Promoter or improver for decidualization of endometrium

Abstract

The invention provides a preparation capable of promoting normal deciduation of endometrium. An agent for promoting or ameliorating endometrial deciduation, which contains an effective amount of pyrroloquinoline quinone or a salt thereof, is prepared. In addition, a cell fusion promoting or ameliorating agent containing an effective amount of pyrroloquinoline quinone or a salt thereof is prepared.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480075517.9 (22) Application Date 2024.11.29 (30) Priority Data 2023-202970 2023.11.30 JP (85) PCT International Application Entering National Phase Date 2026.05.28 (86) PCT International Application Application Data PCT / JP2024 / 042312 2024.11.29 (87) PCT International Application Publication Data WO2025 / 116006 JA 2025.06.05 (71) Applicant: National University Corporation, Hiroshima University, Japan; Applicant School Corporation: Tokyo University of Pharmacy and Pharmaceutical Sciences, Japan; Rohto Pharmaceutical Co., Ltd. (72) Inventors: Masahiro Shimada, Kazuya Kusama (74) Patent Agency: Beijing J&J Intellectual Property Agency Co., Ltd., 11227; Patent Attorney: Qing Zhao (51) Int.Cl. A61K 31 / 475 (2006.01) A61P 15 / 00 (2006.01) A61P 15 / 08 (2006.01) (54) Invention Title: Endometrial Decidualization Promoter or Improver (57) Abstract: This invention provides an agent capable of promoting normal decidualization of the endometrium. An endometrial decidualization promoter or improver containing an effective amount of pyrroloquinoline quinone or its salt is prepared. In addition, a cell fusion promoter or improver containing an effective amount of pyrroloquinoline quinone or its salt is also prepared. Claims 1 page, Description 13 pages, Drawings 5 ​​pages, CN 122295102 A 2026.06.26 CN 1 22 29 51 02 A 1. A promoter or improver of endometrial decidualization, containing an effective amount of pyrroloquinoline quinone or a salt thereof. 2. An anti-aging agent for the endometrium, containing an effective amount of pyrroloquinoline quinone or a salt thereof. 3. The agent according to claim 2, wherein the anti-aging of the endometrium is by inhibiting the transformation of endometrial cells into senescent cells. 4. The agent according to claim 2, wherein the anti-aging of the endometrium is by clearing senescent cells from the endometrium. 5. A promoter or improver of placental trophoblast cell fusion, containing an effective amount of pyrroloquinoline quinone or a salt thereof. 6. A fertility enhancer, containing an effective amount of pyrroloquinoline quinone or a salt thereof, which enhances fertility by promoting or improving endometrial decidualization and / or promoting or improving placental trophoblast cell fusion. 7. The agent according to claim 6, wherein the improvement in fertility is defined as improved implantation or continued implantation. 8. The agent according to any one of claims 1 to 7, wherein it is ingested for at least 7 to 21 days after the first day of menstruation.9. The agent according to any one of claims 1 to 7, which is ingested for at least 14 to 35 days after the first day of menstruation. Claims 1 / 1 page 2 CN 122295102 A Endometrial decidualization promoter or improver Technical Field

[0001] The present invention relates to an endometrial decidualization promoter or improver. Background Art

[0002] According to statistics from the Japanese Society of Obstetrics and Gynecology, the number of infertility treatment cases per year has increased sharply from about 10,000 in 2004 to more than 400,000 in 2015, and has remained at a high level since then, at about 450,000, indicating that a large number of couples are suffering from infertility problems.

[0003] In Japan, since 2022, infertility treatments such as artificial insemination ("general infertility treatment") and in vitro fertilization (IVF / CI) ("assisted reproductive technology") have been included in insurance coverage, aiming to alleviate the economic burden of many infertility problems.

[0004] However, infertility treatments covered by insurance for women are subject to age and frequency restrictions. Therefore, even if treatment is covered by insurance, it cannot be said that women's concerns about infertility have been fully resolved.

[0005] Female infertility is classified into ovarian factors, fallopian tube factors, and uterine factors according to its etiology. However, the ovary, fallopian tube, and uterus influence each other. Without elucidating their mechanisms, it is difficult to elucidate the complex physiological phenomenon of ovulation-fertilization-implantation-embryonic development, thus making it difficult to fundamentally solve infertility.

[0006] In this complex physiological phenomenon, the inventors focused on the decidualization of endometrial cells, believing that if normal decidualization cannot occur, it may lead to implantation failure or miscarriage. They established an in vitro experimental system to simulate the decidualization of human endometrial cells and studied "normal decidualization" (Non-Patent Literature 1).

[0007] On the other hand, pyrroloquinoline quinone (PQQ) is an organic molecule discovered in bacteria as a redox coenzyme. It is a highly versatile and safe food ingredient. Based on existing research on PQQ and infertility, it is known to improve sperm motility (Patent Document 1) and enhance the normality of eggs or ovaries (Patent Document 2).

[0008] Prior Art Documents

[0009] Patent Documents

[0010] Patent Document 1: International Publication No. 2020 / 013086

[0011] Patent Document 2: International Publication No. 2020 / 262325

[0012] Non-Patent Documents

[0013] Non-Patent Document 1: Kazuya Kusama et al. Biochemical and Biophysical Research Communications 571 (2021) 174-180 Summary of the Invention

[0014] According to the above patent documents, it is shown that by inhibiting DNA damage caused by oxidation of sperm or egg through the antioxidant effect of PQQ, sperm with high motility or eggs with high fertilization capacity can be obtained.

[0015] However, for example, even if normal sperm and eggs are successfully fertilized, if the aforementioned "normal decidualization of the endometrium" does not occur, the implantation period (an extremely precise timing) will be missed, resulting in failure to implant. Furthermore, if "normal decidualization of the endometrium" does not occur, excessive decidualization may lead to endometrial cell senescence. That is, promoting endometrial decidualization while inhibiting abnormal cell senescence as much as possible is "promoting normal decidualization of the endometrium," which is a necessary condition for implantation. Specification 1 / 13 pages 3 CN 122295102 A

[0016] Therefore, the inventors conducted in-depth research on whether PQQ can "promote normal decidualization of the endometrium," and found that PQQ can significantly promote decidualization while inhibiting the expression of metabolic waste and senescence genes, which are indicators of cell senescence, and found that PQQ promotes implantation by "promoting normal decidualization of the endometrium." In addition, the inventors found that PQQ can promote the fusion of placental trophoblast cells, which are crucial for the completion of pregnancy. Then, focusing on the connection between PQQ and improved fertility, the present invention was completed.

