Use of finerenone in preparation of drug for treating and / or preventing ovarian dysfunction diseases

Abstract

The present invention relates to the technical field of pharmaceuticals. Particularly, disclosed is use of finerenone in the preparation of a drug for treating and / or preventing ovarian dysfunction diseases. Provided is a novel, safe, and effective composition for treating ovarian dysfunction diseases. Finerenone, as a main component in the preparation of the drug for treating and / or preventing the ovarian dysfunction diseases, can be applied orally, which reduces the pain and injury to patients and is more convenient, thereby providing a safe and effective new choice for the use in the preparation of the drug for treating and / or preventing the ovarian dysfunction diseases.

Description

Application of finerenone in preparing medicine for treating and / or preventing ovarian dysfunction Technical Field

[0001] The present application relates to the field of medical technology, and in particular to a use of finerenone in the preparation of a drug for treating and / or preventing ovarian dysfunction. Background Art

[0002] Currently, the primary challenge facing my country's population development is low fertility. Over the past five years, the number of newborns has declined by approximately 40%. Despite the relaxation of the "three-child policy," this declining fertility trend has been unable to reverse. By 2022, only 9 million newborns will be born nationwide. Female fertility is the cornerstone of increasing the population's fertility rate. Infertility affects approximately 15% of couples of childbearing age. Clinically, ovarian dysfunction, such as premature ovarian insufficiency (POI), diminished ovarian reserve (DOR), and post-fertility age, all face the same dilemma of ovarian aging: a lack of hormonally responsive antral follicles (i.e., large, antral follicles), making it difficult to obtain high-quality oocytes. Consequently, existing assisted reproductive technologies are unable to address this problem. Therefore, to find effective treatments for these conditions, we must first address the root causes and identify the underlying impairments in ovarian and follicular development that contribute to these ovarian diseases. Unfortunately, diagnostic and treatment options for ovarian dysfunction remain very limited. There is no effective way to reverse the above-mentioned ovarian diseases, such as premature ovarian failure, decreased ovarian reserve and infertility caused by excessive age.

[0003] Among these gynecological conditions, premature ovarian failure (POF) occurs in 1-4% of women of childbearing age. With changes in lifestyle, environment, stress, and reproductive age, the incidence of POF is increasing annually. POF refers to a clinical syndrome characterized by a decline in ovarian activity in women before the age of 40. It can be categorized as either primary or secondary. Regardless of the type, POF presents with premenopausal symptoms, such as premature aging, osteoporosis, and obesity, leading to premature aging and significant distress to both physical and mental health and sexual well-being. POF is clinically highly heterogeneous and has complex etiologies. Despite significant progress in genetic research, the etiology of approximately 68% of POF remains unclear due to its complex etiology and limitations in detection technology.

[0004] Deficient ovarian reserve (DOR) refers to a gradual decrease in the number and quality of follicles in a woman's ovaries. Conditions typically diagnosed as DOR include insufficient ovarian reserve markers, a poor response to ovarian hormone stimulation, and advanced maternal age. Due to the reduced number and quality of follicles in patients with DOR, in vitro fertilization (IVF) success rates are low. The incidence of DOR is increasing annually, yet effective treatments to improve fertility remain lacking.

