc. Furthermore, it will contribute to improving the quality of life, maintaining their health, and improving the survival rate of many patients with nocturia.

FIG. 1 is a photograph of silicon microparticles (a mixture of silicon crystallites and their aggregates) taken with a scanning electron microscope (SEM) (Example 2). FIG. 2 is a graph showing the amount of hydrogen (cumulative amount) generated per 1 g of silicon microparticles by contacting the silicon microparticles obtained in Example 2 with water at 36° C. and pH 8.2. FIG. 3 is a photograph of silicon microparticles (aggregates of silicon crystallites) taken with a scanning electron microscope (SEM) (Example 3). Figure 4 shows the urine volume of old mice (80 weeks old) and young mice (19 weeks old) fed a high-salt diet. (a) shows the inactive phase urine volume rate, and (b) shows the daily urine volume. NS indicates mice fed a standard diet, and HS indicates mice fed a high-salt diet. Each group of 19-week-old mice: n = 6, each group of 80-week-old mice: n = 4, *p < 0.05, n.s.: not significant, Tukey-Kramer test Figure 5 shows the amount of nitrogen oxides (NOx) (NO ^2- , NO ^3- ) excreted daily by old mice (80 weeks old) and young mice (19 weeks old). 19-week-old mice: n=6, 80-week-old mice: n=4, *p<0.05, Student T-test Figure 6 shows the relationship between salt intake and nocturnal polyuria index in humans. (a) Low NOx group: n = 13 (b) High NOx group: n = 14, n.s.: not significant, Pearson product-moment correlation coefficient significance test Figure 7 shows the resting period urine output rate of nocturnal polyuria model mice. Control: n=6, nocturnal polyuria model: n=6, p<0.05, Student T-test 8 shows a method for examining the effect of silicon microparticles on nocturnal polyuria, where +Si indicates administration of silicon microparticles. Figure 9 is a graph showing the results of administering silicon microparticles to nocturnal polyuria model mice. The vertical axis shows the resting period urine volume rate. Control: n=6, Control + Si: n=7, Nocturnal polyuria model: n=6, Nocturnal polyuria model + Si: n=6; *p<0.05, Tukey-Kramer test 10A shows the results of Western blot analysis of NaCl cotransporter (NCC) and phosphorylated NCC (pNCC) in the kidneys of nocturnal polyuria model mice, and FIG. 10B shows a graph showing the ratio of each protein (pNCC/NCC). Nocturnal polyuria model: n=6, silicon microparticle-administered nocturnal polyuria model: n=6; p<0.05

The silicon microparticles contained in the preventive or therapeutic agent of the present invention are microparticles containing elemental silicon and can generate hydrogen when in contact with water.

The above-mentioned "microparticles containing elemental silicon capable of generating hydrogen upon contact with water" (silicon microparticles capable of generating hydrogen) refers to silicon microparticles that, when contacted with an aqueous solution at 36°C and pH 8.2, continuously generate hydrogen, generating 10 ml or more of hydrogen per gram of silicon microparticles in 24 hours. Preferably, this amount is 20 ml or more, 40 ml or more, 80 ml or more, 150 ml or more, 200 ml or more, or 300 ml or more.

The above-mentioned elemental silicon refers to high-purity silicon. In this specification, high-purity silicon refers to silicon with a purity of 98% or more, preferably 99% or more, more preferably 99.5% or more, more preferably 99.9% or more, more preferably 99.99% or more, and even more preferably 99.999% or more.

The silicon microparticles contained in the preventive or therapeutic agent of the present invention are preferably silicon microparticles, aggregates of such silicon microparticles, and/or porous silicon particles.

The active ingredient of the preventive or therapeutic agent of the present invention is preferably at least one type of particle selected from the group consisting of silicon microparticles, agglomerates of said silicon microparticles, and porous silicon particles. That is, the preferred active ingredient may be silicon microparticles alone, silicon microparticle agglomerates alone, or porous silicon particles alone. The active ingredient may also contain two or more types of silicon microparticles. The preventive or therapeutic agent of the present invention preferably contains silicon microparticles and/or agglomerates of said silicon microparticles. More preferably, the main ingredient is agglomerates of silicon microparticles.

When elemental silicon is exposed to the atmosphere, its surface is oxidized, forming a silicon oxide film. The silicon microparticles of the present invention are preferably microparticles having a silicon oxide film formed on their surface. Preferred silicon microparticles of the present invention are at least one type of particle selected from the group consisting of fine particles made of elemental silicon with a silicon oxide film formed on their surface, aggregates of such silicon microparticles, and porous silicon particles made of porous elemental silicon with a silicon oxide film formed on their surface.

The silicon content in the silicon microparticles is preferably 10% by weight or more, more preferably 20% by weight or more, even more preferably 50% by weight or more, and most preferably 70% by weight or more.

