Sperm motility improver

Abstract

To provide a motility improver for male sperms.SOLUTION: The male sperm motility enhancing agent of the present invention contains nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) as essential components. The agent for improving sperm motility can be used for infertility treatment of an adult male with reduced sperm motility and / or reduced sperm concentration, or for pregnancy activity (fertility) of an adult male. Another application is in the breeding of domesticated animals.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to an agent for improving sperm motility. [Background technology]

[0002] Japan is facing a declining birthrate and an aging population, and the population decline is expected to continue. Countermeasures are attracting attention, with infertility treatment becoming eligible for insurance coverage in 2022. In recent years, understanding of male sperm has advanced, and infertility treatment is now required to address male infertility. Semen analysis has revealed that the number of men experiencing infertility due to decreased sperm concentration, decreased sperm motility, and an increased rate of abnormal sperm is on the rise.

[0003] Thus, there is an urgent need to establish new methods for improving sperm function, but there are few examples of sperm research, and the reality is that we are currently exploring the establishment of effective methods for improvement. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2016-37473 [Patent Document 2] Japanese Patent Application Publication No. 2018-172346 [Non-patent literature]

[0005] [Non-Patent Document 1] Domestic Journal Author (Hamada Takeyuki) Title of paper: "The effect of 5-aminolevulinic acid (ALA) on improving semen findings" Publication name (Journal of the Japanese Society for Reproductive Medicine) Publication year (2017) Vol.62, No.4 p.208, Abstract No.O-132 Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a useful sperm motility enhancer that can improve sperm motility. [Means for solving the problem]

[0007] The gist of the present invention is as follows. (1) The sperm motility enhancer of the present invention is characterized by being a combination drug containing nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) as essential ingredients. (2) When the total amount of the three components, nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA), is taken as 1, the three components are contained in a weight ratio of 0.25 to 0.8:0.06 to 0.8:0.005 to 0.80. (3) Any of the nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) may be contained in the form of a derivative or salt. (4) The sperm motility enhancer is used for infertility treatment in adult men with reduced sperm motility and / or reduced sperm concentration, or for fertility treatment in adult men. (5) The sperm motility enhancer is used for breeding captive animals. (6) The sperm motility enhancer is added to semen. (7) The sperm motility enhancer is in the form of a tablet, powder, granule, dry syrup, capsule, liquid, or gel, and is administered orally or parenterally. [Effects of the Invention]

[0008] According to the present invention, a useful sperm motility enhancer can be provided by preparing a combination containing nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) as essential ingredients. As is clear from the results of the examples described below, it has been confirmed that the combination of the three ingredients of the present invention is an optimal combination that is superior in improving sperm motility compared to nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) individually. Moreover, each of these ingredients is safe for the human body and can be used with confidence. From the above, the present invention is effective as an agent for improving sperm function for, for example, infertile men and men trying to conceive (try to conceive), and is also effective for, for example, breeding animals.

[0009] As is clear from the results of the examples described below, the sperm motility enhancer of the present invention is not only excellent in improving sperm motility, but has also been confirmed to be superior in increasing ATP (Adenosine tri-phosphate) production and suppressing ROS (Reactive Oxygen Species) production compared to the individual use of nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA). [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 is an explanatory diagram regarding the action of a sperm motility enhancer according to an embodiment. [Figure 2] 1 is a graph showing the results of Test Example 1 confirming the effect of a sperm motility enhancer. [Figure 3] 1 is a graph showing the results of Test Example 2 confirming the effect of a sperm motility enhancer. DETAILED DESCRIPTION OF THE INVENTION

[0011] The "sperm motility enhancer" according to this embodiment will be described in detail below. However, the technical scope of the present invention is not limited to the following embodiments.

[0012] The sperm motility enhancer according to this embodiment (hereinafter referred to as "sperm motility enhancer") is a combination drug containing three essential ingredients: nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA).

[0013] Nicotinamide mononucleotide (NMN) is, for example, represented by the following chemical formula: [ka] Coenzyme Q10 is, for example, represented by the following chemical formula: [ka] 5-aminolevulinic acid (5-ALA) is, for example, represented by the following chemical formula: [ka]

[0014] The relative ratios of nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA), when the total amount of the three components is taken as 1, are preferably set at a mass ratio of 0.25-0.80:0.06-0.80:0.005-0.80, for example. A ratio of 1:1:1 is more preferred. Nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) are preferably contained in a form that can be represented by the above-mentioned chemical formula, but this is not limited thereto. For example, at least one component or all three components may be contained in the form of a derivative (i.e., a compound modified to a degree that does not significantly alter the parent structure or properties) or salt. Salts of the derivatives may also be used.