[0005] Although gonadotropin-based IVF has become a standard assisted reproductive technology for women with fertility, some patients with poor ovarian response (POR) or POI (Poor Ovarian Response Intrauterine Insemination) (POI) have only small follicles in their ovaries that are unresponsive to gonadotropins (IVF). Gonadotropin injections can produce no or very few antral follicles, a significant difference from the 10 to 15 antral follicles produced by women with normal ovarian response. Consequently, these patients are unable to achieve fertility through conventional IVF. Therefore, systematic research on the activation and growth of small follicles is crucial for treating infertility caused by POR and POI. However, clinical research in the field of primordial follicle activation is virtually nonexistent. In 2008, the applicant published a paper proposing the hypothesis that PTEN-PI3K controls follicle activation. Based on this hypothesis, Professor Xue Renwang of Stanford University designed an in vitro follicle activation (IVA) technique in 2013, achieving a breakthrough in treating POI infertility. This technique uses laparoscopic surgery to remove portions of ovaries from POI patients and vitrify them for cryopreservation. After thawing, the ovarian masses are trimmed into smaller cubes to block the Hippo signaling pathway. Simultaneously, a PTEN inhibitor and PI3K agonist are added to the culture to activate the PI3K-Akt pathway. These cultured ovarian masses are then transplanted back into the POI patient's subfallopian tube to await follicle development. Once the follicles in the ovarian masses develop to the antral follicle stage, eggs can be retrieved for conventional in vitro fertilization (IVF). This IVA technique has enabled POI patients to become fertile. The application of IVA in clinical trials offers hope for fertility in POI patients and provides a novel approach to clinical treatment.

[0006] However, because the PTEN inhibitors and PI3K inhibitors used in this IVA technique are not FDA-approved, treating ovarian tissue with these agents carries a high risk of carcinogenesis. Furthermore, this technique lacks consideration for the individual differences among POI patients. Using the same drug to activate primordial follicles is likely to result in negative feedback loops on downstream PI3K molecules, leading to suboptimal activation of primordial follicles. Currently, the success rate of this IVA technique is only approximately 15%, far from meeting clinical needs. More importantly, this IVA technique requires laparoscopic surgery, which undoubtedly causes greater pain and harm to patients. Therefore, oral, intramuscular, intraperitoneal, and in situ ovarian injections may be more convenient treatments for ovarian development disorders. Therefore, the exploration of new, safe, and effective drugs and delivery methods for treating ovarian dysfunction is crucial, and existing technologies require further improvement and development. Summary of the Invention

[0007] In view of the above-mentioned deficiencies in the prior art, the purpose of the present application is to provide a use of finerenone in the treatment and / or prevention of ovarian dysfunction diseases.

[0008] The technical solution of this application is as follows:

[0009] In a first aspect of the present application, there is provided a use of finerenone in the preparation of a medicament for treating and / or preventing ovarian dysfunction.

[0010] Optionally, the ovarian dysfunction disease includes at least one of premature ovarian failure, insufficient ovarian reserve function, premature ovarian insufficiency, poor ovarian response, early menopause, ovarian function damage, ovarian insufficiency and ovarian dysfunction.

[0011] In a second aspect of the present application, a method for preparing a drug for treating and / or preventing ovarian dysfunction is provided, wherein the method comprises: providing a hydrate of finerenone, a solvate of finerenone, a pharmaceutically acceptable salt of finerenone, or a polymorph of finerenone for use in preparing a drug for treating and / or preventing ovarian dysfunction.

[0012] Optionally, the ovarian dysfunction disease includes at least one of premature ovarian failure, insufficient ovarian reserve function, premature ovarian insufficiency, poor ovarian response, early menopause, ovarian function damage, ovarian insufficiency and ovarian dysfunction.

[0013] The third aspect of the present application provides a composition for treating and / or preventing ovarian dysfunction, wherein the composition comprises at least one of finerenone, a finerenone hydrate, a finerenone solvate, a pharmaceutically acceptable salt of finerenone, and a finerenone polymorph.

[0014] Optionally, the ovarian dysfunction disease includes at least one of premature ovarian failure, insufficient ovarian reserve function, premature ovarian insufficiency, poor ovarian response, early menopause, ovarian function damage, ovarian insufficiency and ovarian dysfunction.

[0015] Optionally, the composition is administered by intraovarian injection and / or oral administration.

[0016] Optionally, the composition includes a pharmaceutically acceptable excipient.

[0017] Optionally, the auxiliary material is hyaluronic acid.

[0018] Optionally, the composition is in the form of tablets, capsules, solutions, granules, pills, powders, ointments, pills, suspensions, powders, injections, suppositories, creams or sprays.