The silicon oxide film is preferably a silicon oxide film to which hydroxyl groups (-OH groups) have been added. A silicon oxide film to which hydroxyl groups have been added is a silicon oxide film that has been treated to increase the number of hydroxyl groups in the silicon oxide film. For example, hydroxyl groups can be added to the silicon oxide film by hydrophilization treatment. Silicon microparticles formed with a silicon oxide film to which hydroxyl groups have been added have improved contact efficiency between the surface and water, promoting the hydrogen generation reaction and enabling the generation of large amounts of hydrogen. The hydrophilization method is not particularly limited, and any known hydrophilization method can be used. Examples include hydrogen peroxide treatment and nitric acid treatment. Hydrogen peroxide treatment is preferred. Hydrogen peroxide treatment removes hydrogen from the SiH groups in the silicon oxide film on the particle surface, allowing hydroxyl groups to be added to the particle surface.

The silicon microparticles having a silicon oxide film with hydroxyl groups added thereto on their surfaces preferably have 5×10 ¹³ /cm ² or more hydroxyl groups on their surfaces. More preferably, they have 1×10 ¹⁴ /cm ² or more hydroxyl groups. Even more preferably, they have 3×10 ¹⁴ /cm ² or more hydroxyl groups. The particle surfaces include the surfaces of silicon microparticles, the surfaces of porous silicon particles, the surfaces of aggregates of silicon microparticles, and the surfaces of silicon microparticles that form aggregates.

Specific methods for hydrogen peroxide treatment include immersing silicon microparticles in hydrogen peroxide and stirring them. The hydrogen peroxide concentration is preferably 1-30%, more preferably 1.5-20%, even more preferably 2-15%, 2.5-10%, and most preferably 3-5%. The immersion and stirring time is preferably 5-90 minutes, more preferably 10-80 minutes, and even more preferably 20-70 minutes. Most preferably 30-60 minutes. Treatment with hydrogen peroxide can improve the hydrophilicity of silicon microparticles, but longer treatment times will cause the hydrogen generation reaction from the silicon microparticles to progress, increasing the thickness of the oxide film on the silicon microparticles. The temperature of the hydrogen peroxide during hydrogen peroxide treatment is preferably 20-60°C, more preferably 25-50°C, even more preferably 30-40°C, and most preferably 35°C.

There are no restrictions on the shape of the silicon microparticles. Examples include irregular, polygonal, spherical, elliptical, cylindrical, etc.

The silicon microparticles may be crystalline silicon microparticles, which have crystallinity. They may also be amorphous silicon microparticles, which do not have crystallinity. If they have crystallinity, they may be single crystal or polycrystalline. Crystalline silicon microparticles are preferred, and single crystal silicon microparticles are more preferred.

The amorphous silicon microparticles may be amorphous silicon microparticles formed by a method such as plasma CVD or laser ablation.

The silicon oxide film formed on the surface of silicon microparticles in the present invention may be a silicon oxide film formed by natural oxidation upon exposure to the atmosphere. It may also be a silicon oxide film formed artificially by known methods such as chemical oxidation using an oxidizing agent such as nitric acid.

The thickness of the silicon oxide film may be any thickness that stabilizes the particles made of elemental silicon and enables efficient hydrogen generation. For example, the thickness may be 0.3 nm to 5 nm, 0.3 nm to 3 nm, 0.5 nm to 2.5 nm, 0.7 nm to 2 nm, 0.8 nm to 1.8 nm, or 1.0 nm to 1.7 nm. The silicon oxide film may be a film containing oxides such as _SiO , SiO, _SiO3 , _{and SiO2} that are formed when silicon on the surface of the particles made of elemental silicon combines with oxygen. Oxides formed by incomplete oxidation of silicon, such as _SiO , SiO, and _{SiO3 ,} promote the hydrogen generation reaction.

The silicon microparticles may be crystalline silicon microparticles, which have crystallinity. They may also be amorphous silicon microparticles, which do not have crystallinity. If they have crystallinity, they may be single crystal or polycrystalline. Preferred silicon microparticles are crystalline silicon microparticles, and more preferred are single crystal silicon microparticles (hereinafter also referred to as silicon crystallites).

The silicon microparticles of the present invention may be silicon microparticles on which a silicon oxide film has been formed naturally or artificially after production. More preferred silicon microparticles are those on which a silicon oxide film has been formed on the surface of the silicon crystallites.

The silicon microparticles in the present invention may be particles obtained by crushing lumps of elemental silicon (high-purity silicon) or particles of elemental silicon. When elemental silicon lumps or particles are crushed to produce silicon microparticles, the surfaces of the silicon microparticles naturally oxidize to form a silicon oxide film.

