Application of ergothioneine in preparation of preparation for improving ovarian reserve function

Abstract

The invention relates to application of ergothioneine in preparation of a preparation for improving an ovarian reserve function. The invention creatively discovers that the ergothioneine has the function of improving the ovarian reserve function, and the obvious clinical value of the ergothioneine for people with the serum AMH level of 1.0-3.0 ng / mL in the key'window phase 'is defined for the first time. The ergothioneine not only can improve the serum AMH level and the sinus follicle count (AFC), but also can significantly reduce the abnormally increased FSH and LH levels. Meanwhile, aiming at bone loss caused by ovarian reserve function decline, the ergothioneine shows an excellent bone quality protection effect, the bone density can be improved, and the thickness and number of bone trabecula can be increased. In addition, the ergothioneine can improve climacteric symptoms, effectively maintain the nutritional state of target organs such as uterus, delay atrophy and regulate menstrual cycle disorder.

Description

[0001] This application claims priority to patent application number 202610157730.5 (the earlier application was filed on February 4, 2026, and was entitled "Use of Ergothionein in the Preparation of a Preparation for Improving Ovarian Reserve Function"). Technical Field

[0002] This invention relates to the field of pharmaceutical technology, and in particular to the application of ergothioneine in the preparation of agents that improve ovarian reserve function. Background Technology

[0003] Ovarian reserve refers to the quantity and quality of primordial follicles in the ovarian cortex, reflecting a woman's reproductive potential. With increasing social and environmental pressures, a faster pace of life, and delayed age of first childbearing, the incidence of diminished ovarian reserve (DOR) and premature ovarian insufficiency (POI) is rising annually and showing a trend towards affecting younger women. Anti-Müllerian hormone (AMH) is the "gold standard" for assessing ovarian reserve, and its level is positively correlated with antral follicle count (AFC). Clinically, when serum AMH levels are in the range of 1.0-3.0 ng / mL, it often indicates that a woman's ovarian function is entering a critical "window period" of decline. Women in this stage often experience menstrual irregularities (shortened or prolonged cycles, abnormal menstrual flow), endocrine disorders (abnormally elevated FSH and LH, fluctuating and declining E2 levels), and if not addressed promptly, will accelerate the transition to menopause.

[0004] The decline in ovarian function is not merely a localized problem within the reproductive system, but can trigger a systemic health crisis. Among these crises, the decrease in estrogen levels leads to dysfunction of the hypothalamic-pituitary-ovarian axis (HPO axis), thereby affecting overall metabolism. One of the most significant complications is postmenopausal osteoporosis (PMOP). Estrogen has a protective effect on bones, inhibiting osteoclast activity. When ovarian function declines and estrogen withdrawal occurs, bone resorption exceeds bone formation, leading to rapid bone loss and damage to bone microstructure (such as a decrease in the number and thickness of trabeculae, and increased separation), thus greatly increasing the risk of fragility fractures. Current medical research urgently needs to find a comprehensive intervention that can both improve ovarian function and effectively prevent accompanying bone loss.

[0005] Currently, treatments for decreased ovarian reserve and perimenopausal syndrome mainly include: Hormone replacement therapy (HRT): Although it can effectively relieve symptoms and prevent bone loss, long-term use carries the potential risk of increasing breast cancer, endometrial cancer and thromboembolic diseases, and has many contraindications and poor adherence.

[0006] Adjunctive drugs, such as dehydroepiandrosterone (DHEA) and coenzyme Q10, have been used to improve ovarian function to some extent, but they are not very targeted and lack sufficient evidence-based medical evidence for their precise improvement effect on specific AMH value groups (such as 1.0-3.0 ng / mL).

[0007] Osteoporosis treatment drugs, such as bisphosphonates (e.g., alendronate sodium), although they have a protective effect on bones, cannot improve ovarian endocrine function, and long-term use may lead to side effects such as osteonecrosis of the mandible.