[0017] That is, the preferred embodiment of the present invention provides the following oral composition.

[0018] [1] An agent for promoting or improving decidualization of the endometrium, containing an effective amount of pyrroloquinoline quinone or a salt thereof.

[0019] [2] An anti-aging agent for the endometrium, containing an effective amount of pyrroloquinoline quinone or a salt thereof.

[0020] [3] The agent according to [2], wherein the anti-aging of the endometrium is to inhibit the transformation of endometrial cells into senescent cells.

[0021] [4] The agent according to [2], wherein the anti-aging of the endometrium is to remove senescent cells from the endometrium.

[0022] [5] An agent for promoting or improving the fusion of placental trophoblast cells, containing an effective amount of pyrroloquinoline quinone or a salt thereof.

[0023] [6] A fertility enhancer containing an effective amount of pyrroloquinoline quinone or a salt thereof, which enhances fertility by promoting or improving decidualization of the endometrium and / or promoting or improving placental trophoblast cell fusion.

[0024] [7] The agent according to [6], wherein the enhancement of fertility is implantation improvement or persistence of implantation status.

[0025] [8] The agent according to any one of [1] to [7] is taken for use during a period of at least 7 to 21 days after the first day of menstruation.

[0026] [9] The agent according to any one of [1] to [7] is taken for use during a period of at least 14 to 35 days after the first day of menstruation.

[0027] According to the present invention, since PQQ can significantly promote decidualization while also inhibiting the expression of metabolic waste and senescence genes, which are indicators of cellular senescence, an agent that promotes or improves decidualization of the endometrium can be provided. In addition, an agent that promotes or improves placental trophoblast cell fusion can be provided.

[0028] Figure 1 shows the cytotoxicity test results in Example 1.

[0029] Figure 2 shows the decidualization promotion test results of primary endometrial stromal cells (pESCs) in Example 2-1.

[0030] Figure 3 shows the epithelial differentiation promotion test results of Ishikawa cells and EM1 cells in Example 2-2.

[0031] Figure 4 shows the inhibition test results of aging-related β-galactosidase associated with pESC decidualization in Example 3-1.

[0032] Figure 5 shows the inhibition test results of the aging gene p53 associated with pESC decidualization and the promotion test results of anti-aging genes in Example 3-2.

[0033] Figure 6 shows the gene expression test results in the endometrial cells of mice that drank PQQ in Example 4-1.

[0034] Figure 7 shows photographs and charts of immunostaining in the endometrium of mice that drank PQQ in Example 4-2.

[0035] Figure 8 shows the implantation rate evaluation results of mice drinking PQQ in Experiment 5.

[0036] Figure 9 shows the gene expression test results as an indicator of cell fusion in Experiment 6. Specification 2 / 13 pages 4 CN 122295102 A

[0037] Figure 10 is a photograph showing the cell fusion state of Experiment 7-1.

[0038] Figure 11 is a graph showing the change in the number of fused cells in Experiment 7-2. Detailed Description

[0039] [Promoter or Improver of Endometrial Decidualization]

[0040] The formulation of this method contains an effective amount of pyrroloquinoline quinone or its salt.

[0041] (Pyrroloquinoline quinone or its salt)

[0042] Pyrroloquinoline quinone is a known compound represented by the following Formula 1.

[0043]

[0044] Examples of salts of pyrroloquinoline quinone include alkali metal salts, alkaline earth metal salts, and ammonium salts.Examples of alkali metal salts include sodium, potassium, and lithium salts. Examples of alkaline earth metal salts include calcium and magnesium salts.

[0045] Alkali metal salts of pyrroloquinoline quinone or salts thereof are preferred, sodium salts of pyrroloquinoline quinone are more preferred, and disodium salts of pyrroloquinoline quinone are even more preferred.

[0046] Commercially available products of pyrroloquinoline quinone or salts thereof may also be used. In addition, one type of pyrroloquinoline quinone or salt thereof may be used alone, or two or more types may be used in combination.

[0047] From the viewpoint of significantly exerting the effects of the present invention, the content of pyrroloquinoline quinone or salt thereof relative to the total amount of the formulation may be, for example, 0.001% by mass or more, 0.005% by mass or more, 0.01% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.5% by mass or more, 1% by mass or more, etc.

[0048] In addition, the content of pyrroloquinoline quinone or its salt relative to the total amount of the formulation can be, for example, 30% or less by mass, 25% or less by mass, 20% or less by mass, 15% or less by mass, 10% or less by mass, 5% or less by mass, 1% or less by mass, 0.5% or less by mass, 0.2% or less by mass, etc.

[0049] In addition, the content of pyrroloquinoline quinone or its salt relative to the total amount of the formulation can be, for example, 0.001–30% by mass, 0.001–25% by mass, 0.001–20% by mass, 0.001–15% by mass, 0.001–10% by mass, 0.001–5% by mass, 0.001–1% by mass, 0.001–0.5% by mass, 0.001–0.2% by mass, 0.005–30% by mass, 0.005–25% by mass, 0.005–20% by mass, 0.005–15% by mass, 0.005–10% by mass, 0.005–5% by mass, 0.005–1% by mass, 0.005–0.5% by mass, 0.005–0.2% by mass, 0.01–30% by mass, 0.01–25% by mass, and 0.01–20% by mass. 0.01–15% by mass, 0.01–10% by mass, 0.01–5% by mass, 0.01–1% by mass, 0.01–0.5% by mass, 0.01–0.2% by mass, 0.05–30% by mass, 0.05–25% by mass, 0.05–20% by mass, 0.05–15% by mass, 0.05–10% by mass, 0.05–5% by mass, 0.05–1% by mass, 0.05–0.5% by mass, 0.05–0.2% by mass, etc.

[0050] The daily intake of pyrroloquinoline quinone or its salts may vary depending on the individual's condition (weight, age, sex, etc.), formulation, etc., but from the viewpoint of significantly maximizing the effects of the present invention, for example, amounts of 1 mg or more, 2 mg or more, 3 mg or more, 4 mg or more, 5 mg or more, 6 mg or more, 7 mg or more, 8 mg or more, 9 mg or more, 10 mg or more, 15 mg or more, 20 mg or more, etc.

