Use of akkermansia muciniphila in polycystic ovary syndrome products

Abstract

This invention, entitled "Application of *Akkermansia myxophilus* in Products for Polycystic Ovary Syndrome (PCOS)," belongs to the field of microbial technology. The technical problem it aims to solve is the shortcomings of existing PCOS treatment drugs, such as gastrointestinal adverse reactions and unsatisfactory efficacy, and the lack of safe and effective novel intervention strategies. The key technical solution is the application of *Akkermansia myxophilus* (accession number CGMCC No. 20955) in the preparation of drugs for PCOS intervention. The *Akkermansia myxophilus* of this invention can effectively improve clinical signs such as menstrual cycle disorders, obesity, and acne in obese PCOS patients, synergistically improve insulin resistance and abnormal glucose and lipid metabolism, regulate hormone homeostasis, and restore the diversity and structural homeostasis of the intestinal flora. It also exhibits no significant adverse reactions and good safety. This invention provides a novel, safe, and effective intervention strategy for PCOS.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to the application of Akkermansia myxophilus in products for polycystic ovary syndrome. Background Technology

[0002] Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in women of reproductive age, characterized by hyperandrogenemia, ovulation dysfunction, and polycystic ovary morphology. This disease not only significantly increases the risk of infertility but is also associated with an increased incidence of adverse pregnancy outcomes such as miscarriage, preeclampsia, premature birth, and fetal growth restriction, posing a serious threat to women's reproductive health. Notably, obesity and PCOS share the common pathophysiological feature of insulin resistance, which further exacerbates the body's metabolic burden and significantly increases the risk of type 2 diabetes, cardiovascular disease, and metabolic syndrome. Therefore, given the close link between PCOS and metabolic dysfunction, metformin is widely used to treat obese patients with PCOS. However, this drug has gastrointestinal adverse reactions and its efficacy in non-obese patients is poor, highlighting the urgent need to develop safer and more effective treatment options.

[0003] The gut is the largest microbial ecosystem in the human body, and mounting evidence suggests that the gut microbiota plays a crucial role in regulating host energy metabolism, inflammatory responses, and endocrine homeostasis. Furthermore, gut microbiota dysbiosis is closely associated with metabolic diseases such as obesity, diabetes, and polycystic ovary syndrome (PCOS). Studies have found that compared to healthy women, PCOS patients exhibit significantly reduced gut microbial diversity, characterized by increased abundance of potentially pathogenic bacteria such as *Escherichia coli* and *Shigella*, and decreased abundance of beneficial short-chain fatty acid-producing bacteria such as *Ruminaceae*, *Trichophyton*, and *Aktermannii*. Gut microbiota dysbiosis may lead to reduced short-chain fatty acid synthesis, impaired intestinal barrier integrity, and the induction of chronic low-grade inflammation, thereby reducing insulin sensitivity and exacerbating metabolic and endocrine abnormalities associated with PCOS. Against this backdrop, probiotic intervention targeting the gut microbiota has emerged as a novel treatment strategy for PCOS and has attracted widespread attention.

[0004] Relevant patent documents retrieved: The publication, published in China (CN119139358A) on December 17, 2024, discloses a composition for the prevention and treatment of polycystic ovary syndrome (PCOS), comprising the following active ingredients: Tripterygium wilfordii polysaccharide, Dendrobium nobile polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide, and Akkermansia muciniphila suspension. This composition is more effective than oral administration of Akkermansia muciniphila suspension alone.

[0005] Relevant non-patent literature retrieved: The journal or book title is "Frontiers of Medicine," the article title is "Akkermansiamuciniphila PROBIO therapy promotes arginine biosynthesis and reverses reproductive impairments in polycystic ovary syndrome rats," and the publication date is 2025-12-01. This article discloses that Akkermansia muciniphila PROBIO can promote arginine biosynthesis and reverse reproductive impairments in polycystic ovary syndrome rats.

[0006] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects: The strains found in the relevant patent literature have limited regulatory effects on insulin. The *Akkermansia myxophilus* used in the patents has limited regulatory effects on insulin resistance and cannot adequately improve the core metabolic disorders in PCOS patients. The relevant evidence is as follows: based on the patent literature... Figure 5 According to the data, the AKK intervention group reduced fasting blood glucose by only about 48% compared to the model group.

[0007] The strains found in relevant non-patent literature can only modulate part of the gut microbiota structure to a limited extent and cannot effectively correct the abnormal accumulation of pathogenic bacteria and abundance of beneficial bacteria associated with PCOS. The relevant evidence is as follows: based on non-patent literature... Figure 3 According to Chinese patent literature, after intervention with AKK PROBIO, the Shannon index of rats in this group showed no significant difference compared to that in the polycystic ovary syndrome model group; based on non-patent literature... Figure 3 According to the Chinese EF study, after intervention with AKK PROBIO, the levels of Bacteroidetes were reduced compared to the model group, but there was no significant regulatory effect on Proteobacteria, Fusobacteria, and Ruminococci.