[0008] Ergothioneine (EGT) is a natural histidine betaine derivative known for its excellent antioxidant, anti-inflammatory, and cell-protective properties. Unlike traditional antioxidants (such as coenzyme Q10 and vitamin E) that passively diffuse, ergothioneine possesses a unique "privileged channel" in vivo—the specific transport protein OCTN1 (SLC22A4). Studies have shown that OCTN1 receptors are highly expressed in ovarian tissue and oocytes, allowing ergothioneine to penetrate the cell membrane via this transport protein and further target and accumulate in the cell's "energy factories"—mitochondria. Given that the core mechanism of oocyte aging lies in mitochondrial dysfunction and oxidative damage, ergothioneine's ability to precisely target mitochondria, scavenge free radicals, and maintain energy metabolism homeostasis gives it an irreplaceable and unique advantage in improving ovarian reserve function. Current research on ergothioneine focuses primarily on its skin anti-aging, neuroprotective, and cardiovascular protective effects. However, there are no reports on the application of ergothionein in improving ovarian reserve function in people with AMH 1.0-3.0 ng / mL, and there is also a lack of systematic research on its ability to simultaneously improve bone microstructure damage (increase bone density and increase the number of trabeculae) caused by ovarian function decline and regulate HPO axis hormone levels. Summary of the Invention

[0009] To address the aforementioned technical problems, this invention provides the application of ergothionein in the preparation of agents that improve ovarian reserve function.

[0010] To achieve this objective, the present invention adopts the following technical solution: In a first aspect, the present invention provides the application of ergothionein in the preparation of preparations for improving ovarian reserve function.

[0011] In this invention, the efficacy indicator of ovarian reserve function is the change in serum anti-Müllerian hormone (AMH) level; the auxiliary observation indicators are sex hormones (follicle-stimulating hormone FSH, luteinizing hormone LH, estrogen E2, and testosterone T).

[0012] The test results provided by this invention demonstrate that ergothioneine can regulate the hypothalamus-pituitary-ovarian (HPO) axis, reduce cortisol pressure, and improve sleep to increase the pulsatile secretion of endogenous gonadotropins, thereby increasing serum anti-Müllerian hormone levels and antral follicle count.

[0013] Preferably, the formulation is suitable for individuals with serum anti-Müllerian hormone levels of 1.0-3.0 ng / mL (e.g., 1.0 ng / mL, 1.5 ng / mL, 2 ng / mL, 2.5 ng / mL, 3 ng / mL, etc.).

[0014] Secondly, this invention provides the application of ergothionein in the preparation of a formulation to improve bone loss caused by declining ovarian reserve function.

[0015] In this invention, ergothioneine exhibits excellent bone quality protection effects, increasing bone density and the thickness and number of trabeculae.

[0016] Thirdly, this invention provides the application of ergothionein in the preparation of formulations that improve menopausal symptoms caused by declining ovarian reserve.

[0017] In this invention, ergothioneine can improve menopausal symptoms, effectively maintain the nutritional status of target organs such as the uterus, delay atrophy, reduce FSH / LH, and maintain E2 / P balance.

[0018] Fourthly, the present invention provides the use of ergothionein in the preparation of preparations for regulating sex hormone levels, wherein the sex hormone is selected from any one or a combination of at least two of estrogen, follicle-stimulating hormone, luteinizing hormone or testosterone.

[0019] Fifthly, this invention provides the application of ergothioneine in the preparation of preparations for regulating menstrual cycle disorders.

[0020] Preferably, the formulation further contains pharmaceutically acceptable excipients.

[0021] Preferably, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

[0022] Preferably, the dosage form of the preparation includes tablets, granules, capsules, powders, pills, solutions, syrups, emulsions, suspensions, ointments, or microspheres.

[0023] Compared with the prior art, the present invention has at least the following beneficial effects: 1. Precisely defines and significantly improves ovarian reserve function in women within the "AMH 1.0-3.0 ng / mL" window. This invention overcomes the limitations of traditional ovarian maintenance agents with their vague target population, creatively discovering that ergothioneine has a significant "therapeutic window" effect on women with serum AMH levels in the specific range of 1.0-3.0 ng / mL. Clinical trial data show that after taking the formulation of this invention, serum AMH levels in this population significantly rebounded, and antral follicle count (AFC) increased significantly. This indicates that this invention can effectively activate ovarian function on the verge of decline, delay the ovarian aging process, and provide a precise intervention plan for women at this stage.