[0051] In addition, the daily intake of pyrroloquinoline quinone or its salts may, for example, be less than 100 mg, less than 80 mg, less than 60 mg, less than 50 mg, less than 40 mg, less than 30 mg, less than 25 mg, etc. Instructions for Use, page 3 / 13, CN 122295102 A

[0052] In addition, the daily intake of pyrroloquinoline quinone or its salts may include, for example, 5-100 mg, 5-80 mg, 5-60 mg, 5-50 mg, 5-40 mg, 5-30 mg, 5-25 mg, 10-100 mg, 10-80 mg, 10-60 mg, 10-50 mg, 10-40 mg, 10-30 mg, 10-25 mg, 15-100 mg, 15-80 mg, 15-60 mg, 15-50 mg, 15-40 mg, 15-30 mg, 15-25 mg, 20-100 mg, 20-80 mg, 20-60 mg, 20-50 mg, 20-40 mg, 20-30 mg, 20-25 mg, etc.

[0053] As long as the effects of the present invention are not compromised, the formulation of this embodiment may contain pharmacologically active ingredients or physiologically active ingredients other than pyrroloquinoline quinone or its salts.

[0054] Specific examples of such pharmacologically or physiologically active ingredients include, for example, ubiquinone (coenzyme Q10), vitamin B1, vitamin B2, vitamin B6, folic acid, vitamin B12, vitamin C, vitamin D, vitamin A, vitamin E, niacin, calcium pantothenate, taurine, carnosine, anserine, balenine, citrulline, γ-aminobutyric acid, valine, leucine, isoleucine, glycine, arginine, ornithine, carnitine, glutamic acid, glutamine, creatine, carnitine, luteolin, quercetin, genistein, anthocyanins, resveratrol, diosgenin, isoflavone aglycones, isoflavones, lipoic acid, zinc, iron, calcium, selenium, astaxanthin, zeaxanthin, β-carotene, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, Linolenic acid, arachidonic acid, placental extract, pycnogenol (proanthocyanidins), or estrol, etc.As a pharmacologically active ingredient or physiologically active ingredient, from the viewpoint of further improving the effect of the present invention, it is preferably selected from one or more of ubiquinone (coenzyme Q10), vitamin B12, vitamin C, vitamin E, astaxanthin, zinc, carnitine, resveratrol, isoflavones, equol, pycnogenol (proanthocyanidins), vitamin D, iron, and folic acid; more preferably selected from one or more of ubiquinone (coenzyme Q10), astaxanthin, zinc, resveratrol, isoflavones, equol, pycnogenol (proanthocyanidins), vitamin D, iron, and folic acid; and even more preferably selected from one or more of zinc, vitamin D, iron, and folic acid. A single pharmacologically active ingredient or physiologically active ingredient may be used alone, or two or more may be used in combination.

[0055] [Uses]

[0056] The endometrium is a tissue necessary for the establishment and maintenance of pregnancy. During the menstrual cycle, it proliferates and thickens in response to ovarian steroid hormones, thereby changing into a state called the exfoliative membrane, which allows for implantation of a fertilized egg. During the proliferative phase (from the first day of menstruation to about 14 days after menstruation), the endometrium proliferates and thickens in preparation for the next ovulation; during the secretory phase (about 14 days after the first day of menstruation), the endometrium matures and awaits fertilization; during menstruation, the endometrium sheds and is expelled from the body. The endometrium is mainly composed of stromal cells and glandular epithelial cells, and the formation of a shedding membrane through the proliferation of stromal cells is called decidualization.

[0057] As mentioned above, even if normal sperm and egg successfully fertilize, if "normal decidualization of the endometrium" does not occur, the implantation period (an extremely precise timing) will be missed, resulting in implantation failure. In addition, if "normal decidualization of the endometrium" does not occur, excessive decidualization may lead to endometrial cell senescence. That is, promoting endometrial decidualization while inhibiting abnormal cell senescence as much as possible is "promoting normal decidualization of the endometrium," which is a necessary condition for implantation.

[0058] In later embodiments, primary endometrial stromal cells (pESCs) that maintained the phenotype of actual human endometrial stromal cells were used, and pyrroloquinoline quinone or its salt (hereinafter also referred to as PQQ) was found to be a component that could promote endometrial decidualization.

[0059] In Experiment 1, no toxicity of PQQ to pESCs was found. In addition, in Experiment 2-1, when maturation-stimulating components (Db: dibutyryl-cAMP, and / or MPA: medroxyprogesterone acetate) simulating stimulation by ovarian steroid hormones were applied to pESCs together with PQQ, it was found that even the lowest concentration (1 nM) of PQQ significantly promoted decidualization of pESCs.

[0060] In the simulated in vitro decidualization test system of the later embodiments, although Db or MPA is used to simulate the stimulation brought about by ovarian steroid hormones, in vivo, the stimulation of progesterone begins to rise from the late proliferative phase and rises sharply in the secretory phase (approximately 14 days after the first day of menstruation). Therefore, based on the results of the later embodiments, PQQ is preferably taken from at least 7 days after the first day of menstruation, and preferably continued to be taken after 14 days from the first day of menstruation. In addition, in another embodiment, it is preferable to take it until the period when the normal decidualization of the endometrium is completed and the endometrium matures, i.e., 21 days after the first day of menstruation, but it is also possible to take it up to 35 days, and it is not limited thereto.

[0061] In addition, in the later embodiments, when the maturation stimulating components (Db and / or MPA) were applied to pESC together with PQQ, aging-related β-galactosidase was significantly inhibited, thus confirming the inhibition of the aging gene p53 and the overactivity of the anti-aging gene SIRT1. Therefore, it is shown that PQQ has the function of inhibiting aging (anti-aging) during the maturation process of endometrial tissue. It is thus confirmed that PQQ has the effect of inhibiting endometrial cell aging and clearing senescent cells.

[0062] In addition, in the later embodiments, it was confirmed not only in vitro but also in animal models that PQQ promotes epithelial differentiation of the endometrium and assists and promotes the implantation of the endometrium. It is strongly speculated that this is because oral intake of PQQ promotes normal decidualization of the endometrium, inhibits cell aging, and brings about a better maturation to implantation period, thereby improving fertility, improving implantation, and maintaining implantation status.

[0063] (Additives, etc.)

[0064] In addition to the above-mentioned components, the formulation of this embodiment may also contain various additives as long as it does not impair the effects of the present invention.