The particle diameter of the silicon microparticles in the present invention (crystallite diameter when the microparticles are silicon crystallites) is preferably 0.5 nm or more and 100 μm or less, more preferably 1 nm or more and 50 μm or less, more preferably 1.5 nm or more and 10 μm or less, more preferably 2 nm or more and 5 μm or less, more preferably 2.5 nm or more and 1 μm or less, 5 nm or more and 500 nm or less, 7.5 nm or more and 200 nm or less, or 10 nm or more and 100 nm or less. A particle diameter of 500 nm or less provides a suitable rate and amount of hydrogen generation, and a particle diameter of 200 nm or less provides an even more suitable rate and amount of hydrogen generation.

The silicon microparticle aggregates of the present invention are aggregates of the silicon microparticles described above. They may be naturally formed or artificially formed. Preferably, they are aggregates of silicon microparticles on which a silicon oxide film is formed. Naturally formed aggregates are thought to remain aggregated in the digestive tract. Preferred aggregates have internal voids that allow water molecules to penetrate the aggregate and react with the microparticles inside. The rate of hydrogen generation from naturally formed aggregates does not depend on the aggregate size, so the aggregates have a structure that allows internal voids that allow water molecules to penetrate the aggregate and react with the microparticles inside.

There are no particular restrictions on the size of the silicon microparticle agglomerates. The preferred particle size of the silicon microparticle agglomerates is 50 nm or more and 500 μm or less.

The particle diameter of the silicon microparticles constituting the silicon microparticle agglomerates in the present invention is preferably 0.5 nm to 100 μm, more preferably 1 nm to 50 μm, more preferably 1.5 nm to 10 μm, more preferably 2 nm to 5 μm, more preferably 2.5 nm to 1 μm, 5 nm to 500 nm, 7.5 nm to 200 nm, or 10 nm to 100 nm. The silicon microparticles constituting the silicon aggregates may be crystalline silicon microparticles or amorphous silicon microparticles. Preferred aggregates are aggregates of silicon crystallites with a crystallite diameter of 1 nm to 10 μm. Preferably, the aggregates are formed by agglomerating silicon crystallites having a silicon oxide film formed on their surfaces.

The preventive or therapeutic agent of the present invention preferably contains silicon crystallites with a crystallite diameter of 1 nm to 1 μm, more preferably 1 nm to 100 nm, with a silicon oxide film formed on their surface, and/or aggregates thereof. Preferably, the agent contains, as a main component, aggregates of silicon crystallites with a silicon oxide film formed on their surface.

The preventive or therapeutic agent of the present invention preferably contains silicon crystallites with a crystallite diameter of 1 nm to 1 μm, more preferably 1 nm to 100 nm, and contains crystallites and/or aggregates thereof, on whose surface a silicon oxide film with added hydroxyl groups is formed. Preferably, the agent contains, as a main component, aggregates of silicon crystallites on whose surface a silicon oxide film with added hydroxyl groups is formed.

Porous silicon particles can be a porous body of silicon particles. They can also be a porous body formed by agglomerating and processing fine silicon particles. The porous silicon particles are preferably particles made of porous silicon alone, with a silicon oxide film formed on the surface. More preferably, the silicon oxide film is a silicon oxide film with hydroxyl groups added.

The porous silicon particles may be crystalline porous silicon particles. Alternatively, they may be amorphous porous silicon particles that do not have crystallinity. If they have crystallinity, they may be single crystal or polycrystalline.

There is no limit to the size of the voids present in the porous silicon particles, but they can typically be between 0.3 nm and 1 μm, and preferably between 0.5 nm and 0.1 μm. The porous silicon particles have a sufficient surface area to achieve high hydrogen generation capacity. The porous silicon particles may aggregate to form agglomerates. There are no particular limits to the size of the porous silicon particles or the aggregates of porous silicon particles. The size can preferably be between 200 nm and 400 μm.

Agglomerates of silicon microparticles and porous silicon particles have a large overall particle size and a large surface area, making them ideal for oral administration. Larger particles do not pass through the cell membranes and spaces between cells in the digestive tract, particularly the intestinal tract, and the silicon microparticles are not absorbed into the body, making them superior from a safety perspective.

There are no particular restrictions on the particle size distribution of the silicon microparticles contained in the preventive or therapeutic agent of the present invention, or on the particle size distribution or crystallite size distribution of the microparticles consisting of simple silicon. They may be polydisperse. They may also be formulations containing silicon microparticles with a specific range of particle size or crystallite size. Furthermore, there are no particular restrictions on the size distribution of aggregates of the silicon microparticles.

The rate at which hydrogen is generated can be adjusted by the particle size and particle size distribution of the silicon microparticles and/or the thickness of the silicon oxide film.