[0024] 2. It has a dual protective effect on both the ovaries and bones, which is superior to single-target drugs. This invention provides a systematic solution to the bone loss problem that often accompanies ovarian dysfunction.

[0025] 3. Repairing the HPO axis feedback mechanism and restoring endocrine homeostasis. This invention confirms that ergothioneine has excellent neuroendocrine regulatory effects. It can repair the imbalanced hypothalamic-pituitary-ovarian axis (HPO axis) and fundamentally alleviate perimenopausal and premature ovarian failure-related symptoms such as hot flashes, sweating, and mood swings by improving the negative feedback regulatory mechanism.

[0026] 4. Significant target organ protection and improved quality of life. This invention demonstrates a direct protective effect on estrogen target organs, such as the uterus.

[0027] 5. High safety profile, suitable for long-term use. Compared with traditional hormone replacement therapy (HRT), the ergot thiosulfate used in this invention, being a naturally derived active ingredient, does not directly supplement exogenous hormones and can be used for daily intervention in women with diminished ovarian reserve and perimenopause. Detailed Implementation

[0028] To facilitate understanding of the present invention, the following embodiments are provided. Those skilled in the art should understand that these embodiments are merely illustrative and should not be construed as limiting the scope of the invention.

[0029] Example 1 Effects of ergothionein on bone metabolism and endocrine function in ovariectomized (OVX) mice Test method: 1. Laboratory animals Animal strain: C57BL / 6 mice, age / weight: 8 weeks, 18-22g, female; Environmental system: SPF-grade animal housing, controlled temperature: 20-26℃ (daily temperature difference ≤4℃), controlled humidity: 40%-70%, air exchange rate: ≥15 times / h, 100% fresh air, working illuminance: ≥200 Lx, animal illuminance: 15-20 Lx, lighting time: 12 h / 12 ​​h (07:00~19:00) with alternating light and dark; Feed Name: SPF Rat and Mouse Maintenance Compound Feed (Maintenance), Manufacturer: Keao Xieli (Tianjin) Feed Co., Ltd., Production License Number: SCXK (Jin) 2020-0004.

[0030] 2. Experimental Procedure 2.1 Modeling Stage Adaptation: All mice were acclimatized for 5 days, then weighed and randomly grouped. Surgery: After acclimatization for at least 5 days, the target animals were surgically modeled once their weight was within the range. Except for the blank control group, all mice underwent respiratory anesthesia. The main surgical steps were as follows: After fixing the mice, removing hair, preparing the skin, and disinfecting the target area, the skin, fascia, and muscle layers were cut open with tissue scissors. Fat was separated to expose granular ovarian tissue. The fallopian tubes were ligated with sutures, and both ovaries were completely removed with tissue scissors to simulate the physiological state after ovarian failure (i.e., perimenopause / postmenopause). After the surgery, the wound was sutured layer by layer and disinfected again with povidone-iodine. The mice were then returned to their cages and allowed free access to food. Low-dose penicillin was administered intramuscularly for three consecutive days to prevent infection. The blank control group received no intervention.

[0031] One week after surgery, except for the sham control group, the other mice were fed a low-calcium, low-protein diet for one week, and gavage intervention began one week later.

[0032] 2.2 Treatment Phase The modeled mice were randomly divided into groups of 8. (1) Blank control group (equal volume of water administered by gavage) (2) Model group (equal volume of water administered by gavage) (3) Positive group (alendarum sodium 1.53 mg / kg) (4) Low-dose ergothioneine group (10 mg / kg) (5) High-dose ergothioneine group (30 mg / kg) Animals in each group were given the drug and dosage according to their respective drugs. Except for the positive group, which was given once a week, the other groups were given the drug once a day, five times a week, for a total of 4 weeks. Two mice were taken for process tracking, and the remaining 6 mice were given the drug for another 8 weeks before being tested. The final statistics were made after the model stabilized.