[0019] Compared with the existing technology, this application has the following advantages:

[0020] The present application provides the use of finerenone in the preparation of a drug for the treatment and / or prevention of ovarian dysfunction. Finerenone is an oral selective nonsteroidal mineralocorticoid receptor antagonist and is a drug approved by the U.S. Food and Drug Administration (FDA). Compared to the prior art IVA technology that requires laparoscopic surgery, finerenone, as the main ingredient in the preparation of a drug for the treatment and / or prevention of ovarian dysfunction, can be administered orally, reducing pain and harm to patients and providing greater convenience. This provides a safe and effective new option for the preparation of drugs for the treatment and / or prevention of ovarian dysfunction. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for describing the embodiments.

[0022] FIG1 is a flow chart of an experiment provided in an embodiment of the present application;

[0023] FIG2 is a diagram of ovarian tissue provided in an embodiment of the present application;

[0024] FIG3 is a staining analysis diagram of ovarian tissue provided in an embodiment of the present application;

[0025] FIG4 is a diagram showing the number of follicles provided in an embodiment of the present application;

[0026] FIG5 is a Western blot analysis diagram provided in an embodiment of the present application;

[0027] FIG6 is a Western blot analysis diagram provided in an embodiment of the present application;

[0028] FIG7 is a Western blot analysis diagram provided in an embodiment of the present application;

[0029] FIG8 is an analysis diagram of mRNA expression levels provided in the examples of the present application;

[0030] FIG9 is a Western blot analysis diagram provided in an embodiment of the present application;

[0031] FIG10 is a flowchart of an experiment provided in an embodiment of the present application;

[0032] FIG11 is a diagram of ovarian tissue provided in an embodiment of the present application;

[0033] FIG12 is a graph showing ovarian tissue weight analysis provided in an embodiment of the present application;

[0034] FIG13 is a staining analysis diagram of ovarian tissue provided in an embodiment of the present application;

[0035] FIG14 is a diagram showing the number of follicles provided in an embodiment of the present application;

[0036] FIG15 is a diagram of ovarian tissue provided in an embodiment of the present application;

[0037] FIG16 is a graph showing ovarian tissue weight analysis provided in an embodiment of the present application;

[0038] FIG17 is a staining analysis diagram of ovarian tissue provided in an embodiment of the present application;

[0039] FIG18 is a diagram showing the number of follicles provided in an embodiment of the present application. DETAILED DESCRIPTION

[0040] To make the purpose, technical solutions and advantages of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below in conjunction with the drawings and embodiments of this application. Obviously, the described embodiments are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application. In the absence of conflict, the features in the following embodiments and embodiments can be combined with each other.

[0041] It should be noted that if there are descriptions involving "first", "second", etc. in the implementation of this application, the descriptions such as "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or suggesting their relative importance and implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of such features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the fact that ordinary technicians in this field can implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0042] To facilitate understanding of the embodiments of the present application, several terms involved in the present application are briefly introduced below.

[0043] 1. Ovarian dysfunction

[0044] Ovarian dysfunction primarily includes premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, poor ovarian response, premature menopause, ovarian damage, ovarian insufficiency, and low ovarian function. Premature ovarian failure (POF) refers to amenorrhea for more than four to six months before the age of 40, with two follicle-stimulating hormone (FSH) levels exceeding 40 U / L measured more than four weeks apart, accompanied by decreased estrogen levels and menopausal symptoms. Premature ovarian failure is one of the main causes of female infertility.

[0045] Ovarian reserve refers to the number of primordial follicles within a woman's ovarian cortex. After birth, the number of primordial follicles and the number of primordial germ cells within the ovarian cortex no longer increase. Diminished / decreased ovarian reserve (DOR), also known as impaired ovarian reserve, decreased ovarian reserve, decreased ovarian reserve, or reduced ovarian reserve, refers to a condition in which the ovaries contain fewer follicles than expected, leading to decreased ovarian responsiveness and fertility in women of childbearing age. This can make conception more difficult and reduce access to in vitro fertilization (IVF) and other fertility treatments. Compared to women without DOR, women with DOR also have a higher risk of miscarriage. Women with DOR experience a decrease in the number and / or quality of oocytes in their ovaries, accompanied by elevated FSH levels.