The silicon microparticles of the present invention can be produced by physically pulverizing silicon-containing particles to the desired particle size without any particular limitations. Suitable physical pulverization methods include bead mill pulverization, planetary ball mill pulverization, shock wave pulverization, high-pressure collision pulverization, jet mill pulverization, or a combination of two or more of these. Known chemical methods can also be used. From the perspective of production cost and ease of production management, physical pulverization is the preferred pulverization method. When microparticles consisting of fine particles of elemental silicon are exposed to the atmosphere, their surfaces are oxidized, forming a silicon oxide film. Alternatively, a silicon oxide film can be artificially formed after pulverization by known methods such as chemical oxidation using an oxidizing agent such as hydrogen peroxide or nitric acid.

When silicon-containing particles are produced by grinding them to the desired particle size using a bead mill, the desired particle size or particle size distribution can be obtained by appropriately changing the size and/or type of beads.

There are no restrictions on the silicon-containing particles used as the starting material, as long as they are high-purity silicon particles. For example, commercially available high-purity silicon particle powders can be used. The silicon-containing particles used as the starting material can be single-crystalline, polycrystalline, or amorphous.

This application includes inventions relating to an agent for preventing or treating nocturia containing silicon microparticles, an invention relating to a method for preventing or treating nocturia including administering silicon microparticles, an invention relating to an agent for use in preventing or treating nocturia containing silicon microparticles, and an invention relating to the use of silicon microparticles for preparing an agent for preventing or treating nocturia. The description and embodiments of the invention relating to an agent for preventing or treating nocturia containing silicon microparticles in this specification include description and embodiments of all of these inventions.

The preventive or therapeutic agent for nocturia of the present invention includes an agent for preventing nocturia, an agent for treating nocturia, and an agent for both preventing and treating nocturia.

As used herein, nocturia refers to the condition of having to wake up at least once at night to urinate. The preventive or therapeutic agent of the present invention has the effects of preventing the onset of nocturia, delaying the onset, improving symptoms, suppressing the worsening of symptoms, preventing the recurrence of symptoms, and promoting early recovery from symptoms.

The preventive or therapeutic agent of the present invention is preferably capable of preventing and treating nocturnal polyuria caused by nocturnal polyuria. As used herein, nocturnal polyuria refers to a condition in which a large proportion of the daily (24-hour) urine volume occurs at night. Nocturnal urine volume does not include the last urine voided before going to bed, but does include the first urine voided after waking up. Nocturnal polyuria is generally defined as a condition in which the nocturnal polyuria index (the ratio of nighttime urine volume to daily urine volume) is 33% or higher in people aged 65 or older, and 20% or higher in young adults.

The preventive or therapeutic agent of the present invention can regulate nocturnal urine volume, and the present invention can also be referred to as a urine volume regulator containing silicon microparticles. Preferably, the present invention can be referred to as a nocturnal urine volume regulator.

The preventive or therapeutic agent of the present invention can prevent and treat nocturia by adjusting nighttime urine volume and lowering the nocturnal polyuria index. In this specification, adjusting nighttime urine volume means reducing nighttime urine volume or lowering the nocturnal polyuria index. The preventive or therapeutic agent of the present invention can reduce nighttime urine volume without significantly changing daily urine volume.

The silicon microparticles of the present invention have the property of continuing to generate hydrogen for long periods of time (20 hours or more) in vitro. The silicon microparticles of the present invention generate hydrogen when they come into contact with water at a pH of 7 or higher, and generate even more hydrogen at a pH of 8 or higher. On the other hand, they have the property of generating almost no hydrogen at a pH of 5 or lower.

When the silicon microparticles of the present invention are orally administered, due to the properties described above, it is thought that little hydrogen is generated in the stomach, but hydrogen is generated in the intestines. When the silicon microparticles of the present invention were administered to normal mice, hydrogen generation was confirmed in the cecum, a part of the large intestine. Even when normal mice were fed a normal diet under the same conditions, hydrogen was below the detection limit. Since the retention time of food in the intestines is typically 20 hours or more in humans, it is thought that the preventive or therapeutic agent of the present invention, when administered orally, continues to generate hydrogen in the intestines for a long period of time, allowing hydrogen to be distributed throughout the body.

It is also believed that by leaving silicon microparticles on the skin or mucous membranes for an extended period of time, hydrogen can be delivered transdermally or transmucosally throughout the body for an extended period of time.

Furthermore, unlike hydrogen water, the preventive or therapeutic agent of the present invention does not allow hydrogen to diffuse before administration. This property contributes to maintaining the quality of pharmaceuticals and other products, and contributes to convenience for manufacturers, sellers, and users.

The inventors' research has confirmed that when rats were administered the silicon microparticles of the present invention and then their plasma antioxidant capacity was evaluated (BAP test), the antioxidant capacity was significantly higher in the group administered the silicon microparticles (WO2019/235577).

The mechanism by which the silicon microparticles of the present invention are effective in preventing or treating nocturia is currently under research, but it is thought to involve the inhibition of phosphorylation of NCC (NaCl cotransporter) in the distal renal tubules. It is also thought that a reduction in oxidative stress may also be involved.