[0065] Specific examples of such additives include, for instance, excipients, binders, disintegrants, lubricants, colorants, flavoring agents, odorants, sugars, sugar alcohols / polyols, high-sweetness sweeteners, oils, emulsifiers, thickeners, acidulants, fruit juices, etc.

[0066] Examples of excipients include lactose, sucrose, sodium chloride, glucose, starch, gelatin, calcium carbonate, kaolin, crystalline cellulose, silicic acid, etc.

[0067] Examples of binders include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, cellulose, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, calcium phosphate, polyvinylpyrrolidone, etc.

[0068] Examples of disintegrants include dry starch, sodium alginate, agar powder, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, glyceryl monostearate, lactose, etc.

[0069] Examples of lubricants include talc, stearates, borax, and polyethylene glycol.

[0070] Examples of flavoring agents include sucrose, orange peel, citric acid, and tartaric acid.

[0071] Examples of sugars include sucrose, isomerized sugars, glucose, fructose, palaginose, trehalose, lactose, and xylose.

[0072] Examples of sugar alcohols / polyols include sorbitol, xylitol, erythritol, lactitol, palaginitol, reduced maltose syrup, reduced maltose syrup, glycerol, and propylene glycol.

[0073] Examples of high-sweetness sweeteners include aspartame, stevia, acesulfame potassium, and sucralose.

[0074] Examples of oils include safflower oil, grapeseed oil, sunflower seed oil, olive oil, corn oil, sesame oil, soybean oil, rapeseed oil, and perilla oil.

[0075] Examples of emulsifiers include sucrose fatty acid esters, glycerol fatty acid esters, and lecithin.

[0076] Examples of thickeners include carrageenan, xanthan gum, guar gum, pectin, and locust bean gum.

[0077] Examples of acidulants include citric acid, lactic acid, and malic acid. Examples of fruit juices include lemon juice, orange juice, and berry juice.

[0078] One additive may be used alone, or two or more may be used in combination.

[0079] From the viewpoint of the stability of pyrroloquinoline quinone or its salts during formulation, the additive preferably contains oils.

[0080] (Dosage Form, etc.)

[0081] The dosage form of the preparation in this embodiment is not particularly limited. Examples include: oral preparations such as tablets (including orally disintegrating tablets, chewable tablets, lozenges, etc.), granules, powders, soft capsules, hard capsules, lozenges, gels, or liquids (including suspensions, emulsions, syrups, etc.); external preparations such as ointments, suppositories, patches, and sprays; and injections. From the viewpoint of ease of handling and further improving the effect of the present invention, oral preparations are preferred, and tablets, granules, soft capsules, and hard capsules are more preferred.

[0082] The preparation in this embodiment can be used, for example, as an ingredient in pharmaceuticals, quasi-pharmaceuticals, cosmetics, and food and beverage products (beverages, foods). In addition, the preparation in this embodiment can also be used, for example, as pharmaceutical preparations, quasi-pharmaceutical preparations, specific health foods, nutritional functional foods, foods for the elderly, special-purpose foods, functionally labeled foods, health supplements (supplements), food preparations (e.g., compressed candy), and obvious foods. Furthermore, the pharmaceutical preparation of this embodiment can also be used, for example, as an animal medicine or feed additive.

[0083] When the preparation of this embodiment is used as food, the food may be a food containing pyrroloquinoline quinone or its salt, and other ingredients as needed, in combination with a general food.Examples of such foods include solid foods such as cookies, biscuits, snacks, jelly, gummies, chocolate, chewing gum, candy, and cheese; and liquid foods such as nutritional drinks, fruit juices, tea drinks, coffee drinks, and milk drinks.

[0084] The formulation of this embodiment can be used for objects that require normal decidualization of the endometrium (e.g., humans; non-human animals such as cattle, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets). From the viewpoint of further improving the effect of the present invention, humans are preferred as the object. In addition, from the viewpoint of further improving the effect of the present invention, cattle, pigs, and horses are preferred among non-human animals, and pigs are more preferred.

[0085] [Manufacturing Method]

[0086] The manufacturing method of the formulation of this embodiment is not particularly limited, and known methods can be used. Specifically, a method can be given in which the components are heated and stirred to dissolve or disperse them as needed, mixed, cooled and stirred, and then left to stand at room temperature, but it is not particularly limited to this method.

[0087] [Administration Method]

[0088] The formulation of this embodiment is not limited, and can be stored and / or sold in a known container at a dosage of more than 1 week, more than 10 days, more than 14 days, more than 20 days, or more than 30 days.

[0089] The formulation of this embodiment is not limited, and the administration period can be, for example, at least 3 days, or more than 5 days, more than 7 days, more than 10 days, more than 14 days, more than 20 days, more than 30 days, or more than 35 days.

[0090] [Method for Promoting or Improving Endometrial Decidualization, Method for Anti-Aging of the Endometrium, and Method for Improving Fertility Caused by Anti-Aging of the Endometrium]

[0091] Pyrroloquinoline quinone or its salts have the effect of significantly promoting decidualization while also inhibiting the expression of metabolic waste and aging genes, which are indicators of cellular aging. Therefore, as an embodiment of the present invention, a method for promoting or improving endometrial decidualization, an anti-aging method for the endometrium, and a method for improving fertility caused by promoting endometrial decidualization are provided. These methods include the following steps: ingesting a preparation containing pyrroloquinoline quinone or its salt. The endometrium can be human endometrium or endometrium from non-human animals such as cattle, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets. From the viewpoint of further improving the effect of the present invention, human endometrium is preferred. It should be noted that the type, content, and daily intake of pyrroloquinoline quinone or its salt in the preparations of these embodiments, the type, content, and daily intake of other components, and the form of the preparation are as described above. Specification 6 / 13 pages 8 CN 122295102 A

[0092] [Placement trophoblast cell fusion promoter or improver]

[0093] The preparation of this method contains an effective amount of pyrroloquinoline quinone or its salt.

[0094] The definition, content, daily intake, and other conditions of pyrroloquinoline quinone or its salts are based on the information described in [Promoters or Improvers of Endometrial Decidualization].

[0095] As long as the effects of the present invention are not impaired, the formulation of this embodiment may also contain pharmacologically active or physiologically active ingredients other than pyrroloquinoline quinone or its salts.

[0096] Specific examples of such pharmacologically active or physiologically active ingredients are based on the information described in [Promoters or Improvers of Endometrial Decidualization].