[0015] The sperm motility enhancer as a combination is preferably formulated by mixing or the like so that the total amount of the three components (including derivatives or salts) of nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) is 0.001% by mass to 100% by mass, preferably in the above ratio. Other components include compounds added as other functional ingredients, additives used in tableting, solvents used in liquid formulations, and additives used to maintain stable storage conditions for the product.

[0016] Nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) are already used as ingredients in supplements and the like, although their purposes and uses are different from those of the present embodiment, and are known to be safe for the human body when taken orally or parenterally (for example, by applying to the skin). The present embodiment is novel in that it combines these three components from known compounds, and it has been found that combining them into a single compound provides a greater effect on improving sperm motility than administering each component individually (see the results of the Examples below for details).

[0017] Sperm motility enhancers can be used in infertility treatment for adult men or in pregnancy efforts (trying to conceive) for adult men. They are particularly useful for infertility treatment and pregnancy efforts for adult men with reduced sperm motility and / or reduced sperm concentration. Sperm motility enhancers can be used in artificial insemination in general infertility treatment. As a specific example, semen obtained from a man is washed, concentrated, etc., and then injected into the woman's uterus together with a culture medium and a sperm motility enhancer. This has the advantage that it is only necessary to add a sperm motility enhancer to general artificial insemination. It can also be used in in vitro fertilization as assisted reproductive technology, not just general infertility treatment.

[0018] The form of use of the sperm motility enhancer is not limited to the above-mentioned artificial insemination, etc., and it may be used as an oral or parenteral agent. Specific examples include tablets, powders, granules, dry syrup, capsules, liquid agents, and gel agents. When used as a parenteral agent, it may be made into a gel and applied to the male genitals during sexual intercourse, but is not limited thereto.

[0019] Here, the sperm motility improving effect of the sperm motility improver of this embodiment will be described with reference to Figure 1. Figure 1 shows the effects of each component, nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA), and also shows the organic synergistic effect of each component when the three components are combined. This mechanism is inferred from the results of the Examples described below. As shown in Figure 1, we hypothesize that the organic synergistic effects of the three-component combination of nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) contribute to mitochondrial activity. More specifically, nicotinamide mononucleotide (NMN) works with vitamin B3 to produce NAD, which captures hydrogen ions in the TCA cycle. Coenzyme Q10 moves freely within mitochondria and facilitates the transfer of electrons between complexes. Cytochrome C, which receives electrons from coenzyme Q10, is a type of heme protein containing heme iron. 5-aminolevulinic acid (5-ALA) is the source of this heme iron. NAD, coenzyme Q10, and cytochrome C (heme iron) contribute to ATP production and are important components for activating mitochondria. The present inventors have conducted extensive research into whether mitochondrial activation can also be applied to improving sperm motility, and as a result have discovered a new use for the sperm motility improver of this embodiment and confirmed its effectiveness.

[0020] As described above, according to this embodiment, an effective sperm motility improver can be provided by using a combination containing nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) as essential ingredients. That is, it is effective as a sperm function improver for, for example, infertile men and men trying to conceive. Furthermore, it is also effective for, for example, breeding captive animals. The dosage form for animals may be the same as that described above. [Example]

[0021] Next, the results of the test to confirm the effect of the sperm motility enhancer according to this embodiment will be described. In this example, sperm from asthenozoospermia patients were used with the approval of the Tokushima University Hospital Life Science and Medical Research Ethics Committee. Analysis was performed using the Sperm Motility Analysis System (SMAS), which allows for objective numerical analysis.

[0022] Preparation of sperm motility enhancer As the sperm motility enhancer of Example 1, a combination preparation (P3) containing three components, nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA), was prepared. For the efficacy confirmation test, the concentrations of the three components were adjusted to 0.001% by mass, 0.01% by mass, and 0.1% by mass, respectively. The concentrations were adjusted using m-HTF Medium for sperm (Nihon Medical Equipment, product number: 9600-002).