[0033] 3. Indicator Testing 3.1 Bone quality indicators: Micro-CT examination of bone tissue morphology After the last gavage, the mice were fasted for 12 hours, weighed, anesthetized, and blood was collected. The mice were then euthanized, and the serum was separated and frozen at -20°C for later use. After thoroughly dissecting the muscle tissue around the tibia, one femur from each side of the mice was fixed in 40 g / L paraformaldehyde, and the other was frozen at -80°C for later use. The femur specimens were scanned using a small animal Micro CT imaging system to obtain raw data projection images. Bone density, trabecular thickness, and trabecular number were obtained through data analysis, and 3D visualization of the trabecular bone was performed.

[0034] 3.2 Bone metabolism indicators: Type I procollagen aminoterminal peptide (P1NP) Type I collagen cross-linked C-terminal peptide (CTX) Osteocalcin (OC / BGP) 3.3 Menopausal indicators: Uteruses were collected from each group of animals and weighed (uterine weight: atrophic changes, directly related to estrogen deficiency), and the levels of estrogen (E2), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were tested.

[0035] 3.4 Oxidative stress indicators: IL-6, TNF-α, SOD activity, MDA.

[0036] 4. Experimental equipment 5. Reagent kit 6. Sample Collection and Preprocessing Tissue sample handling: (1) Weigh the animal tissue and add 9 times the volume of homogenizing medium (0.85% physiological saline) at a ratio of weight (mg): volume (μL) = 1:9. (2) Mechanical homogenization was performed using a high-speed tissue homogenizer; (3) Place the homogenate in a benchtop high-speed refrigerated centrifuge and centrifuge at 2500 rpm for 10 minutes. Collect the supernatant for later use.

[0037] 7. Experimental Procedure (ELISA Double Antibody Sandwich Method) Sample addition and incubation: Set up standard wells and test sample wells in the microplate. Add 50 μL of different concentrations of standard to the standard wells; add 40 μL of sample diluent to the sample wells, followed by 10 μL of the test sample (final dilution is 5-fold). Then add 100 μL of HRP-labeled detection antibody (i.e., enzyme-labeled reagent) to each well, seal the plate with sealing film, and incubate at 37°C for 60 minutes.

[0038] Washing: Discard the liquid inside the well, pat dry on absorbent paper, add washing solution using a fully automatic plate washer, let stand for 30 seconds, then drain, repeat washing 5 times to remove unbound components.

[0039] Color development: Add 50 μL of color developer A and 50 μL of color developer B to each well in sequence, gently shake to mix, and develop at 37°C in the dark for 15 minutes.

[0040] Termination and measurement: Add 50 μL of stop solution to each well (the color changes from blue to yellow), and immediately measure the absorbance (OD value) of each well at a wavelength of 450 nm using a microplate reader.

[0041] 8. Data Processing and Result Determination Curve fitting: Plot a standard curve with the concentration of the standard as the x-axis and the OD value as the y-axis, and obtain the linear regression equation.

[0042] Concentration calculation: The corresponding concentration is calculated in the regression equation based on the OD value of the sample, and then multiplied by the dilution factor of the sample to obtain the actual concentration of the target protein or hormone in the sample.

[0043] The results of the 8-week tests on bone quality, bone metabolism, menopause, and oxidative stress indicators are shown in Tables 1, 2, 3, and 4, respectively.

[0044] Table 1 Bone quality indicators Table 2 Bone Metabolism Indicators Table 3 Menopausal Indicators Table 4 Oxidative Inflammation Indicators The results show: (1) Bone quality and bone metabolism indicators showed that ergothioneine has excellent bone quality protection effect, can increase bone density, increase the thickness and number of trabeculae, and increase type I procollagen amino-terminal peptide, decrease type I cross-linked C-terminal peptide of collagen, and decrease osteocalcin in bone metabolism indicators, indicating that ergothioneine can improve bone loss caused by ovarian dysfunction.

[0045] (2) Menopausal indicators showed that the uterine atrophy was reduced and the uterine weight was higher in the ergothioneine group than in the OVX model group, indicating that ergothioneine can effectively maintain the nutritional status of target organs and delay the atrophy of reproductive organs. Serum E2 levels in the ergothioneine group increased, indicating that ergothioneine can regulate endogenous estrogen levels and maintain endocrine homeostasis under impaired ovarian function. Serum FSH levels in the ergothioneine group decreased significantly (which spikes after OVX), indicating that ergothioneine significantly reduces abnormally elevated gonadotropins and repairs the imbalance of the hypothalamus-pituitary-ovarian axis (HPO axis) through negative feedback regulation. Serum LH levels in the ergothioneine group decreased significantly, indicating that ergothioneine can synergistically regulate pituitary secretion and improve neuroendocrine disorders caused by hormonal fluctuations. These results indicate that ergothioneine can improve menopausal symptoms.