[0097] [Uses]

[0098] The implantation process used to establish pregnancy can be roughly divided into three stages: attachment, adhesion, and infiltration of the fertilized egg in the endometrium, and also involves complex processes such as maternal immune regulation. During the infiltration process, the trophoblast cells of the fertilized egg that have moved into the endometrium continue to proliferate, differentiate, and fuse, continuously forming syncytiotrophoblast cells (cell fusion). That is, after the formation of the fertilized egg, the fusion of trophoblast cells in the uterus is indispensable for the establishment and maintenance of normal implantation. Therefore, promoting or improving placental trophoblast cell fusion helps to assist in the establishment of fertilized egg implantation, thereby assisting in the completion of pregnancy.

[0099] In the following examples, pyrroloquinoline quinone or its salts were found to be components that can promote placental trophoblast cell fusion.

[0100] In Experiment 6, it was confirmed that PQQ contributes to the increase in the expression of syncytin 2 and PAPPA as indicators during the process of inducing placental trophoblast cells to fuse with syncytiotrophoblast cells (fused trophoblast cells).

[0101] In the simulated in vitro placental trophoblast cell fusion test system in the following examples, although forskolin was used to stimulate cell fusion, in vivo, cell fusion is a phenomenon that occurs near the completion of implantation. It is strongly speculated that by promoting cell fusion that occurs from the beginning to the completion of the implantation process, fertility is further improved, involving the establishment and continuation of the implantation state.

[0102] (Additives, etc.)

[0103] In addition to the above-mentioned ingredients, the formulation of this embodiment may contain various additives as long as it does not impair the effects of the present invention.

[0104] Specific examples of such additives and other conditions are based on the contents described in [Promoters or Improvers of Endometrial Decidualization].

[0105] The dosage form of the formulation of this embodiment is also based on the contents described in [Promoters or Improvers of Endometrial Decidualization].

[0106] The formulation of this embodiment can be used, for example, as an ingredient in pharmaceuticals, quasi-pharmaceuticals, cosmetics, and food and beverage products (beverages, foods). In addition, the formulation of this embodiment can also be used, for example, as a pharmaceutical preparation, a quasi-pharmaceutical preparation, a food for specific health purposes, a nutritional functional food, a food for the elderly, a food for special purposes, a functionally labeled food, a health supplement (supplement), a food preparation (e.g., a candy tablet), or a distinct food.Furthermore, the agent of this embodiment can also be used, for example, as an animal medicine or feed additive.

[0107] When the preparation of this embodiment is used as a food, the food can be a food in which pyrroloquinoline quinone or its salt, and other ingredients as needed, are added to a general food. Examples of such foods include solid foods such as cookies, biscuits, snacks, jellies, gummies, chocolates, chewing gum, candies, and cheeses; and liquid foods such as nutritional drinks, fruit juices, tea drinks, coffee drinks, and milk drinks.

[0108] The preparation of this embodiment can be suitable for use in the subject matter of the specification 7 / 13 page 9 CN 122295102 A (e.g., humans; non-human animals such as cattle, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, and ferrets) that benefit from promoting or improving the fusion of placental trophoblast cells. From the viewpoint of further improving the effect of the present invention, humans are preferred as the subject matter. In addition, from the viewpoint of further improving the effect of the present invention, cattle, pigs, and horses are preferred among non-human animals, and pigs are more preferred.

[0109] [Manufacturing Method]

[0110] The manufacturing method of the formulation of this embodiment is not particularly limited, and known methods can be used. Specifically, a method can be given as follows: heating and stirring each component to dissolve or disperse it as needed, mixing it, cooling and stirring it, and then letting it stand at room temperature, but it is not particularly limited to this.

[0111] [Administration Method]

[0112] The formulation of this embodiment is not limited, and can be stored and / or sold in known containers in dosages of 1 week or more, 10 days or more, 14 days or more, 20 days or more, 30 days or more, or 35 days or more.

[0113] The formulation of this embodiment is not limited, and the administration period can be, for example, at least 3 days or more, or 5 days or more, 7 days or more, 10 days or more, 14 days or more, 20 days or more, 30 days or more, or 35 days or more.

[0114] [Methods for promoting or improving placental trophoblast cell fusion, methods for improving fertility caused by promoting or improving placental trophoblast cell fusion]

[0115] Pyrroloquinoline quinone or its salts can significantly promote the transformation of undifferentiated placental trophoblast cells into fused trophoblast cells. This can be verified, for example, by detecting increased expression of Syncytin 2 or PAPPA, which are markers of placental trophoblast cell fusion. Therefore, as an embodiment of the present invention, methods for promoting or improving placental trophoblast cell fusion and methods for improving fertility caused by promoting placental trophoblast cell fusion are provided, these methods comprising the step of: taking up a preparation containing pyrroloquinoline quinone or its salts. Placental trophoblast cells can be human cells or cells of non-human animals such as cattle, pigs, horses, sheep, goats, dogs, rabbits, mice, rats, guinea pigs, gerbils, hamsters, ferrets, etc. From the viewpoint of further improving the effects of the present invention, human cells are preferred.It should be noted that the types, contents, and daily intake of pyrroloquinoline quinone or its salts in the formulations of these embodiments, the types, contents, and daily intake of other components, and the formulation forms are as described above.

[0116] The present invention includes the following methods.

[0117] [1] A method for promoting or improving decidualization of the endometrium, wherein an effective amount of pyrroloquinoline quinone or its salt is administered.

[0118] [2] A method for preventing endometrial aging, wherein an effective amount of pyrroloquinoline quinone or its salt is administered.

[0119] [3] According to the method of [2], wherein the anti-aging of the endometrium is to inhibit the transformation of endometrial cells into senescent cells.

[0120] [4] According to the method of [2], wherein the anti-aging of the endometrium is to remove senescent cells from the endometrium.

[0121] [5] A method for promoting or improving the fusion of placental trophoblast cells, wherein an effective amount of pyrroloquinoline quinone or its salt is administered.

[0122] [6] A method for improving fertility, comprising administering an effective amount of pyrroloquinoline quinone or a salt thereof to improve fertility by promoting or improving decidualization of the endometrium and / or promoting or improving fusion of placental trophoblast cells.

[0123] [7] The method according to [6], wherein the improvement in fertility is an improvement in implantation or a continuation of the implantation state.

[0124] [8] The method according to any one of [1] to [7], wherein the administration is performed for at least 7 to 21 days after the first day of menstruation.