[0023] Measurement of ATP levels in sperm ATP production was analyzed using the ATP Assay Kit-Luminescence (Dojindo Laboratories, Part Number: A550). 100 μL of the ATP standard solution and each sperm suspension was added to wells of a 96-well white microplate. ATP standard solutions were prepared by serial dilution in Modified HTF Medium to 2.5, 1.25, 0.625, 0.313, 0.156, 0.078, 0.039, and 0 μM. Next, 100 μL of working solution was added to each well. The microplate was covered with aluminum foil, shaken for 2 minutes, and then incubated at room temperature for 10 minutes. Luminescence was measured using a SpectraMAX multimode plate reader. TM Measurement was performed using i3 (Molecular Devices Japan). Cellular ATP concentration was calculated from the calibration curve.

[0024] Measurement of sperm mitochondrial ROS levels MitoSOX TM Mitochondrial Superoxide Indicators for live-cell imaging (M36008, Thermo Fisher Scientific KK) were used to analyze the intracellular production of sperm mitochondrial ROS. Sperm samples were immersed in 500 μL of MitoSOX. TM The cells were incubated with Red reagent working solution (5 μM) at 37°C for 10 minutes in the dark. TM Mitochondrial ROS levels were analyzed by flow cytometry using a 486 / 42 nm bandwidth filter on a Becton Dickinson flow cytometer. A total of 10,000 sperm-specific events were analyzed, and the positive rate was calculated.

[0025] As Comparative Examples 1 to 3 for the sperm motility enhancer of Example 1, similar tests were carried out on nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) individually. These are referred to as Comparative Example 1: NMN, Comparative Example 2: Q10, and Comparative Example 3: 5-ALA.

[0026] -Effects on sperm motility The results of the motility rate at each concentration (0% by mass (untreated), 0.001% by mass, 0.01% by mass, 0.1% by mass, 0.1% by mass) of Example 1 (P3), Comparative Example 1 (NMN), Comparative Example 2 (Q10), and Comparative Example 3 (5-ALA) are shown in Figure 2. As shown in the results of Figure 2, the motility rate decreased in a time-dependent manner when untreated. In each example, the motility rate was maintained 1 hour and 3 hours after mixing of each agent. Among them, Example 1 (P3) showed an increase in motility rate (particularly notable at 0.01% by mass). This effect of increasing motility is exceptional and not seen in Comparative Examples 1 to 3.

[0027] -Effects on ATP production and ROS production Figure 3 shows the results of measuring ATP production and ROS production at 0.01% by mass, at which the effect on sperm motility was evident, and one hour after mixing with the drug. As can be seen from the results in Figure 3, ATP production was unchanged in the cases of Comparative Examples 1 to 3 when used alone compared to the untreated group. On the other hand, APT production was significantly increased in the combination drug of Example 1 (P3) compared to the cases of Comparative Examples 1 to 3 when used alone. Furthermore, with regard to ROS production, although ROS production was suppressed in Comparative Examples 1 to 3 compared to the untreated group, it was clear that the combination drug of Example 1 (P3) had a higher suppression rate.

Claims

1. A sperm motility enhancer characterized by being a combination drug containing nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA) as essential ingredients.

2. The sperm motility improver according to claim 1, characterized in that the three components, nicotinamide mononucleotide (NMN), coenzyme Q10, and 5-aminolevulinic acid (5-ALA), are contained in a mass ratio of 0.25 to 0.80:0.06 to 0.80:0.005 to 0.80, where the total amount of the three components is 1.

3. The sperm motility enhancer according to claim 1, wherein any one of the nicotinamide mononucleotide (NMN), the coenzyme Q10, and the 5-aminolevulinic acid (5-ALA) may be contained in the form of a derivative or a salt.

4. The sperm motility improver according to any one of claims 1 to 3, characterized in that it is used for infertility treatment in adult men with reduced sperm motility and / or reduced sperm concentration, or for pregnancy efforts in adult men.

5. The sperm motility improver according to any one of claims 1 to 3, characterized in that the sperm motility improver is used for breeding animals kept in captivity.

6. The sperm motility improver according to any one of claims 1 to 3, characterized in that the sperm motility improver is used by adding it to semen.

7. The sperm motility enhancer according to any one of claims 1 to 3, characterized in that the sperm motility enhancer is in the form of a tablet, powder, granule, dry syrup, capsule, liquid, or gel, and is administered orally or parenterally.

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