Regarding the oxidative stress reduction effect, research by the present inventors and others has confirmed that silicon nanoparticles exhibit a significant effect compared to hydrogen water in studies using disease model animals involving oxidative stress (WO 2019/235577). Therefore, it is conceivable that silicon nanoparticles may have other effects not found in hydrogen water. Comparing the colonic tissue of mice administered with and without silicon nanoparticles, the colonic tissue of mice administered with silicon nanoparticles contained higher amounts of glutathione monosulfide and cysteine monosulfide, which are involved in antioxidant effects in vivo. This may be a unique effect of silicon nanoparticles (JP 2020-117481, PCT/JP2021/016176). Other possible mechanisms include, for example, proteins containing metal elements such as cobalt capturing early-stage hydrogen generated in the intestine by the reaction of silicon nanoparticles with water, or proteins with increased reducing power as a result of hydrogen atoms donating electrons, being transported to various organs and reacting with hydroxyl radicals to eliminate them.

The subjects for prevention or treatment with the prophylactic or therapeutic agent of the present invention are humans and non-human animals. Preferred non-human animals include pets and livestock.

One or more types of silicon microparticles of the present invention may be administered directly to humans or non-human animals. However, if necessary, they may be mixed with acceptable additives or carriers and formulated into forms known to those skilled in the art. Examples of such additives or carriers include pH adjusters (e.g., sodium bicarbonate, sodium carbonate, potassium carbonate, citric acid, etc.), excipients (e.g., sugar derivatives such as mannitol and sorbitol; starch derivatives such as corn starch and potato starch; or cellulose derivatives such as crystalline cellulose), lubricants (e.g., metal stearates such as magnesium stearate; or talc), binders (e.g., hydroxypropyl cellulose, hydroxypropylmethylcellulose, or polyvinylpyrrolidone), disintegrants (e.g., cellulose derivatives such as carboxymethylcellulose and calcium carboxymethylcellulose), and preservatives (e.g., parahydroxybenzoates such as methylparaben and propylparaben; or alcohols such as chlorobutanol and benzyl alcohol). These additives and carriers can be blended into the silicon microparticles either alone or in combination of two or more. Preferred additives include pH adjusters capable of adjusting the pH to 8 or higher. Preferred pH adjusters include sodium bicarbonate.

There are no particular limitations on the route of administration of the preventive or therapeutic agent of the present invention, but preferred routes of administration include oral, transdermal, and transmucosal (oral, rectal, vaginal, etc.) administration.

Formulations for oral administration include tablets, capsules, granules, powders, syrups (dry syrups), and oral jellies. Formulations for transdermal or transmucosal administration include patches and ointments.

Tablets, capsules, granules, powders, and the like can be made into enteric-coated preparations. For example, tablets, granules, and powders can be coated with an enteric coating. The enteric coating agent can be a gastric-insoluble enteric coating agent. Capsules can be made enteric-coated by filling enteric capsules with the silicon microparticles of the present invention.

The preventive or therapeutic agent of the present invention can be formulated into the above-mentioned dosage form and then administered to humans or non-human animals.

The content of silicon microparticles in the preventive or therapeutic agent of the present invention is not particularly limited, but examples include 0.1 to 100% by weight, 1 to 99% by weight, and 5 to 95%.

The dosage and frequency of administration of silicon microparticles in the present invention can vary appropriately depending on factors such as the recipient, their age, weight, sex, purpose (e.g., prevention or treatment), severity of symptoms, dosage form, and route of administration. When administered to humans, a preferred dosage of silicon microparticles is, for example, approximately 10 mg to 10 g per day, preferably approximately 100 mg to 5 g, and more preferably approximately 500 mg to 2 g. Furthermore, the frequency of administration may be one or more times per day, or once every few days. For example, it may be one to three times, one to two times, or once per day.

The agent for preventing or treating nocturia containing the silicon microparticles of the present invention can be used in pharmaceuticals, quasi-drugs, medical devices, foods, and beverages.

The present application also relates to an invention of a pharmaceutical composition for preventing or treating nocturia, which contains the silicon microparticles. The present application also relates to an invention of a pharmaceutical composition for preventing or treating nocturia, which contains the silicon microparticles as a preventive or therapeutic agent for nocturia. The pharmaceutical composition of the present invention also includes compositions with mild effects that fall under the category of quasi-drugs. Embodiments of the pharmaceutical composition of the present invention include embodiments of the invention relating to the preventive or therapeutic agent described above.

This application also relates to an invention for a medical device for preventing or treating nocturia, which contains the agent for preventing or treating nocturia containing the silicon microparticles. This application also relates to an invention for a medical device for preventing or treating nocturia, which contains the silicon microparticles. A medical device in this invention is an instrument or device intended for use in the treatment or prevention of disease in humans or non-human animals. An example of a medical device is a mask. By wearing the mask of the present invention, hydrogen can be supplied directly to the trachea or lungs. Another example is a bandage.