[0125] [9] The method according to any one of [1] to [7], wherein the administration is performed for at least 14 to 35 days after the first day of menstruation.

[0126]

[10] The use of pyrroloquinoline quinone or a salt thereof in the manufacture of an agent for promoting or improving decidualization of the endometrium.

[0127]

[11] Use of pyrroloquinoline quinone or its salt in the manufacture of an anti-aging agent for the endometrium.

[0128]

[12] According to the use of

[11] , wherein the anti-aging of the endometrium is the inhibition of endometrial cells from becoming senescent cells.

[0129]

[13] According to the use of

[11] , wherein the anti-aging of the endometrium is the removal of senescent cells from the endometrium.

[0130]

[14] Use of pyrroloquinoline quinone or its salt in the manufacture of a promoter or improver of placental trophoblast cell fusion.

[0131]

[15] Use of pyrroloquinoline quinone or its salt in the manufacture of a fertility enhancer, thereby enhancing fertility by promoting or improving endometrial decidualization and / or promoting or improving placental trophoblast cell fusion.

[0132]

[16] According to the use of

[15] , wherein the enhancement of fertility is improved implantation or a sustained implantation state.

[0133]

[17] Use according to any one of

[10] to

[16] , wherein the agent is administered for at least 7 to 21 days after the first day of menstruation.

[0134]

[18] Use according to any one of

[10] to

[16] , wherein the agent is administered for at least 14 to 35 days after the first day of menstruation.

[0135] Examples

[0136] The present invention will be specifically described below by way of examples and test examples, but the present invention is not limited to the following examples and test examples. Unless otherwise specified, the units of the amounts of each component in the table are mass%.

[0137] [Removal of endometrium and isolation of human primary endometrial stromal cells (pESC)]

[0138] In order to simulate the maturation of endometrium in vitro, primary cultured cells that maintain the phenotype of actual human endometrial stromal cells were used.

[0139] Human endometrial tissue was collected from female patients with endometriosis (n=3) under 45 years of age and with regular menstrual cycles. The collected endometrial tissue was washed with a Ca²⁺ / Mg²⁺-free Hanks' balanced salt solution (HBSS, manufactured by Fujifilm and Kojun Pharmaceutical Co., Ltd.), cut into small pieces, and digested for 2 hours at 37°C with type I collagenase (2.5 mg / ml, manufactured by Sigma Aldrich Japan) and DNase I (25 mg / ml, manufactured by Nippon Gene Co., Ltd.) in the presence of PSN (100 mg / ml penicillin, 100 mg / ml streptomycin, and 200 mg / ml neomycin; manufactured by Thermo Fisher Scientific). Undigested tissue was removed by passing the digestate through a 250 mm pore size filter, followed by filtration through a 38 mm filter to separate epithelial cells from stromal cells. The cells obtained in this way were used as primary endometrial stromal cells (pESCs).

[0140] [Cultivation of endometrial cell populations (human primary endometrial stromal cells, human endometrial epithelial cell line, and human endometrial glandular epithelial cell line)]

[0141] The above-mentioned pECS, human endometrial epithelial cell line (Ishikawa 3-H-12, JCRB cell bank, hereinafter referred to as "Ishikawa cells"), and human endometrial glandular epithelial cell line (EM-E6 / E7 / TERT-1, hereinafter referred to as "EM1 cells") were resuspended in Dulbecco modified Eagle medium / F12 (DMEM / F12) (1:1) medium (manufactured by Fujifilm and Koh Genuine Chemicals Co., Ltd.) supplemented with 10% fetal bovine serum (FBS), antibiotics, and antifungal drugs. The culture was then inoculated into gelatin-coated culture dishes at 37°C in humidified air containing 5% CO2.

[0142] [Stimulation of maturation of endometrial cell populations]

[0143] In order to evaluate the maturation of endometrial cell populations (pESC, Ishikawa cells, EM1 cells) in vitro, the following maturation stimulating components were added to the above-mentioned cultured cells to promote the differentiation of the endometrium in preparation for implantation. For pESC, butyryl-cAMP (500 mM, manufactured by Tokyo Chemical Industry Co., Ltd., hereinafter referred to as "Db") and progestin (1 mM, manufactured by Sigma Aldrich Japan, medroxyprogesterone acetate, hereinafter referred to as "MPA") were added to the culture medium on page 9 / 13 of the instruction manual, CN 122295102 A. For Ishikawa cells and EM1 cells, butyryl-cAMP (500 mM) was added to the culture medium, and various concentrations of PQQ were also added, and the cells were cultured for 48 hours.

[0144] [Cytotoxicity assay of pESC]

[0145] To evaluate the cytotoxicity of PQQ to pESC, the cell proliferation assay kit (Cell Counting Kit-8, manufactured by Dojin Chemical Research Institute Co., Ltd.) and the cytotoxicity assay kit (Cytotoxicity LDH Assay Kit-WST, manufactured by Dojin Chemical Research Institute Co., Ltd.) were used in the cultured cells described above, and the assays were performed according to the manufacturer's instructions.

[0146] [Staining of senescence-related β-galactosidase 1 in pESC]

[0147] To evaluate the degree of senescence of pESC, the cultured pESCs were fixed with 4% paraformaldehyde for 10 minutes and then washed twice with PBS. Staining was performed overnight at 37°C in phosphate-buffered saline adjusted to pH 6.0 with 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 1 mM MgCl2 and 1 mg / ml X-gal. The number of SA-β-Gal positive cells was determined in five randomly selected fields of view and calculated as a ratio to the control without added maturation stimulants.

[0148] [RNA Extraction and RT-PCR-based Quantification]

[0149] For endometrial cell populations (pESC, Ishikawa cells, EM1 cells) and cells obtained from removed uteruses, genes were extracted from cultured cells using Isogen II (manufactured by Nippon Gene Co., Ltd.). mRNA was reverse transcribed using a ReverTra Ace qPCR RT Kit (manufactured by Toyobo Co., Ltd.), and the resulting cDNA was amplified by qPCR using PowerUP SYBR Green Master Mix (manufactured by Thermo Fisher Scientific Co., Ltd.). The Ct value of each gene was used as the expression level, and the expression ratio was calculated using the expression level without added PQQ as a control.