[0008] In solving the above problems or overcoming the above defects, the present invention encountered the following difficulties and obstacles: Screening for specific strains from a large number of Akkermansia myxophilus strains that can effectively improve metabolic disorders associated with polycystic ovary syndrome (PCOS) and correct abnormal gut microbiota presents challenges: (1) The strains are highly specific and there are no clear screening criteria. Akkermansia myxophilus strains from different sources and with different preservation numbers have vastly different effects on regulating PCOS-related gut microbiota (such as Proteobacteria, Fusobacteria, and Ruminococci), improving insulin resistance, and reproductive function. Furthermore, there are currently no clear molecular markers to predict the function of the strains, making it impossible to quickly screen them through simple detection. Complete animal experiments and preliminary clinical validation are required for each candidate strain, resulting in a huge workload for screening. (2) There are many screening indicators with complex correlations. Effective strains must simultaneously meet multiple indicators such as "significantly improving insulin resistance, correcting the enrichment of pathogenic bacteria and abnormal abundance of beneficial bacteria, and regulating hormone homeostasis." There are mutual influences among the indicators, and some strains may only improve a single indicator, making it difficult to achieve multi-dimensional intervention effects at the same time, which increases the difficulty and complexity of screening. Summary of the Invention

[0009] The purpose of this invention is to provide: The application of Akkermansia myxophilus in products for polycystic ovary syndrome (PCOS), and related technologies, to address technical issues such as gastrointestinal adverse reactions and poor efficacy of PCOS treatment drugs, as well as the lack of safe and effective new intervention options, or combinations thereof.

[0010] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0011] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0012] The definition of standard chemical terminology can be found in the reference "Microbiology", edited by Shen Ping and Chen Xiangdong, Higher Education Press.

[0013] Unless otherwise specified, conventional methods within the scope of the art, such as culture medium preparation, strain culture, bacterial dilution, and colony counting, shall be used.

[0014] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0015] The terms "optional" or "arbitrarily" mean that the event or situation subsequently described may or may not occur, including both the occurrence and non-occurrence of the event or situation. For example, according to the definition below: the inactivated bacteria include any one or more of pasteurized bacteria, moist heat-inactivated bacteria, and dry heat-inactivated bacteria. Examples: the inactivated bacteria may include pasteurized bacteria, or the inactivated bacteria may include moist heat-inactivated bacteria, or the inactivated bacteria may include dry heat-inactivated bacteria, or the inactivated bacteria may include pasteurized bacteria, moist heat-inactivated bacteria, and dry heat-inactivated bacteria.

[0016] The term "Akkermansia muciniphila" used in this article refers to Akkermansia muciniphila with accession number CGMCC No.20955, which can be prepared into live bacteria, inactivated bacteria, bacterial powder, metabolites and other forms through high-density fermentation, inactivation and other processes. It has biological activities that regulate intestinal flora homeostasis and improve insulin resistance.

[0017] The term “polycystic ovary syndrome (PCOS)” used in this article refers to an endocrine and metabolic disorder in women of reproductive age characterized by hyperandrogenemia, ovulation dysfunction, and polycystic ovary morphology. It is often accompanied by complications such as insulin resistance, gut microbiota dysbiosis, and obesity, and includes two subtypes: obese and non-obese.

[0018] The term "intestinal flora homeostasis" used in this article refers to the state in which the relative abundance of various flora in the gut (including Bacteroidetes, Proteobacteria, Fusobacteria, etc.) is in balance, the ratio of beneficial bacteria (such as Akkermania and Faecalibacterium) to potentially pathogenic bacteria (such as some Fusobacterium and Ruminococcus) is appropriate, the diversity and community structure of the intestinal flora are normal, and the host's energy metabolism, inflammatory response and endocrine homeostasis can be maintained.

[0019] The term "insulin resistance" used in this article refers to the decreased sensitivity of the body's tissue cells to insulin, which prevents insulin from effectively promoting glucose uptake and utilization, resulting in elevated fasting blood glucose and fasting insulin levels. It is a core pathophysiological feature shared by PCOS and obesity.

[0020] The term "postgenetic agent" used in this article refers to the products of Akkermansia myxophilus that retain their biological activity after being processed by inactivation (such as pasteurization), fermentation, extraction, etc., including inactivated bacteria, bacterial extracts, fermentation supernatants, freeze-dried bacterial preparations, etc., which have the advantages of high stability, good safety and ease of formulation development.

[0021] In a first aspect, the present invention provides: Akkermansia myxophila ( Akkermansia muciniphila Its application in the preparation of drugs for the intervention of polycystic ovary syndrome.

[0022] The technical features include: Akkermansia myxophilus, pharmaceuticals, and polycystic ovary syndrome.

[0023] Specifically, the accession number of the *Ackermania* strain is CGMCC No. 20955.

[0024] Specifically, the drug comprises any one or more of the following: culture of Akkermansia myxophilus, inactivated bacteria, bacterial powder, metabolites, bacterial extract, fermentation supernatant, and lyophilized bacterial preparation.

[0025] Preferably, the inactivated bacteria include any one or more of pasteurized bacteria, moist heat inactivated bacteria, and dry heat inactivated bacteria.

[0026] More preferably, the pasteurization conditions are 80-90℃ for 10-30 minutes.

[0027] More preferably, the pasteurization inactivation conditions are 80-81, 81-82, 82-83, 83-84, 84-85, 85-86, 86-87, 87-88, 88-89 or 89-90 min for inactivation of 10-11, 11-12, 12-13, 13-14, 14-15, 15-16, 16-17, 17-18, 18-19, 19-20, 20-21, 21-22, 22-23, 23-24, 24-25, 25-26, 26-27, 27-28, 28-29 or 29-30 min.

[0028] Specifically, the drug contains at least 1×10 6 CFU / dose of Ackermansia myxophila.

[0029] Preferably, the drug contains at least 1×10 6 -1×10 12 CFU / dose of Ackermansia myxophila.

[0030] More preferably, the drug contains at least 1×10 6 -1×10 7 1×107 1×10 8 1×10 8 -1×10 9 1×10 9 -1×10 10 1×10 10 -1×10 11 Or 1×10 11 -1×10 12 CFU / dose of Ackermansia myxophila.