The present application also relates to an invention for a food or beverage for preventing or treating nocturia, which contains the agent for preventing or treating nocturia containing the silicon microparticles. The present application also relates to an invention for a food or beverage for preventing or treating nocturia, which contains the silicon microparticles. Preferred examples of the food or beverage of the present invention include health foods, functional foods, and foods for specified health uses. These health foods, functional foods, and foods for specified health uses are foods or beverages that can prevent the onset of nocturia symptoms, delay the onset, and/or prevent the recurrence of symptoms. There are no limitations on the form of the food or beverage. For example, they may be in the form of a mixture mixed with an existing food or beverage, or in the form of a formulation. Examples include tablets, capsules, powders, granules, jellies, etc.

The present invention will be explained in more detail below using examples, but the present invention is not limited to these examples.

Example 1
High-purity silicon powder (Osaka Titanium Technologies Co., Ltd., particle size distribution <φ300 μm (however, silicon particles with a crystal particle diameter of more than 1 μm), purity 99.9%) was sieved to remove particles of 45 μm or larger. 200 g of the obtained silicon particles were dispersed in 4 L (liters) of 99.5 wt % ethanol solution, and φ0.5 μm zirconia beads (volume 750 ml) were added. The mixture was then pulverized (single-stage pulverization) for 4 hours at 2500 rpm using a bead mill (Imex Co., Ltd., horizontal continuous ready mill (model RHM-08)).

The ethanol solution containing the micronized silicon particles was separated from the beads using a separation slit installed inside the grinding chamber of the bead mill, and then heated to 30-35°C using a vacuum evaporator. Micronized silicon particles (crystallites) were obtained by evaporating the ethanol solution.

The average crystallite diameter of the micronized silicon particles (crystallites) obtained by the above method was 20 to 30 nm, and most of the crystallites formed aggregates. Furthermore, the crystallites were coated with a silicon oxide film, which was approximately 1 nm thick. The resulting mixture of silicon crystallites coated with a silicon oxide film and their aggregates is one embodiment of the silicon microparticles that are the active ingredient of the present invention.

Example 2
The silicon crystallites and their aggregates obtained in Example 1 were mixed with hydrogen peroxide (3 wt%) in a glass container and stirred at 35°C for 30 minutes. The silicon crystallites and their aggregates treated with hydrogen peroxide were subjected to solid-liquid separation using a known centrifugal separator to remove the hydrogen peroxide. The obtained silicon crystallites and their aggregates were then mixed with an ethanol solution (99.5 wt%) and thoroughly stirred. The silicon crystallites and their aggregates mixed with the ethanol solution were subjected to solid-liquid separation using a known centrifugal separator to remove the highly volatile ethanol solution, and then thoroughly dried. The obtained mixture of silicon crystallites and their aggregates treated with hydrogen peroxide and having a silicon oxide film formed thereon is one embodiment of the silicon microparticles that are the active ingredient of the present invention. A scanning electron microscope (SEM) photograph of the obtained silicon microparticles is shown in Figure 1. The hydrogen generation rate of the obtained silicon crystallite aggregates did not depend on the aggregate size.

The amount of hydrogen generated from the silicon microparticles (silicon crystallites and their aggregates) obtained in Example 2 was measured. 10 mg of silicon microparticles were placed in a 100 ml glass bottle (borosilicate glass, approximately 1 mm thick, ASONE Laboran screw cap bottle). Water adjusted to pH 8.2 with sodium bicarbonate was added to the bottle, and the bottle was sealed at a temperature of 36°C. The hydrogen concentration in the liquid inside the bottle was measured. A portable dissolved hydrogen meter (Toa DKK Corporation, Model DH-35A) was used to measure the hydrogen concentration. The amount of hydrogen generated per 1 g of silicon microparticles is shown in Figure 2.

Example 3
In the same manner as in Example 2, the silicon microparticles (silicon crystallites and their aggregates) obtained in Example 1 were treated with hydrogen peroxide, mixed with an ethanol solution, and stirred. The silicon microparticles mixed with the ethanol solution were dried using a spray dryer (ADL311S-A, manufactured by Yamato Scientific). The obtained silicon crystallite aggregates are one embodiment of the silicon microparticles that are the active ingredient of the present invention. A scanning electron microscope (SEM) photograph of the obtained silicon microparticles (silicon crystallite aggregates) is shown in Figure 3.

Example 4
Single-stage pulverization was carried out in the same manner as in Example 1. The φ0.5 μm zirconia beads (volume 750 ml) used in the single-stage pulverization were automatically separated from the solution containing silicon crystallites in the bead mill pulverization chamber. 0.3 μm zirconia beads (volume 750 ml) were added to the resulting solution containing silicon crystallites, and the silicon crystallites were further pulverized (two-stage pulverization) at a rotation speed of 2500 rpm for 4 hours to refine them.