[0046] (3) Oxidative stress test results showed that ergothionein blocked the MDA-mediated cell membrane lipid peroxidation chain reaction by scavenging reactive oxygen species (ROS) and inducing the recovery of SOD activity, thus protecting the structural integrity of ovarian granulosa cells at the molecular level.

[0047] Example 2 Human clinical trials Test method: 1. Experimental materials and subject screening Experimental product and dosage: Ergothioneine capsules (the main ingredient is ergothioneine), daily supplementation of 120mg of ergothioneine, batch number: P776678, provided by Jiangsu Jinsan Biotechnology Co., Ltd. All products used in this experiment came from the same batch number.

[0048] Subject inclusion criteria: Women aged 35-45 (inclusive); It manifests as menstrual disorders, including infrequent or frequent menstruation, abnormal cycles, amenorrhea, abnormal menstrual flow, or dysmenorrhea; Biochemical analysis confirmed that the level of anti-Müllerian hormone (AMH) was between 1.0 ≤ AMH < 3 ng / mL; Exclusion criteria: Excluded patients with serious primary diseases such as cardiovascular, liver and kidney diseases, those who have previously undergone oophorectomy, those with polycystic ovary syndrome or other endocrine disorders affecting ovulation, as well as pregnant and lactating women.

[0049] 2. Experimental Design and Dosing Protocol Research model: A single-center, self-controlled pre- and post-treatment human trial design was adopted; Sample size: A total of 40 eligible subjects were included; Experimental period: Three consecutive menstrual cycles of use.

[0050] 3. Experimental Procedure and Evaluation Indicators Intervention process: Screening and baseline period: Subjects are screened and qualified subjects are recorded for baseline data, including AMH and sex hormones; Implementation period: Subjects took the product continuously at the prescribed dosage, and follow-up tests were conducted on days 2-4 of the first, second, and third menstrual cycles.

[0051] Evaluation period: After the third menstrual cycle ends (post-group visit), the final effectiveness evaluation and safety check will be conducted.

[0052] Core evaluation indicators: Efficacy indicator: Changes in serum anti-Müllerian hormone (AMH) levels; Auxiliary observation indicators: sex hormones (follicle-stimulating hormone FSH, estrogen E2); Subjective symptoms: Menstrual cycle information was recorded and assessed using the modified Kupperman score and the Pittsburgh Sleep Quality Index (PSQI).

[0053] The menstrual cycle scoring table is shown in Figure 5. The scoring method is to add up the scores of each indicator.

[0054] Table 5 Menstrual Status Record Scoring Sheet The Modified Kupperman Scale is a menopausal / perimenopause symptom assessment scale (see Table 6) that objectively reflects the severity of menopausal symptoms in women. Scoring method: The score for each symptom is the base score multiplied by the severity score. The scores for all symptoms are added together, with a total score ranging from 0 to 63.

[0055] Table 6 Modified Kupperman Rating Scale The Pittsburgh Sleep Quality Index (PSQI) has a total score of 0-21 and is used to quantitatively assess sleep quality over the past month. The scale has 24 items, of which 18 are self-report items that are scored. These 18 items make up 7 dimensions, with each dimension scored from 0 to 3 points, for a total score of 0-21 points. The lower the score, the better the sleep quality.

[0056] Table 7. Pittsburgh Sleep Quality Index (PSQI) Table 8. Meaning and Scoring Methods of Each Dimension of PSQI 4. Safety Evaluation Monitoring indicators: The vital signs, physical examination results and laboratory indicators (complete blood count and blood biochemistry) of the subjects were recorded throughout the trial.