[0031] More preferably, the drug contains at least 1×10 9 -1×10 10 Or 1×10 10 -1×10 11 CFU / dose of Ackermansia myxophila.

[0032] Specifically, the medicine also includes pharmaceutically acceptable excipients or nutritional additives.

[0033] Preferably, the excipients include any one or more of the following: binders, fillers, disintegrants, lubricants, preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators.

[0034] Preferably, the nutritional additives include one or more of dietary fiber, prebiotics, protein, lipids, minerals, and vitamins.

[0035] In some specific embodiments of the present invention, the excipient is maltodextrin. These specific embodiments are merely examples and are not intended to limit the scope of protection of the present invention. Any product containing *Akkermansia muciniphila* (accession number CGMCC No. 20955) falls within the scope of protection of the present invention.

[0036] Specifically, the dosage form of the medicine includes any one or more of capsules, tablets, granules, powders, oral liquids, suspensions, emulsions, and injections.

[0037] Specifically, the polycystic ovary syndrome includes any one or more of obese polycystic ovary syndrome and non-obese polycystic ovary syndrome.

[0038] Specifically, the drug is used to improve the clinical signs of patients with obesity-related polycystic ovary syndrome, improve insulin resistance and glucose and lipid metabolism disorders, regulate hormone homeostasis, and restore intestinal flora homeostasis.

[0039] Preferably, the improvement in clinical signs includes reduction in weight, hip circumference and acne score, and improvement in menstrual cycle disorders, hirsutism and acanthosis nigricans; Preferably, the improvement of insulin resistance and glucose and lipid metabolism disorders includes reducing fasting insulin levels, insulin resistance index, and low-density lipoprotein cholesterol levels.

[0040] Preferably, the hormone homeostasis regulation includes reducing luteinizing hormone levels, the luteinizing hormone / follicle-stimulating hormone ratio, and testosterone levels; Preferably, the restoration of gut microbiota homeostasis includes increasing gut microbiota α diversity, decreasing the relative abundance of Proteobacteria, Fusobacterium, Bacteroides and Fusobacterium, and increasing the relative abundance of Actinobacteria, Bifidobacterium and Faecalibacterium.

[0041] The present invention has at least the following beneficial effects: Compared with existing technologies, the present invention has better technical effects in regulating insulin resistance and correcting PCOS-related pathogenic bacteria enrichment and abnormal beneficial bacteria abundance.

[0042] According to experimental tests, the present invention improves the effect of reducing fasting blood glucose from 48% compared to the model group in the prior art to 66.17% compared to the model group.

[0043] According to experimental tests, this invention reduced the relative abundance of Fusobacteria from no significant change in the prior art to a significant decrease compared to the model group (p<0.05), reduced the relative abundance of Rumenococci from no significant change in the prior art to a significant decrease compared to the model group (p<0.01), and reversed the decrease in the relative abundance of Bacteroidetes from the prior art to a significant increase compared to the model group, thus more comprehensively correcting PCOS-related dysbiosis.

[0044] Considering the possibility of this invention entering other countries, this invention also provides the following technical solutions: This invention provides a method for preventing or treating polycystic ovary syndrome, the method comprising using Akkermansia myxophilus with accession number CGMCC No. 20955 or a product containing Akkermansia myxophilus with accession number CGMCC No. 20955.

[0045] Specifically, the method includes administering an effective amount of Akkermansia myxophilus to the subject.

[0046] Preferably, the subjects include mammals.

[0047] More preferably, the subject is a human being.

[0048] Specifically, the effective amount is ≥1×10 6 CFU / dose of Ackermansia myxophila.

[0049] Preferably, the effective amount is 1×106 -1×10 12 CFU / dose of Ackermansia myxophila.

[0050] More preferably, the effective amount is 1×10⁻⁶. 6 -1×10 7 1×10 7 1×10 8 1×10 8 -1×10 9 1×10 9 -1×10 10 1×10 10 -1×10 11 Or 1×10 11 -1×10 12 CFU / dose of Ackermansia myxophila.

[0051] More preferably, the effective amount is 11 × 10⁻⁶. 9 -1×10 10 Or 1×10 10 -1×10 11 CFU / dose of Ackermansia myxophila.

[0052] Preservation Instructions Preserved strain: AKK PROBIO; Category Naming: Akkermansia muciniphila ; Accession number: CGMCC No. 20955; Date of preservation: October 26, 2020; Preservation institution: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Culture Collections; Abbreviation of depositary institution: CGMCC; Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Attached Figure Description

[0053] Figure 1 Flowchart for subject recruitment and randomization.

[0054] Figure 2The study considered changes in clinical signs in patients with polycystic ovary syndrome (PCOS); A. weight changes; B. body mass index changes; C. hip circumference changes; D. acne scores in the two groups after intervention; E. changes in acne scores. Groups: P (placebo group, n=45); AKK (Aklerella myxophilus post-biotic intervention group, n=47); data are expressed as mean ± standard deviation (SD) or median ± interquartile range (IQR); the Shapiro-Wilk test was used to verify the normality of the dataset; statistical comparisons between groups were performed using independent samples t-test or Mann-Whitney U test, depending on the normality of the data distribution; the significance level was set at *. p <0.05,** p <0.01.

[0055] Figure 3 This study analyzed changes in insulin resistance and lipid metabolism indicators, including: A. Changes in fasting blood glucose levels; B. Changes in fasting insulin levels; C. Changes in the insulin resistance index; and D. Changes in low-density lipoprotein cholesterol levels. Groups: P (placebo group, n=45); AKK (Akkermansia myxophilus postbiotic intervention group, n=47). Data are expressed as mean ± standard deviation (SD) or median ± interquartile range (IQR). The Shapiro-Wilk test was used to verify the normality of the dataset. Statistical comparisons between groups were performed using the independent samples t-test or the Mann-Whitney U test, depending on the normality of the data distribution. The significance level was set at *. p <0.05,** p <0.01.