[0150] [Evaluation of gene expression in mouse endometrial epithelium]

[0151] Starting one week before the experiment, 10-week-old female C57BL / 6 mice were given PQQ (2 mg / day) via drinking water. In these mice, day 0 was designated as the day of estrus, and pseudopregnancy was performed (adult female mice were mated with male mice that had undergone vasectomy at a ratio of 3:1, and vaginal plugs were confirmed the next morning; mice with vaginal plugs were used). The uterus was removed 4 and 5 days later as specimens. For these specimens, uterine tissue was homogenized and lysed with lysis buffer to evaluate gene expression (Example 4-1). In addition, paraffin sections were prepared directly from the uterus, and FOXO1-positive cells were immunostained (Example 4-2).

[0152] [Evaluation of implantation in mouse uterus]

[0153] Starting one week before the experiment, 10-week-old female C57 / BL6 mice were given PQQ (2 mg / day) via drinking water. In this mouse, day 0 was designated as the day of estrus, and a pseudopregnancy treatment was performed (adult female mice were mated with male mice that had undergone vasectomy at a ratio of 3:1, and the vaginal plug was confirmed the next morning; mice with vaginal plugs were used). The next day, two-cell stage embryos were transferred into the fallopian tubes, and the uterus was retrieved 10 days later to count the number of implanted embryos. Mice that did not drink PQQ were used as controls, and the implantation rate was calculated.

[0154] [Experiment 1. Cytotoxicity test]

[0155] 1 pM to 1 mM (common ratio 10) of PQQ was added to pESC. The number of viable cells was evaluated using NADH as an indicator, and the number of dead cells was evaluated using LDH as an indicator. The results are shown in Figure 1.

[0156] As a result, no significant increase in dead cells or decrease in viable cells was observed in any of the evaluated concentration regions. That is, it was determined that the effects of reduced expression of various factors caused by cell death in the following experimental examples were negligible.

[0157] [Experimental Example 2-1. Decidualization Promotion Assay of pESC]

[0158] PQQ was added to pESC at concentrations of 1 nM to 100,000 nM (common ratio 10) simultaneously with the maturation stimulating component (MPA+Db). The group without the maturation stimulating component was designated as the control (CM), and the group with only the maturation stimulating component added was designated as (MPA+Db). The group with PQQ added simultaneously with the maturation stimulating component (page 10 / 13, CN 122295102 A) was designated as "PQQ+DM".

[0159] RNA extraction and RT-PCR quantification were performed in each group to evaluate the expression of genes related to decidualization. The results are shown in Figure 2.

[0160] The results showed that at any concentration of PQQ, the representative genes of decidualization, RPL, IGFBP1, and FOXO1, were all significantly upregulated.

[0161] In summary, PQQ significantly promoted decidualization of human primary endometrial stromal cells even at the lowest concentration of 1 nM.

[0162] [Experimental Example 2-2. Epithelial Differentiation Promotion Assay of Ishikawa and EM1 Cells]

[0163] PQQ was added to Ishikawa and EM1 cells at concentrations of 1 nM to 100,000 nM (common ratio 10) simultaneously with the maturation stimulating component (Db). The group without the maturation stimulating component was designated as the control (CM). The group with only the maturation stimulating component was designated as (Db), and the group with PQQ added simultaneously with the maturation stimulating component was designated as "PQQ+D".

[0164] RNA extraction and RT-PCR quantification were performed in each group to evaluate the expression of genes related to epithelial differentiation. The results are shown in Figure 3.

[0165] The results showed that in Ishikawa and EM1 cells, at any concentration of PQQ, the representative genes for epithelial differentiation, LIF and COX2, were significantly upregulated.

[0166] In summary, PQQ can significantly promote epithelial differentiation of human endometrial epithelial cell lines and human endometrial glandular epithelial stromal cell lines even at the lowest concentration of 1 nM.

[0167] As shown in Experiments 2-1 and 2-2, PQQ significantly promotes decidualization in primary endometrial stromal cells and promotes differentiation in endometrial epithelial cell lines and endometrial glandular epithelial cell lines. Therefore, it is determined that PQQ has the function of promoting endometrial tissue maturation and assisting implantation.

[0168] [Experiment 3-1. Inhibition test of senescence-related β-galactosidase accompanying pESC decidualization]

[0169] 1 nM to 100 nM (common ratio 10) of PQQ was added to pESC simultaneously with the maturation stimulating component (MPA+Db). Using no maturation stimulating component as the control (Ctrl), the group with only the maturation stimulating component added was denoted as (DM), the group containing only PQQ was denoted as (PQQ), and the group with PQQ added simultaneously with the maturation stimulating component was denoted as "DM+PQQ".

[0170] In each group, senescence-related β-galactosidase was stained, and the stained cell rate (number of stained cells / total number of cells) was calculated. The results are shown in Figure 4.

[0171] The results showed that maturation stimulation led to a significant increase in senescence-related β-galactosidase, but at any concentration of PQQ, senescence-related β-galactosidase was significantly inhibited.

[0172] [Experiment 3-2. Inhibition of senescence gene p53 and promotion of anti-aging gene accompanied by pESC decidualization]

[0173] 1 nM to 1000 nM (common ratio 10) of PQQ was added to pESC simultaneously with the maturation stimulating component (MPA+Db). Using the group without the maturation stimulant as the control (CM), the group with only the maturation stimulant added was designated as (DM), and the group with PQQ added in conjunction with the maturation stimulant was designated as "DM+PQQ".

[0174] The expression levels of the aging gene p53 and the anti-aging gene SIRT1 in each group were measured, and the relative expression ratios relative to the control were calculated. The results are shown in Figure 5.

[0175] As a result, it was found that maturation stimulation led to a significant increase in the expression of the aging gene p53, but at any concentration of PQQ, the expression of p53 was significantly inhibited to the same level as the control. In addition, it was found that maturation stimulation led to the inhibition of the expression of the anti-aging gene SIRT1, but at any concentration of PQQ, the expression of SIRT1 was significantly enhanced to the same level as the control.

[0176] As shown in the results of Experiments 3-1 and 3-2, in primary endometrial stromal cells, PQQ inhibited the aging factors (aging-related β-galactosidase, aging gene p53) caused by decidualization stimulation and enhanced the anti-aging factor (anti-aging gene SIRT1) on pages 11 / 13 of the anti-aging manual. Therefore, it was determined that PQQ has the function of inhibiting the aging of endometrial tissue during the maturation process.