[0057] 5. Efficacy evaluation results 5.1 Analysis of Anti-Müllerian Hormone (AMH) Levels Anti-Müllerian hormone (AMH) is a glycoprotein hormone secreted by small follicles in the ovary, and its core function reflects ovarian reserve. Table 9 shows that the average baseline AMH level was 1.79 ng / mL. By the third menstrual cycle after taking the medication, the AMH level was 2.5 ng / mL, a significant increase of 36.26% compared to baseline. p =0.0292), indicating that ovarian reserve function has been effectively improved.

[0058] Table 9. Anti-Müllerian Hormone (AMH) Levels After 3 Menstrual Cycles of Ergothioneine Capsule Intake 5.2 Analysis of Follicle-Stimulating Hormone (FSH) Levels Follicle-stimulating hormone (FSH) is secreted by the pituitary gland and directly regulates the growth and development of ovarian follicles, making it a core indicator for assessing ovarian function. Table 10 shows that the baseline average FSH level was 8.22 mIU / mL. By the third menstrual cycle after taking the medication, the FSH level was 7.05 mIU / mL, a significant decrease of 16.21% compared to the baseline level. p =0.0317), the regulatory function of follicle growth and development was optimized.

[0059] Table 10. Follicle-stimulating hormone (FSH) levels after 3 menstrual cycles of taking ergothioneine capsules 5.3 Analysis of estradiol (E2) levels Estradiol (E2) is the main estrogen secreted by ovarian granulosa cells, reflecting follicular activity and endometrial development. Table 11 shows that the baseline average estradiol level was 46.00 ng / mL. By the third menstrual cycle, the E2 level was 63.46 ng / mL, a significant increase of 42.79% compared to baseline. p =0.0305), indicating that follicular activity and endometrial development were significantly improved.

[0060] Table 11 Estradiol (E2) levels after 3 menstrual cycles of taking ergothioneine capsules 5.4 Improved Kupperman Rating Analysis The Modified Kupperman Score is a menopausal / perimenopause symptom assessment scale that objectively reflects the severity of menopausal symptoms in women. Table 12 shows that the baseline average Modified Kupperman Score was 5.4. After 3 menstrual cycles of use, the score decreased to 1.9, a significant reduction of 64.00% compared to baseline. p <0.0001). The test results show that ergothioneine capsules can effectively relieve various menopausal discomfort symptoms, and the improvement effect becomes more and more significant with the extension of the treatment period.

[0061] Table 12 Improved Kupperman Score after 3 Menstrual Cycles of Ergothioneine Capsule Intake 5.5 Analysis of Pittsburgh Sleep Quality Index (PSQI) Scale Scores The Pittsburgh Sleep Quality Index (PSQI) has a total score of 0-21 and is used to quantitatively assess sleep quality over the past month. Table 13 shows that the baseline average PSQI level was 6.9. After 3 menstrual cycles of use, the PSQI level was 5.5, a decrease of 22.89% compared to baseline. p <0.0001).

[0062] Table 14 shows that after 3 menstrual cycles of taking ergothioneine capsules, the sleep disorder score of the participants was significantly lower than before the trial. The baseline average score for the sleep disorder dimension was 2.9 points, and after 3 menstrual cycles, the score was 1.5 points, a decrease of 46.83% compared to baseline. p <0.0001).

[0063] Based on the analysis of the Pittsburgh Sleep Quality Index (PSQI) scores, taking ergothioneine capsules for 3 menstrual cycles can effectively improve sleep quality and alleviate sleep disorders.

[0064] Table 13 Pittsburgh Sleep Quality Index (PSQI) scores after 3 menstrual cycles of taking ergothioneine capsules. Table 14. Pittsburgh Sleep Quality Index (PSQI) - Sleep Disorders Dimension Scores after 3 Menstrual Cycles of Ergothioneine Capsule Use 5.6 Menstrual Record Scoring Analysis The menstrual cycle record score is a comprehensive quantitative score based on dimensions such as cycle regularity, menstrual period length, menstrual flow, cycle pain, and cycle predictability. Table 15 shows that the baseline average score for the menstrual cycle record score was 18.9 points. After three menstrual cycles of use, the score was 21.0 points, a significant increase of 12.20% compared to baseline. p =0.0006). In summary, the analysis results indicate that taking ergothioneine capsules for three menstrual cycles can effectively improve and optimize overall menstrual health.