[0056] Figure 4 The analysis focused on changes in hormone levels. A. Comparison of luteinizing hormone (LH) levels between the two groups after the intervention; B. Changes in LH levels; C. Changes in the LH / Follicle-stimulating hormone (FSH) ratio; D. Changes in testosterone concentration. Groups: P (placebo group, n=45); AKK (Aklerella myxotropicis post-biotic intervention group, n=47). Data are expressed as mean ± standard deviation (SD) or median ± interquartile range (IQR). The Shapiro-Wilk test was used to verify the normality of the dataset. Statistical comparisons between groups were performed using either the Independent Samples t-test or the Mann-Whitney U test, depending on the normality of the data distribution. The significance level was set at *. p <0.05.

[0057] Figure 5Differences in gut microbiota structure among groups were analyzed using: A) Gut microbiota α diversity index analysis (A) Shannon index, B) Allen H index, C) Rao quadratic entropy index, and D) Faith phylogenetic diversity index; E) Principal coordinate analysis; and F) Linear discriminant analysis (effect size score > 3.06, used to screen for differentially expressed microbiota between groups). Grouping: HC (healthy control group, n=30); P (placebo group, n=30); AKK (Ackermannia myxophilus postbiotic intervention group, n=30). Data are expressed as mean ± standard deviation (SD) or median ± interquartile range (IQR). The Shapiro-Wilk test was used to verify the normality of the dataset. One-way repeated-measures ANOVA was used for comparisons between groups of continuous variables, and Tukey's test or Dunn's test was used for correction of multiple comparisons. The significance level was set at *. p <0.05,** p <0.01, *** p <0.001.

[0058] Figure 6 The composition and differences of gut microbiota at the phylum and genus levels were analyzed for each group. A. Phylum-level species distribution characteristics; B. Relative abundance comparison of Actinobacteria, Proteobacteria, and Fusobacterium; E. Genus-level species distribution characteristics; F. Relative abundance comparison of Bifidobacterium, Bacteroides, Faecalibacterium, Ruminococcus, Fusobacterium, and Akkermansia; L. Random forest analysis (used to screen for differentially expressed marker microbiota between groups). Groups: HC (healthy control group, n = 30); P (placebo group, n = 30); AKK (Akkermansia myxophilus postbiotic intervention group, n = 30). Data are expressed as mean ± standard deviation or median ± interquartile range. The Shapiro-Wilk test was used to verify the normality of the dataset. One-way repeated-measures ANOVA was used for comparisons between groups of continuous variables, and Tukey's test or Dunn's test was used for correction of multiple comparisons. The significance level was set at *. p <0.05,** p <0.01, *** p <0.001. Detailed Implementation

[0059] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0060] The present invention will be further described below by way of specific embodiments. All instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention, unless otherwise specified, are obtained through conventional commercial channels. The *Akermansia muciniphila* used in the present invention (… Akkermansia muciniphila, The AKK (abbreviated as AKK) has accession number CGMCC No.20955 and has been disclosed in patent CN116925975B.

[0061] Example 1: Research Design and Research Subjects 1. Research subjects and inclusion / exclusion criteria This randomized, double-blind, placebo-controlled clinical trial was conducted at Ganzhou People's Hospital from November 2024 to August 2025. The study protocol was approved by the Ethics Committee of Ganzhou People's Hospital (Approval No.: TY-ZKY2024-090-01) and registered with the Chinese Clinical Trial Registry (Registration No.: ChiCTR2400090949). All study procedures followed the ethical principles of the Declaration of Helsinki and were strictly implemented in accordance with the 2025 CONSORT reporting guidelines.

[0062] The study participants were women with obesity-related polycystic ovary syndrome. All participants were fully informed about the study content and signed a written informed consent form before enrollment.

[0063] Inclusion criteria: (1) diagnosed with polycystic ovary syndrome according to the Rotterdam criteria; (2) having insulin resistance (homeostatic model insulin resistance index ≥2.69) and obesity (body mass index ≥28 kg / m²). 2 (3) Age 18-40 years old; (4) No use of drugs that affect menstruation in the past 3 months and no related treatment; (5) No organic lesions of the reproductive tract or other primary diseases; (6) Willing to comply with the research protocol.

[0064] Exclusion criteria: (1) Comorbid endocrine diseases (such as diabetes mellitus, thyroid dysfunction); (2) Severe systemic diseases (liver, kidney, cardiovascular, autoimmune diseases or mental illness); (3) History of thromboembolism; (4) Smoking or alcohol abuse; (5) Pregnancy or lactation; (6) Malignant tumors; (7) Use of probiotics, antibiotics or statins in the past month; (8) History of drug allergy; (9) Congenital malformation of the reproductive tract.

[0065] Withdrawal criteria: (1) the occurrence of intolerable adverse reactions; (2) the occurrence of a medical emergency requiring alternative treatment; (3) the subject voluntarily withdraws; (4) the subject fails to cooperate with the study according to the study protocol.

[0066] 2. Basic Information of the Research Object of this Invention To evaluate the effect of *Akermansia myxophilus* postbiotics on clinical symptoms in obese women with polycystic ovary syndrome (PCOS), this invention conducted a prospective randomized controlled trial. A total of 128 PCOS patients were initially screened, of whom 28 were excluded due to not meeting the inclusion criteria: 16 refused to participate, 4 did not meet the inclusion criteria due to recent antibiotic use, and 8 withdrew for other reasons.