[0046] Premature ovarian insufficiency (POI) is a serious ovarian dysfunction disorder that occurs before the age of 40 due to follicular depletion. Clinical manifestations include amenorrhea, oligomenorrhea, or frequent menstruation for four consecutive months, follicle-stimulating hormone (FSH) levels exceeding 25 U / L twice at intervals of more than four weeks, and with or without fluctuating estrogen levels.

[0047] Premature menopause refers to entering menopause before the normal age of menopause, before the age of 45.

[0048] Poor ovarian response (POR), also known as poor ovarian response, refers to a low ovarian response to stimulating drugs and a low number of eggs retrieved during controlled ovarian hyperstimulation for assisted reproductive technology. Poor ovarian response is considered an early sign of diminished ovarian reserve.

[0049] 2. Prevention and treatment

[0050] Prevention refers to the preventive treatment of subclinical disease states, aiming to reduce the probability of clinical disease states developing. Prevention can be categorized as primary prevention and secondary prevention. Primary prevention is defined as treatment of subjects who have not yet developed a clinical disease state, while secondary prevention is defined as preventing a secondary occurrence of the same or similar clinical disease state.

[0051] Treatment refers to the management of a disease, symptom, or condition, including inhibiting the development of the disease, symptom, or condition and / or delaying or alleviating the disease, symptom, or condition.

[0052] In the prior art, the compositions for treating and / or preventing ovarian dysfunction diseases are not safe drugs approved by the FDA and their effects are not ideal.

[0053] Based on this, the present invention provides an embodiment of the invention for use of finerenone in the preparation of a drug for treating and / or preventing ovarian dysfunction.

[0054] Finerenone is an oral, selective nonsteroidal mineralocorticoid antagonist (MRA) developed by Bayer Healthcare Pharmaceuticals for the treatment of diabetic nephropathy and heart failure. It is a drug approved by the U.S. Food and Drug Administration (FDA).

[0055] After culturing the ovaries of newborn mice with finerenone in vitro, the ovarian tissue continued to develop into mature antral follicles and ovulate in vivo. Based on this result, further testing using intraovarian injection and oral administration demonstrated that finerenone significantly promoted the development of secondary and antral follicles in the ovaries of aged mice. This suggests that finerenone could be a safe and effective ingredient in medications for the treatment and / or prevention of ovarian dysfunction.

[0056] In one embodiment, the ovarian dysfunction disease includes at least one of premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, poor ovarian response, early menopause, ovarian damage, ovarian insufficiency and ovarian hypofunction.

[0057] A second aspect of the present application provides a use of a hydrate, solvate, pharmaceutically acceptable salt, or polymorph of finerenone in preparing a drug for treating and / or preventing ovarian dysfunction. It is understood that the hydrate, solvate, pharmaceutically acceptable salt, or polymorph of finerenone has the same or similar effects as finerenone. Therefore, the hydrate, solvate, pharmaceutically acceptable salt, or polymorph of finerenone can be used in preparing a drug for treating and / or preventing ovarian dysfunction.

[0058] In one embodiment, pharmaceutically acceptable salts include salts formed from alkali metal salts, alkaline earth metal salts and suitable organic ligands.

[0059] In a third aspect of the embodiments of the present application, a composition for treating and / or preventing ovarian dysfunction is provided, comprising at least one of finerenone, a finerenone hydrate, a finerenone solvate, a pharmaceutically acceptable salt of finerenone, and a finerenone polymorph.

[0060] In one embodiment, the composition of the present invention is administered by intraovarian injection and / or oral administration.

[0061] In one embodiment, the composition of the present invention includes pharmaceutically acceptable excipients, including but not limited to at least one of a pharmaceutical carrier, a diluent, an adjuvant, and an excipient.

[0062] In one embodiment, the composition of the present invention is a tablet, capsule, solution, granule, pill, powder, ointment, pill, suspension, powder, injection, suppository, cream or spray. In one embodiment, the composition of the present invention can also be a sustained-release formulation, a controlled-release formulation or a targeted formulation.