[0065] Table 15. Menstrual cycle records and scores after taking ergothioneine capsules for 3 menstrual cycles. The results showed that after 3 menstrual cycles of taking ergothioneine capsules, the core indicators of ovarian function in the participants were significantly improved. Specifically, anti-Müllerian hormone (AMH) levels significantly increased by 36.26% (p=0.0292), indicating an effective improvement in ovarian reserve; follicle-stimulating hormone (FSH) levels significantly decreased by 16.21% (p=0.0317), optimizing the regulation of follicle growth and development; and estradiol (E2) levels significantly increased by 42.79% (p=0.0305), significantly improving follicular activity and endometrial development. The synergistic effect of FSH and E2 is a relatively ideal response pattern in the improvement of ovarian function, suggesting that ergothioneine may play a beneficial regulatory role in basal ovarian function in individuals with low AMH levels through endocrine regulation, improvement of the ovarian microenvironment, and reduction of oxidative stress.

[0066] Modified Kupperman score results showed that ergothioneine capsules sustained and significantly reduced the severity of menopausal symptoms in trial participants. After 3 menstrual cycles of use, the score decreased significantly by 64.00% (p<0.0001), indicating that ergothioneine capsules can effectively relieve various menopausal discomfort symptoms (hot flashes, night sweats, mood swings), and the improvement effect becomes more significant with the extension of the use period.

[0067] The Pittsburgh Sleep Quality Index (PSQI) results showed that after 3 menstrual cycles of use, the total score decreased significantly by 22.89% (p<0.0001). Among them, the improvement in the sleep disorder dimension score was more significant, decreasing significantly by 46.83% after 3 menstrual cycles (p<0.0001), indicating that ergothioneine capsules can effectively improve sleep quality and alleviate sleep disorders.

[0068] The menstrual status record score results showed that after taking ergothioneine capsules for 3 menstrual cycles, the menstrual status of the participants was significantly improved, with a significant increase in score of 12.20% (p=0.0006), indicating that ergothioneine capsules can effectively improve menstrual status (including cycle regularity, menstrual flow, dysmenorrhea, etc.).

[0069] 6. Safety Performance A total of 40 participants were enrolled in this trial. Throughout the study, no abnormalities were observed in the participants' safety indicators, and no adverse reactions related to the test product were observed. This fully demonstrates the excellent safety profile of ergothioneine capsules under the current study design.

[0070] In conclusion, after three menstrual cycles of continuous use of ergothioneine capsules, the participants in the trial experienced significant improvements in ovarian reserve, endocrine levels, menopausal symptoms, sleep quality, and menstrual status. All these improvements were statistically significant. p The presence of ergothioneine (<0.05) and no adverse reactions indicates that ergothioneine capsules have a good effect on improving women's ovarian function and related physical conditions.

[0071] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. Application of ergothionein in the preparation of agents to improve ovarian reserve function.

2. The application according to claim 1, characterized in that, The formulation is suitable for individuals with serum anti-Müllerian hormone levels between 1.0 and 3.0 ng / mL.

3. Application of ergothionein in the preparation of agents to improve bone loss caused by ovarian reserve decline.

4. Application of ergothionein in the preparation of formulations to improve menopausal symptoms caused by declining ovarian reserve.

5. The application of ergothioneine in the preparation of hormone-regulating agents, characterized in that, The sex hormones are selected from any one or a combination of at least two of estrogen, follicle-stimulating hormone, luteinizing hormone, or testosterone.

6. Application of ergothionein in the preparation of preparations for regulating menstrual cycle disorders.

7. The application according to any one of claims 1-6, characterized in that, The formulation also contains pharmaceutically acceptable excipients.

8. The application according to claim 7, characterized in that, The pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, solubilizers, osmotic pressure regulators, surfactants, coating materials, colorants, pH adjusters, antioxidants, antibacterial agents, or buffers.

9. The application according to any one of claims 1-6, characterized in that, The dosage forms of the preparations include tablets, granules, capsules, powders, pills, solutions, syrups, emulsions, suspensions, ointments, or microspheres.

Images