[0067] This invention ultimately enrolled 100 participants, who were randomly assigned in a 1:1 ratio to the placebo group (P group) and the *Akkermansia myxophilus* postbiotic group (AKK group), with 50 participants in each group. During the follow-up period, 92 participants completed the trial (45 in the placebo group and 47 in the *Akkermansia myxophilus* postbiotic group), while the remaining participants were lost to follow-up or voluntarily discontinued the intervention (see...). Figure 1 ).

[0068] There were no statistically significant differences in baseline characteristics between the two groups, indicating good balance. Specific data are as follows: the mean age in the placebo group (P group) was 28.22 ± 3.83 years, and in the Akkermansia myxophilus postbiotic intervention group (AKK group) was 28.21 ± 3.98 years; there was no statistically significant difference between the groups. p =0.990); the median body mass index in the placebo group was 29.04 (28.28, 30.68) kg / m². 2 The post-biotic group of *Ackermania pseudomallei* was 29.69 (28.13, 31.61) kg / m³. 2 All met the inclusion criteria for obese polycystic ovary syndrome, and there were no statistically significant differences between the groups. p =0.419); the median insulin resistance index in the placebo group was 4.81 (3.12, 6.81), and in the Akkermansia myxophilus post-biotic group it was 5.19 (3.20, 6.57), both meeting the inclusion criteria for insulin resistance, with no statistically significant difference between the groups. p =0.751). In addition, there were no statistically significant differences between the two groups in terms of menstrual regularity, waist-to-hip ratio (WHR), hirsutism score, sex hormone levels (follicle-stimulating hormone, luteinizing hormone, testosterone, etc.) and other key baseline indicators (see Table 1), which further suggests that the baseline balance is good and the interference of baseline confounding factors on the intervention effect can be excluded.

[0069] Table 1. Baseline data of patients upon enrollment

[0070] Note: Menstrual regularity is defined as a categorical variable (regular: menstrual cycle 21-35 days; irregular: menstrual cycle <21 days or >35 days). Groups: P group (placebo group, n=45); AKK group (Akkermansia myxophilus postbiotic intervention group, n=47). Data are expressed as mean ± standard deviation or median (interquartile range). Independent samples t-tests were used for comparisons between groups of normally distributed continuous data; Mann-Whitney U tests were used for non-normally distributed continuous data; chi-square (χ²) tests were used for categorical data.

[0071] Example 2: Intervention Program and Measurement Indicators 1. Drug preparation The Akkermansia myxophilus (AKK) used in this study was isolated from healthy Chinese individuals and provided by Suncon Biotechnology (Suzhou) Co., Ltd. Each capsule contains 1×10⁻⁶ spores. 10 Pasteurized and inactivated *Ackermania myxophilus* cells. The preparation process of the postbiotic formulation was as follows: *Ackermania myxophilus* with preservation number CGMCC No. 20955 was fermented at high density, then heat-inactivated at 80±5℃ for 30 min (ensuring complete inactivation of the cells while retaining bioactive components), homogenized with maltodextrin, and finally filled into hydroxypropyl methylcellulose empty capsules (Zhejiang Xinchang Kangping Capsule Co., Ltd.). The placebo capsules were filled with an equal amount of xylooligosaccharides in the same capsules. Clinical evaluations were conducted at baseline (week 0) and at the end of the intervention (week 12).

[0072] The placebo capsules were identical in appearance to the trial capsules. Independent researchers, unaware of the group assignments, repackaged the intervention drug and placebo capsules, labeling each capsule with a unique participant code. The trial was double-blind throughout, and participants, researchers, and outcome assessors were unaware of the group assignments until data analysis was completed.

[0073] 2. Dosing regimen Both groups of subjects received standard treatment for polycystic ovary syndrome (oral contraceptives, referring to PCOS treatment guidelines). Dosing began on day 5 of the menstrual cycle, one tablet daily for 21 consecutive days per cycle, with a total treatment cycle of 3 menstrual cycles (12 weeks in total).

[0074] In addition to the standard treatment, the placebo group (P group) took one placebo capsule orally daily for 12 weeks; the Akkermansia myxotropic metabiotic group (AKK group) took one capsule containing Akkermansia myxotropic metabiotic orally daily for 12 weeks.

[0075] 3. Biochemical indicator testing Fasting venous blood was collected from subjects at baseline and at week 12. After serum separation, the following indicators were measured: (1) Insulin resistance-related indicators: fasting blood glucose, fasting insulin, and homeostatic model insulin resistance index (calculated using the homeostatic model method). (2) Glucose and lipid metabolism indicators: glycated hemoglobin, triglycerides, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol; (3) Sex hormone indicators: follicle-stimulating hormone, luteinizing hormone, and testosterone; (4) Liver and kidney function related indicators: alanine aminotransferase, aspartate aminotransferase, creatinine, uric acid.

[0076] All biochemical indicators were tested using standardized laboratory methods and a fully automated biochemical analyzer.

[0077] 4. 16S rRNA high-throughput sequencing After the intervention, fecal samples were collected from the subjects. Total genomic DNA was extracted from the samples using the Tiangen Biotech Fecal DNA Extraction Kit (DP328). The DNA concentration and purity were detected using a Thermo Fisher Scientific NanoDrop spectrophotometer. Qualified DNA samples were sent to Shanghai Paisenno Biotechnology Co., Ltd. for further analysis. The V4 region of the 16S rRNA gene was amplified by PCR, followed by paired-end sequencing using the Illumina HiSeq2000 platform. Sequencing data were analyzed using QIIME2 (version 2019.4). After quality filtering and chimera removal, clusters were established based on 97% sequence similarity as operational taxonomic units. Alpha diversity (Shannon index) and β diversity (based on Bray-Curtis distance principal coordinate analysis) were calculated to assess the gut microbiota structure.