[0063] The following will further illustrate this through a number of specific embodiments.

[0064] Example 1

[0065] The ovaries of 7-day-old newborn mice treated with finerenone were implanted under the renal capsule of mice, allowing the ovarian tissue to grow in the mice.

[0066] The specific steps include: isolating paired ovaries, culturing them in serum-free culture medium for 3 days, selecting one of the ovarian tissues, adding finerenone (20 μM) and culturing for 24 hours, and then transplanting the two ovaries of the control group and the finerenone-treated group under the renal capsule of the same recipient mouse. The operation process is shown in Figure 1.

[0067] After 21 days of implantation, the transplanted ovaries were taken for observation, and the results are shown in Figure 2. As can be seen from Figure 2, after 21 days of implantation, the size of the ovarian tissue mass in the finerenone group was significantly increased compared to that in the control group.

[0068] Two ovarian tissue specimens from the finerenone group and the control group were dehydrated, embedded, and stained with hematoxylin for analysis. The results are shown in Figure 3. As shown in Figure 3, 21 days after implantation, large antral follicles appeared in the finerenone group, while the majority of follicles in the control group were secondary follicles. Statistical analysis of follicles of all grades in the ovarian tissues of the control and finerenone groups is shown in Figure 4. As shown in Figure 4, the number of primary and antral follicles in the ovaries of mice treated with finerenone in vitro significantly increased during subsequent in vivo development compared to the ovaries of the control group, indicating that finerenone treatment in vitro can promote the development of small follicles in mice in vivo.

[0069] Example 2

[0070] Ovaries of 10-day-old mice were digested with collagenase, and adherent granulosa cells were obtained by serum culture. Granulosa cells were starved by culturing in serum-free medium for 24 hours and then treated with finerenone for 15, 30, and 60 minutes. Western blot analysis of Akt, 4E-BP1, rs6, and ERK in ovarian tissue from the three groups and one untreated control was performed. The results are shown in Figures 5 to 7. Figures 5 to 7 show that phosphorylated ERK (p-ERK) levels increased significantly after 15, 30, and 60 minutes of finerenone treatment; phosphorylated Akt (p-Akt) levels began to increase significantly 15 minutes after finerenone treatment; and phosphorylated 4E-BP1 (p-4EBP1) and phosphorylated rpS6 (p-rpS6) levels increased 15, 30, and 60 minutes after finerenone treatment.

[0071] The KITL protein downstream of the mTORC1 signaling pathway in granulosa cells was detected, and the expression level of KITL protein was analyzed by detecting the expression level of KITL mRNA. The results are shown in Figure 8. As can be seen from Figure 8, finerenone treatment can significantly increase the expression of KITL mRNA.

[0072] The results in combination with Figures 5 to 8 indicate that finerenone can activate the ERK signaling pathway, PI3K signaling pathway and mTORC1 signaling pathway in mouse granulosa cells.

[0073] Western blot analysis was performed on cleaved caspase 3 protein in granulosa cells treated with finerenone and untreated granulosa cells, and the results are shown in Figure 9. As shown in Figure 9, the expression level of cleaved caspase 3 protein in granulosa cells treated with finerenone was significantly reduced, indicating that finerenone inhibited the apoptosis of ovarian cells.

[0074] Example 3

[0075] Twelve- to thirteen-month-old mice were selected and injected with hyaluronic acid into one ovary and finerenone dissolved in hyaluronic acid into the other. The experimental process is shown in Figure 10. Ten days later, the mice were dissected and ovarian tissue was obtained for observation. Ovarian tissue from mice injected with only hyaluronic acid served as the control group, while ovarian tissue from mice injected with finerenone dissolved in hyaluronic acid served as the finerenone group. The results are shown in Figures 11 and 12. As can be seen from Figures 11 and 12, the volume and weight of the ovarian tissue in the finerenone group were greater than those in the control group.