The beads were separated from the solution containing the silicon crystallites as described above, and the resulting ethanol solution containing the silicon crystallites was heated to 40°C using a vacuum evaporator, as in Example 1. The ethanol evaporated, and two-stage crushed silicon crystallites were obtained. These two-stage crushed silicon crystallites bearing a silicon oxide film are also one embodiment of the silicon microparticles that are the active ingredient of the present invention.

Example 5
The mixture of silicon crystallites and their aggregates formed with a silicon oxide film treated with hydrogen peroxide obtained in Example 2 was filled into commercially available capsules No. 3 to obtain a capsule preparation. This capsule preparation contains, as its main component, aggregates of silicon crystallites formed with a silicon oxide film treated with hydrogen peroxide, and further contains silicon crystallites formed with a silicon oxide film treated with hydrogen peroxide.

<Test Example>
I. Development of a nocturnal polyuria model

A nocturnal polyuria model was needed to study the pharmacological effects of silicon microparticles, but there had been only one report on an animal model of nocturnal polyuria (Ihara T. et al. Neurourol Urodyn. 2017 Apr;36(4):1034-1038). Because this model uses clock gene-modified mice, there is a possibility that circadian rhythms are abnormal, but sleep disorders are not thought to be closely related to nocturnal polyuria. Recognizing the need for a nocturnal polyuria model that mimics the pathology of human nocturnal polyuria, we established a new nocturnal polyuria model.

Human nocturnal urine volume is the volume of urine voided between the time a person goes to bed and the time they wake up (excluding the last urine voided before going to bed, but including the first urine voided after waking up). The mouse urine volume equivalent to human nocturnal urine volume is the volume voided during the resting period, which is daytime urine volume because mice are nocturnal. In this test example, the mouse's resting period urine volume was defined as the volume voided between 8:00 AM and 8:00 PM. The resting period urine volume ratio (the ratio of the amount of urine voided during the resting period to the daily urine volume) was calculated as an index for mice corresponding to the human nocturnal polyuria index. In this test example, mouse urine volume was calculated using the aVSOP method (Negoro H, et al. Nat Com, 2017; Takezawa K, et al. Am J Physiol Renal Physiol. 2014).

A. Salt Loading and Resting Period Urinary Volume Ratio in Mice High daily salt intake in urologists aged 40 years or older increases the nocturnal polyuria index (the ratio of nighttime urine volume to daily urine volume) (Matsuo T, et al. Int J Urol. 2017). However, even with the same salt intake, nocturnal polyuria does not occur in younger individuals, but only in elderly individuals. Therefore, the inventors examined whether the resting period urine volume ratio (resting period urine volume/daily urine volume) increases when elderly mice are fed a high-salt diet.

19-week-old and 80-week-old mice (C57BL/6J, male) were obtained from SLC (Japan SLC Co., Ltd.). They were fed a standard diet (MF, manufactured by Oriental Yeast Co., Ltd., containing 0.2% salt) or a 1% high-salt diet (same as the standard diet except for the salt content), and were allowed to eat and drink ad libitum. Urine volume was calculated after two weeks. The resting phase urine volume rate and daily urine volume are shown in Figure 4. Only the elderly mice (80-week-old mice) developed nocturnal polyuria due to salt loading.

B. Comparison of Urinary NOx in Aged and Young Mice Next, we examined the amount of urinary nitrogen oxides (NOx) ( ^NO2- , ^NO3- ). Urinary nitrogen oxides (NOx) ( ^NO2- , ^NO3- ) are an indicator of vascular aging (Toshio Hayashi, Journal of Geriatrics, 2011; 48: 142-145). Urinary NOx in aged mice was measured using the Griess method (Dojindo Laboratories, Griess Reagent Kit), and the daily excretion amount (creatinine-corrected value (μmol/mg Cr)) was calculated. Aged mice had significantly lower NOx levels than young mice (Figure 5). It was revealed that aged mice had a decrease in urinary NOx levels due to the progression of vascular aging.

C. Relationship between urinary NOx, salt intake, and nocturnal polyuria in humans We investigated the NOx concentration in human urine and the relationship between salt intake and nocturnal polyuria in a group with low urinary NOx concentration (a group with advanced vascular aging).

The amount of NOx in daily urine collected from 27 humans (hospitalized patients) (10 men, 17 women, mean age 61 years) with their consent was examined. Subjects were divided into a low NOx group (below the median (0.28 μmol/mgCr)) and a high NOx group (above the median), and the relationship with estimated salt intake was examined for each group. Estimated salt intake was calculated from the amount of Na excreted in the daily urine. A positive correlation was shown between salt intake and the nocturnal polyuria index in the low NOx group (a group with advanced vascular aging), but no correlation was shown in the high NOx group (Figure 6). It has been revealed that excessive salt intake can also cause nocturnal polyuria in humans in the low NOx group.