[0078] 5. Statistical Analysis All statistical analyses were performed using GraphPad Prism 9.0 software (GraphPad Software, San Diego, California, USA). Data distribution was assessed using the Shapiro-Wilk test. Based on the normality of the data distribution, data are expressed as mean ± standard deviation (SD) or median (interquartile range, IQR). Independent samples t-tests or Mann-Whitney U tests were used for comparisons between two groups; one-way repeated measures ANOVA was used for comparisons of three or more groups, followed by post-hoc multiple comparisons using the Tukey test or Dunn test. The χ² test was used for comparisons of categorical variables. In two-tailed tests, a p-value <0.05 was considered statistically significant.

[0079] Example 3: Acetobacter xanthophyte postbiotics improves clinical signs in obese patients with polycystic ovary syndrome. Studies have shown that obese patients with polycystic ovary syndrome (PCOS) often present with menstrual cycle disorders, increased waist-to-hip ratio, and clinical signs such as hirsutism, acne, and acanthosis nigricans. After the intervention, a 3-month follow-up showed that menstrual cycles improved in both groups, with the AKK group showing better results: the proportion of patients with regular menstrual cycles in the placebo group (P group) increased from 24.44% to 40.00%, and in the Akkermansia myxophilus post-proliferator group, it increased from 19.15% to 40.43% (see Table 2). After the intervention, the weight of the AKK group was slightly lower than that of the P group, with no statistically significant difference between the groups, but the weight loss in the AKK group was significantly greater than that in the P group (P group vs. AKK group = 0.37±1.54 vs. 1.92±2.62). p <0.001, Figure 2 (A in the original text); Meanwhile, the decrease in body mass index in the AKK group was significantly greater than that in the P group (P group vs. AKK group = 0.15±0.62 vs. 0.75±1.06). p <0.01, Figure 2 (B in the table). In addition, waist circumference, hip circumference and waist-to-hip ratio of both groups decreased compared with before the intervention, and the improvement was more obvious in the AKK group (see Table 1 and Table 2); among them, the hip circumference reduction of the AKK group was significantly greater than that of the P group (P group vs. AKK group = 2.00 (0.00, 4.00) vs. 3.00 (2.00, 5.00)). p <0.05, Figure 2 The C in the figure suggests that AKK post-biotics can significantly reduce the subjects' weight and hip circumference. Regarding clinical signs, both groups showed improvement in hirsutism, acne, and acanthosis nigricans after intervention, but the improvement was more pronounced in the AKK group (see Tables 1 and 2). Compared with the placebo group, the acne score was significantly lower in the Akkermansia myxophilus post-biotic group after intervention (P group vs. AKK group = 1.00 (0.50, 2.00) vs. 1.00 (0.00, 1.00)). p <0.01, Figure 2 In the D group, the acne score decreased more significantly (P group vs. AKK group = 1.00 (0.50, 2.00) vs. 2.00 (1.00, 2.00)). p <0.05, Figure 2 The results (E) indicate that the intervention of *Akermansia myxophilus* metabiotics can improve signs such as hirsutism, acne, and acanthosis nigricans, with the most significant effect on acne.

[0080] In conclusion, combining conventional treatment with Akkermansia myxophilus postbiotic can more effectively improve menstrual cycle disorders, obesity, hirsutism, acne, acanthosis nigricans, and other clinical signs in obese polycystic ovary syndrome patients.

[0081] Table 2. Patient indicators after intervention

[0082] Note: Menstrual regularity is defined as a categorical variable (regular: menstrual cycle 21-35 days; irregular: menstrual cycle <21 days or >35 days). Groups: P group (placebo group, n=45); AKK group (Akkermansia myxophilus postbiotic intervention group, n=47). Data are expressed as mean ± standard deviation or median (interquartile range, IQR). Independent samples t-tests were used for comparisons between groups of normally distributed continuous data; Mann-Whitney U tests were used for non-normally distributed continuous data; chi-square (χ²) tests were used for categorical data.

[0083] Example 4: Synergistic effect of Acetobacter myxotrophic metabolites on insulin resistance and glucose and lipid metabolism Obese patients with polycystic ovary syndrome (PCOS) often have concurrent glucose and lipid metabolism disorders and insulin resistance, which significantly increase their risk of type 2 diabetes and cardiovascular disease. After 12 weeks of intervention, all metabolic indicators in both groups improved to varying degrees, with the *Aklerotica myxotropicis* postbiotic intervention group showing significantly better improvement than the placebo group (see Table 2). Assessment of insulin resistance showed that fasting blood glucose, fasting insulin levels, and insulin resistance index decreased in both groups after intervention, with a more pronounced reduction in the AKK group. Compared to the placebo group, the AKK group showed a slightly greater reduction in fasting blood glucose (…). Figure 3 (A) but there was no statistically significant difference between the groups; while the AKK group had a significantly higher reduction in fasting insulin than the placebo group (P group vs. AKK group = 3.73 (2.51, 6.58) vs. 6.61 (4.16, 11.09). p <0.01, Figure 3 (B in the text). In addition, the reduction in insulin resistance index in the AKK group was significantly greater than that in the placebo group (P group vs. AKK group = 1.25 (0.69, 2.02) vs. 1.75 (1.19, 3.03). p <0.05, Figure 3 (C) The above results suggest that *Akkermansia myxophilus* metabolites can effectively improve insulin resistance in obese patients with PCOS. Glycolipid metabolism profile analysis showed that after the intervention, glycated hemoglobin levels in both groups decreased, with the AKK group showing a better reduction than the placebo group, but the difference between the groups was not statistically significant (see Tables 1-2). Regarding blood lipids, total cholesterol levels in the AKK group decreased significantly compared to before the intervention; triglyceride and high-density lipoprotein cholesterol levels changed, but the differences between the groups were not significant; while the reduction in low-density lipoprotein cholesterol in the AKK group was significantly greater than that in the placebo group (P group vs. AKK group = 0.10±0.32 vs. 0.23±0.42). p <0.01, Figure 3 (D in the middle).