[0076] Ovarian tissues from the finerenone and control groups were subsequently dehydrated, embedded, and stained with hematoxylin. The results are shown in Figures 13 and 14. As shown in Figures 13 and 14, treatment with finerenone significantly promotes the development of primordial and primary follicles into higher-order follicles in aged mice, providing new evidence for the treatment of ovarian developmental disorders and ovarian aging. Furthermore, the discovery that finerenone can be dissolved in hyaluronic acid to prepare a solution suitable for injection into the ovaries provides a basis for its use in the preparation of drugs for the treatment and / or prevention of ovarian disorders.

[0077] Example 4

[0078] Mice aged 12 to 13 months were divided into a control group and a pheneron group. The mice in the pheneron group were gavaged with 3.2 mg / kg of pheneron weekly for three consecutive weeks. Ovarian tissue was then harvested and observed after dissection. The results are shown in Figures 15 and 16 . As shown in Figures 15 and 16 , the ovarian tissue of the pheneron-treated mice was significantly larger and heavier than that of the control group.

[0079] Ovarian tissues from the finerenone and control groups were subsequently dehydrated, embedded, and stained with hematoxylin for analysis. The results are shown in Figures 17 and 18. As shown in Figures 17 and 18, the number of antral follicles in the finerenone group increased compared to the control group. Statistical counts showed no significant difference in the number of primordial follicles, primary follicles, and larger antral follicles in the ovaries of the control and finerenone-treated groups. However, the number of secondary follicles and early antral follicles in the finerenone-treated ovaries was significantly increased compared to the control group. This further demonstrates that oral administration of finerenone can significantly promote the development of secondary and early antral follicles in the ovaries of aged mice. This provides evidence for the use of finerenone in the preparation of drugs for the treatment and / or prevention of ovarian disorders.

[0080] In summary, finerenone is an oral, selective nonsteroidal mineralocorticoid receptor antagonist approved by the U.S. Food and Drug Administration (FDA). Compared to existing IVA techniques that require laparoscopic surgery, finerenone, as the main ingredient in the preparation of drugs for the treatment and / or prevention of ovarian dysfunction, can be administered orally, reducing pain and harm to patients and providing greater convenience. This provides a safe and effective new option for the preparation of drugs for the treatment and / or prevention of ovarian dysfunction.

[0081] It should be understood that the application of this application is not limited to the above examples. For ordinary technicians in this field, they can make improvements or changes based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to this application.

Claims

1. Use of finerenone in the preparation of a medicament for treating and / or preventing ovarian dysfunction diseases.

2. The application according to claim 1, characterized in that The ovarian dysfunction diseases include at least one of premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, poor ovarian response, premature menopause, ovarian function impairment, ovarian hypofunction, and ovarian hypofunction.

3. Use of a hydrate of finerenone, a solvate of finerenone, a pharmaceutically acceptable salt of finerenone, or a polymorph of finerenone in the preparation of a medicament for treating and / or preventing ovarian dysfunction.

4. The application according to claim 3, characterized in that, The ovarian dysfunction diseases include at least one of premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, poor ovarian response, premature menopause, ovarian function impairment, ovarian hypofunction, and ovarian hypofunction.

5. A composition for treating and / or preventing ovarian dysfunction diseases, characterized in that, The composition includes at least one of finerenone, a hydrate of finerenone, a solvate of finerenone, a pharmaceutically acceptable salt of finerenone, and a polymorph of finerenone.

6. The composition according to claim 5, wherein The ovarian dysfunction diseases include at least one of premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, poor ovarian response, premature menopause, ovarian function impairment, ovarian hypofunction, and ovarian hypofunction.

7. The composition according to claim 5, wherein The administration method of the composition includes intraovarian injection and / or oral administration.

8. The composition according to claim 5, wherein The composition includes pharmaceutically acceptable excipients.

9. The composition according to claim 8, wherein The excipient is hyaluronic acid.

10. The composition according to claim 5, characterized in that, The composition is a tablet, capsule, solution, granule, pill, powder, plaster, bolus, suspension, powder, injection, suppository, cream, or spray.

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