D. Establishment of an Animal Model of Nocturnal Polyuria We attempted to create a nocturnal polyuria model by administering a nitric oxide synthase inhibitor and excessive salt to mice.

19-week-old mice (C57BL/6J, male) were obtained from SLC. After being introduced into the experimental facility, they were fed either a 1% high-salt diet (described above in A) or a standard diet ad libitum. The high-salt diet mice were given the nitric oxide synthase inhibitor L-NAME (NG-nitro-L-arginine methyl ester hydrochloride) (Sigma-Aldrich Japan) dissolved in water at 0.5 mg/mL (ad libitum drinking water). Two weeks after the initiation of the high-salt diet, urine volume was measured and the resting period urine volume ratio was calculated. The results are shown in Figure 7. The resting period urine volume ratio was significantly higher in the nocturnal polyuria model group compared to the control group. All mice had a resting period urine volume ratio of 0.27 or higher, indicating resting period polyuria (nocturnal polyuria). A polyuria model mimicking human nocturnal polyuria was successfully established.

II. Preparation of silicon microparticle-containing food The silicon microparticles (silicon crystallites and their aggregates) produced in Example 2 were mixed into Oriental Yeast Co., Ltd., model number AIN93M, to a concentration of 2.5 wt%. Furthermore, citric acid aqueous solution (pH 4) was added in an amount of about 0.5 wt% relative to the total amount of the silicon microparticles and the feed, and kneaded using a known kneading device to obtain silicon microparticle-containing food. The silicon microparticles produced in Example 2 were mixed into the 1% high-salt food described in I above to a concentration of 1 wt%, and a silicon microparticle-containing high-salt food was obtained in the same manner as above.

III. Pharmacological Effects of Silicon Microparticles The effects of silicon microparticles were investigated using the nocturnal polyuria model prepared in I above. The nocturnal polyuria model was fed the 1% high-salt diet described in I above, or the high-salt diet containing silicon microparticles described in II above. Control mice were fed the standard diet described in I above, or the silicon microparticle-containing diet described in II above. Two weeks after the start of each diet, urine volume was measured using the aVSOP method, and the resting period urine volume rate of each group was compared (Figure 8). The resting period urine volume rate was significantly lower in the group administered with silicon microparticles to the nocturnal polyuria model compared to the nocturnal polyuria model group (Figure 9). These findings demonstrate that administration of silicon microparticles improves nocturnal polyuria.

IV. Mechanism of Action: NaCl cotransporter (NCC) and phosphorylated NCC (pNCC) were quantified by Western blotting. Kidneys were removed from the nocturnal polyuria model and the silicone microparticle-administered nocturnal polyuria model, and proteins were extracted. Proteins were then electrophoresed and transferred to a membrane. Protein bands were detected using an anti-NCC antibody (Millipore) and an anti-phosphorylated NCC antibody (Phospho Solution), and the ratio of phosphorylated NCC to NCC (pNCC/NCC) was calculated (Figure 10). Compared with the nocturnal polyuria model, the silicone microparticle-administered nocturnal polyuria model showed a significant decrease in phosphorylated NCC. This formulation is believed to improve nocturnal polyuria through a completely new mechanism distinct from the conventional drug desmopressin.

In countries with aging societies, the number of patients suffering from nocturia is extremely large, and it is expected that the preventive or therapeutic agent of the present invention will lead to improvements in the quality of life and survival rate of many elderly people.

Claims (12)

A preventive or therapeutic agent for nocturia caused by nocturnal polyuria, which contains silicon microparticles. The preventive or therapeutic agent according to claim 1 , which is used to prevent or treat nocturia by regulating nighttime urine volume. 3. The preventive or therapeutic agent according to claim 1 , wherein the silicon microparticles are microparticles containing elemental silicon capable of generating hydrogen upon contact with water. 4. The preventive or therapeutic agent according to claim 1, wherein the silicon microparticles have a silicon oxide film formed on the surface thereof. 5. The preventive or therapeutic agent according to claim 1, wherein the silicon microparticles are silicon microparticles and/or aggregates of the silicon microparticles. 6. The preventive or therapeutic agent according to claim 5, wherein the silicon microparticles are microparticles made of simple silicon with a silicon oxide film formed on the surface thereof. 5. The preventive or therapeutic agent according to claim 1, wherein the silicon microparticles are porous silicon particles. The preventive or therapeutic agent according to any one of claims 1 to 7, wherein the silicon microparticles are hydrophilically treated silicon microparticles. The preventive or therapeutic agent according to any one of claims 1 to 8, which is for oral administration. A pharmaceutical composition for preventing or treating nocturia caused by nocturnal polyuria, comprising silicon microparticles. A medical device containing silicon microparticles for the prevention or treatment of nocturia caused by nocturnal polyuria . A food or beverage containing silicon microparticles for preventing or treating nocturia caused by nocturnal polyuria .