[0084] In summary, the metabiotics of Akkermansia myxophilus have a synergistic effect with conventional treatment, which can further improve the metabolic homeostasis of obese patients with polycystic ovary syndrome, providing new ideas and experimental evidence for the clinical intervention of this disease.

[0085] Example 5: Acetaminophen regulates hormone homeostasis in Akkermansia myxophilus with good safety profile. Elevated luteinizing hormone (LH) / follicle-stimulating hormone (FSH) ratio and hyperandrogenemia are important pathophysiological features of obesity-related polycystic ovary syndrome (PCOS). After 12 weeks of intervention, both groups showed significantly reduced LH / FSH ratios and testosterone levels (Tables 1-2). After intervention, there was no statistically significant difference in FSH levels between the two groups, but the LH level in the Akkermansia myxotropic bacteria post-biotic intervention group was significantly lower than that in the placebo group (P<0.05 vs. AKK group = 5.89 ± 3.49 vs. 4.67 ± 2.05). p <0.05, Figure 4 The A group showed a significantly greater decrease in luteinizing hormone (LH) levels than the placebo group (P group vs. AKK group = 1.98 ± 3.32 vs. 3.67 ± 3.84). p <0.05, Figure 4 (B in the original text). Meanwhile, the decrease in the luteinizing hormone / follicle-stimulating hormone ratio in the Akkermansia myxophilus post-biotic intervention group was significantly greater than that in the placebo group (P group vs. AKK group = 0.37±0.67 vs. 0.70±0.73). p <0.05, Figure 4 (C in the text). Furthermore, the effect of Akkermansia myxophilus metabiotics on reducing testosterone levels was significantly better than the placebo group (P group vs. AKK group = 4.97±6.20 vs. 7.74±6.87). p <0.05, Figure 4 (D in the middle).

[0086] To further verify the biosafety of Akkermansia myxophilus postbiotic, multiple liver and kidney function-related indicators of the subjects were tested. The results showed that the levels of alanine aminotransferase, aspartate aminotransferase, creatinine and uric acid in the Akkermansia myxophilus postbiotic intervention group remained stable before and after the intervention (Table 2). All safety indicators remained within the normal reference range, and no intervention-related adverse reactions occurred during the study.

[0087] In summary, the metabiotic of Akkermansia myxophilus can effectively regulate hormonal homeostasis in obese patients with polycystic ovary syndrome and has good biocompatibility.

[0088] Example 6: Ackermania myxophilus postbiotics restore the diversity and structure of the gut microbiota To evaluate the impact of *Akermansia myxophilus* postbiotics on the gut microbiota of obese patients with polycystic ovary syndrome (PCOS), this study performed 16S rRNA high-throughput sequencing analysis on fecal samples collected after intervention. Age-matched healthy women were included as a healthy control group (HC group), and gut microbiota composition and diversity were compared and analyzed simultaneously. α-diversity analysis showed that the Shannon index and Faith phylogenetic diversity index of the gut microbiota in the P group were lower than those in the HC group, with the Allen H index (…) being significantly lower. p <0.001) and Rao's second entropy exponent ( p The difference between the P group and the HC group was <0.05, indicating a significant decrease in the species richness and genetic diversity of the gut microbiota in obese polycystic ovary syndrome patients. Furthermore, intervention with *Akermansia myxophilus* postbiotics significantly reversed the decreasing trend of gut microbiota α diversity in the P group (see [link to relevant documentation]). Figure 5 The AD in the model effectively increases the species richness and genetic diversity of the gut microbiota. Principal coordinate analysis showed that the gut microbiota structure of the HC group and the P group exhibited a clear clustering and separation trend, indicating that there are differences in the community structure of the gut microbiota between the two groups (see [link to analysis]). Figure 5 (E in the text); After intervention with Akkermansia myxophila postbiotics, the gut microbiota structure of the AKK group was more similar to that of the HC group, suggesting that this intervention can effectively improve the gut microbiota dysbiosis in obese polycystic ovary syndrome patients (see E in the text). Figure 5 The results of linear discriminant analysis further confirmed that multiple differentially enriched bacterial genera were present in the HC, P, and AKK groups, and the gut microbiota composition characteristics of each group showed significant differences (see E in the original text). Figure 5 (F in the middle).

[0089] In summary, Akkermansia myxophilus postbiotics can effectively restore the homeostasis of gut microbiota structure in obese polycystic ovary syndrome patients by increasing the species richness and genetic diversity of gut microbiota.