[0177] In summary, this indicates that PQQ can promote the maturation of endometrial tissues (stromal, epithelium, glandular epithelium) in vitro while inhibiting cell senescence. Therefore, it is judged that PQQ promotes and assists normal endometrial maturation, which helps to promote and assist implantation.

[0178] [Experimental Example 4-1. Gene expression of endometrial epithelium in mice that drank PQQ (RT-PCR) test]

[0179] The expression of genes related to decidualization and epithelial differentiation in the entire uterus of mice that drank PQQ was evaluated. The results showed that the expression of genes related to decidualization and epithelial differentiation, LIF1, ITGB3, FOXO1, BMP2, and COX2, was significantly enhanced, as in the in vitro test results (Figure 6).

[0180] [Experiment 4-2. Gene expression and immunostaining of endometrial epithelium in mice that drank PQQ]

[0181] The expression of FOXO1, a representative gene related to epithelial differentiation, in the uterus of mice that drank PQQ was evaluated. The results showed a significant increase in FOXO1-positive cells, indicating that epithelial differentiation occurred even at the cellular level in the uterus (Figure 7).

[0182] As shown in the results of Experiments 4-1 and 4-2, the characteristic gene for epithelial differentiation was highly expressed and the number of epithelial differentiated cells increased in the uterus of mice that drank PQQ. Therefore, it was determined that PQQ can promote epithelial differentiation of the endometrium not only in vitro but also in animal models, and assist and promote the implantation of the endometrium.

[0183] [Experiment 5. Implantation rate evaluation experiment in mice that drank PQQ]

[0184] The implantation rate of transplanted embryos in the uterus of mice that drank PQQ was evaluated. The results showed that the implantation rate was significantly increased compared with mice that did not drink PQQ (Figure 8).

[0185] According to the results of Experiments 1 to 5, PQQ has the effect of maturing endometrial stromal cells, epithelial cells and glandular epithelial cells that play an important role in implantation. At the same time, it also has the effect of inhibiting cell senescence caused by overactivation, which is ideal for promoting and assisting implantation. As a result of this effect, the embryo implantation rate is increased.

[0186] [Experimental Example 6. Gene Expression Assay for Cell Fusion]

[0187] For placental trophoblast cells, with and without the addition of foscoline (0.5 μM, manufactured by Cayman Chemical Co., Ltd., hereinafter referred to as "FSK") as a fusion stimulus, 10 nM of PQQ was simultaneously added to the culture medium [with the addition of 4% knockout serum substitute (Thermo Fisher Scientific), 0.1 mM 2-mercaptoethanol (manufactured by Fujifilm and Kojun Pharmaceutical Co., Ltd.), 0.3% bovine serum albumin (manufactured by Fujifilm and Kojun Pharmaceutical Co., Ltd.), 1% ITS-X supplement (manufactured by Fujifilm and Kojun Pharmaceutical Co., Ltd.), 2.5 μM Y27632 (manufactured by Fujifilm and Kojun Pharmaceutical Co., Ltd.), 50 ng / mL The cells were cultured in Dulbecco modified Eagle medium / F12 (DMEM / F12) (1:1) medium containing epithelial growth factor (manufactured by Fujifilm and Kodenki Chemical Co., Ltd.), antibiotics, and antifungal drugs. The fusion behavior was compared between the PQQ-free and PQQ-added states.

[0188] RNA extraction and RT-PCR quantification were performed under each condition to evaluate the gene expression of Syncytin 2 and PAPPA as fusion markers. The results are shown in Figure 9.

[0189] Figure 9 shows the state without fusion stimulation (Ctrl), PQQ-only (PQQ), fusion-induced trophoblast cells (FSK), and the state with PQQ added to the fusion stimulation (FSK+PQQ).

[0190] The results indicate that the gene expression of Syncytin 2 and PAPPA as fusion markers significantly increased with the addition of PQQ.

[0191] In summary, PQQ significantly promotes cell fusion.

[0192] [Experimental Example 7. Cell Fusion Promotion Assay]

[0193] Subsequently, human trophoblast stem cells treated with fusion-stimulated FSK were fixed with methanol and incubated with anti-E-cadherin antibody (1:200, #3195, CST) and AlexaFluor 488-labeled goat anti-mouse antibody (Thermo Fisher manual, pages 12 / 13, CN 122295102 A Scientific) to recognize the cell surface. The staining photographs at this time are shown in Figure 10.(Experimental Example 7-1)

[0194] Cell nuclei were counterstained with 4',6-diamino-2-phenylindole dihydrochloride (DAPI). The number of nuclei and the total number of nuclei in syncytiotrophoblast cells (fused cells) were counted in five randomly selected microscopic fields for each sample and calculated as a fusion index [(number of nuclei in fused cells / total number of nuclei) × 100] in three independent experiments. The resulting increase in the number of fused cells is shown in Figure 11. The use of PQQ was confirmed to increase the number of fused cells. The invention provides a preparation capable of promoting normal decidualization of the endometrium. A promoter or improver for decidualization of the endometrium, containing an effective amount of pyrroloquinoline quinone or a salt thereof, is prepared. Furthermore, a promoter or improver for cell fusion, containing an effective amount of pyrroloquinoline quinone or a salt thereof, is prepared. Abstract.

Claims

1. An agent that promotes or improves endometrial decidualization, comprising an effective amount of pyrroloquinoline quinone or its salt.

2. An anti-aging agent for the endometrium, comprising an effective amount of pyrroloquinoline quinone or its salt.

3. The medicament according to claim 2, wherein, The anti-aging effect of the endometrium is to inhibit the transformation of endometrial cells into senescent cells.

4. The pharmaceutical preparation according to claim 2, wherein, The anti-aging process of the endometrium involves removing senescent cells from the endometrium.

5. A placental trophoblast cell fusion promoter or improver, comprising an effective amount of pyrroloquinoline quinone or its salt.

6. A fertility enhancer containing an effective amount of pyrroloquinoline quinone or its salt, which enhances fertility by promoting or improving endometrial decidualization and / or promoting or improving placental trophoblast cell fusion.

7. The pharmaceutical agent according to claim 6, wherein, The improvement in fertility refers to improved implantation or continued implantation.

8. The medicine according to any one of claims 1 to 7, wherein it is ingested for at least 7 days to 21 days after the first day of menstruation.

9. The medicine according to any one of claims 1 to 7, wherein it is ingested for at least 14 days to 35 days after the first day of menstruation.

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