[0090] Example 7: The post-biotic regulation of gut microbiota composition in obese patients with polycystic ovary syndrome by *Akermansia myxophilus* To further elucidate the impact of *Akermansia myxophila* postbiotics on the gut microbiota composition of obese patients with polycystic ovary syndrome (PCOS), this study performed phylum and genus-level species composition analysis on sequencing data. Phylum-level analysis showed that the gut microbiota of both healthy women and obese PCOS patients were dominated by Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria (see [link to study].) Figure 6 In group A); compared with group HC, group P had Proteobacteria ( p <0.001) and Fusobacteria ( p The relative abundance of <0.05 was significantly increased, while the abundance of Actinobacteria (p The relative abundance was significantly reduced to <0.001 (see [reference needed]). Figure 6 BD in the group); while the AKK group of Proteobacteria ( p =0.17) and Fusobacteria ( p <0.01) relative abundance was lower than group P, Actinobacteria ( p =0.13) The relative abundance was higher than that of group P (see Figure 6 The study found that *Akermansia myxophilus* postbiotics could reverse the increase in abundance of Proteobacteria and Fusobacteria and the decrease in abundance of Actinobacteria in the gut of obese patients with polycystic ovary syndrome (PCOS). Genus-level analysis showed significant differences in the genus-level composition of the gut microbiota among the three groups (see [link to study]. Figure 6 In the E group); among them, the beneficial bacteria in group P are Bifidobacterium and Faecalibacterium ( p <0.001) and Akkermania ( p The relative abundance of <0.001) was lower than that of the HC group, while the opportunistic pathogens Bacteroides and Ruminococcus were also present. p <0.05) and the relative abundance of Fusobacterium was higher than that of the HC group ( Figure 6 The FK group). Compared with group P, the postbiotic intervention of *Ackermania myxophilus* increased the relative abundance of *Bifidobacterium* and *Femtobacter*, while decreasing the abundance of *Bacteroides* and *Ruminococcus*. p <0.01) and Fusobacterium ( p The relative abundance of <0.05) decreased, thus making the gut microbiota composition of obese polycystic ovary syndrome patients closer to the level of healthy individuals. Figure 6 The FJ group in the study). Random forest analysis further confirmed the presence of multiple differentially expressed bacterial genera among the groups, with the AKK group showing a similar trend to the HC group (see FJ). Figure 6 L in the middle.

[0091] In summary, Akkermansia myxophilus postbiotics can regulate gut microbiota composition by increasing gut microbiota diversity, enriching beneficial bacteria, and reducing potentially harmful bacteria, thereby restoring gut microbiota homeostasis in obese polycystic ovary syndrome patients and providing a key microecological regulatory basis for improving their clinical symptoms and metabolic indicators.

[0092] Experimental Example 8: Akkermansia myxophilus improves polycystic ovary syndrome in female rats Fifteen 6-week-old SPF-grade female SD rats were randomly divided into three groups of five each: a normal control group (Control group), a model control group (Model group), and an Akkermansia muciniphila intervention group (AKK group). After one week of acclimatization, the rats were used for experiments.

[0093] Rats in the Control group were administered an equal volume of 0.5% sodium carboxymethyl cellulose (CMC-Na) by gavage at a dose of 10 mL / kg / day for 28 consecutive days. Rats in the other two groups were administered an equal volume of letrozole by gavage and fed an equal amount of high-sugar, high-fat diet at a dose of 1 mg / kg / day for 28 consecutive days to induce a PCOS rat model. All three groups of rats were housed at room temperature of 22±2℃, relative humidity of 55±5%, and with a light / dark cycle of 12h / 12h.

[0094] After modeling, rats in both the Control and Model groups were treated with 10 mL / kg / day of distilled water by gavage once daily for 21 days; AKK rats were treated with 1×10 9 CFU / mL / kg / d of Ackermania myxophilus postbiotic (Ackermania myxophilus with preservation number CGMCC No.20955, after high-density fermentation, was heat-inactivated at 85±5℃ for 30 min), once daily, continuously administered by gavage in equal volume for 21 days.

[0095] One day before the end of the gavage treatment, rats in each group were fasted for 12 hours but allowed free access to water. Blood was then collected by tail clipping, and the fasting insulin levels of the rats in each group were measured using an ELISA kit via enzyme-linked immunosorbent assay (ELISA). The results are shown in Table 3. Table 3. Fasting insulin levels in rats of each group

[0096] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. Akkermansia myxophila ( Akkermansia muciniphila Its application in the preparation of drugs for the intervention of polycystic ovary syndrome is characterized by, The preservation number of the *Ackermania* strain is CGMCC No. 20955.

2. The application according to claim 1, characterized in that, The drug comprises any one or more of the following: culture of Akkermansia myxophilus, inactivated bacteria, bacterial powder, metabolites, bacterial extract, fermentation supernatant, and lyophilized bacterial preparation.

3. The application according to claim 2, characterized in that, The inactivated bacteria include any one or more of pasteurized bacteria, moist heat inactivated bacteria, and dry heat inactivated bacteria.

4. The application according to claim 1, characterized in that, The drug contains at least 1×10 6 CFU / dose of Ackermansia muciniphila.

5. The application according to claim 4, characterized in that, The medicine contains 1×10 6 -1×10 12 CFU / dose of Ackermansia muciniphila.

6. The application according to claim 1, characterized in that, The medicines also include pharmaceutically acceptable excipients or nutritional additives.

7. The application according to claim 6, characterized in that, The excipients include any one or more of the following: binders, fillers, disintegrants, lubricants, preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators.

8. The application according to claim 6, characterized in that, The nutritional additives include one or more of the following: dietary fiber, prebiotics, protein, lipids, minerals, and vitamins.

9. The application according to claim 1, characterized in that, The dosage forms of the medicine include any one or more of capsules, tablets, granules, powders, oral liquids, suspensions, emulsions, and injections.

10. The application according to claim 1, characterized in that, The polycystic ovary syndrome mentioned above includes any one or more of obese polycystic ovary syndrome and non-obese polycystic ovary syndrome.

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