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

Abstract

The present application relates to the field of medical technology, and specifically discloses an application of finerenone in the preparation of a drug for treating and / or preventing ovarian dysfunction. Provides a new safe and effective composition for treating ovarian dysfunction. Finerenone, as a main component for preparing a drug for treating and / or preventing ovarian dysfunction, can be administered orally, which reduces the pain and harm of the patient and is more convenient, providing a safe and effective new option for the application of drugs for treating and / or preventing ovarian dysfunction.

Description

Technical Field

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

[0002] In clinical practice, ovarian dysfunction diseases, such as premature ovarian insufficiency (POI) and diminished ovarian reserve (DOR) and women with over-age infertility, all face similar ovarian aging dilemmas, that is, the ovaries lack antral follicles (i.e., larger antral follicles) that can respond well to hormones, making it difficult to obtain high-quality oocytes, so existing assisted reproductive technologies are helpless. Therefore, to find effective treatments for these diseases, we must first start from the root cause and clarify which aspects of the ovarian and follicular development disorders lead to the occurrence of these ovarian diseases. Unfortunately, the diagnosis and treatment methods for ovarian dysfunction diseases are still 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 over-age.

[0003] Among these gynecological diseases, the incidence of premature ovarian failure accounts for 1-4% of fertile women, and with the changes in people's lifestyle, environment, life pressure and childbearing age, it has shown an increasing trend year by year. Premature ovarian failure refers to a clinical syndrome in which women's ovarian physiological activity declines before the age of 40. It is divided into primary and secondary. Regardless of the type of premature ovarian failure, there are symptoms before and after menopause, such as premature aging, osteoporosis, obesity, etc., which make patients age prematurely and bring great pain to their physical and mental health and married life. Premature ovarian failure is clinically manifested as highly heterogeneous and complex causes. To date, although there have been many advances in genetic research on premature ovarian failure, due to the complexity of the causes of premature ovarian failure and the limitations of detection technology, the causes of about 68% of premature ovarian failure are still unclear.

[0004] Inadequate ovarian reserve refers to the process by which the number and quality of follicles in a woman's ovaries gradually decrease. Conditions diagnosed as inadequate ovarian reserve usually include insufficient ovarian reserve markers, poor response to ovarian hormone stimulation, and higher maternal age. Due to the decrease in the number and quality of follicles in patients with inadequate ovarian reserve, the success rate of in vitro fertilization is low in such patients. The incidence of inadequate ovarian reserve is currently increasing year by year, but there is still a lack of effective treatments to improve patients' fertility.

[0005] Although gonadotropin-based IVF has become a programmed assisted reproductive technology to help female patients have children, some patients with poor ovarian response (POR) or POI have only small follicles in their ovaries that do not respond to gonadotropin (IVF). Injection of gonadotropin can not obtain or can only obtain a very small number of antral follicles, which is a huge difference compared to the 10 to 15 antral follicles produced by women with normal ovarian response. As a result, these patients cannot achieve the purpose of reproduction through conventional IVF. Therefore, systematic research on the activation and growth of small follicles is of great significance for the treatment of infertility caused by POR and POI. However, clinical research in the field of primordial follicle activation is almost blank. In 2008, the applicant published an article proposing the hypothesis that PTEN-PI3K controls the activation of small follicles. Based on this theoretical basis, Professor Xue Renwang of Stanford University designed the in vitro activation technology (In vitro activation, IVA) in 2013 to achieve a breakthrough in the treatment of POI infertility. This technology uses laparoscopic surgery to remove part of the ovaries of POI patients and vitrify them for cryopreservation. After the ovarian masses are revived, they are trimmed into smaller cubes to block the Hippo signaling pathway; at the same time, PTEN inhibitors and PI3K agonists are added to the culture to activate the PI3K-Akt signaling pathway. These cultured ovarian masses are then transplanted back into the subfallopian tube membrane of POI patients to wait for the growth and development of follicles. When the follicles in the ovarian masses develop to the antral follicle stage, eggs can be retrieved for conventional in vitro fertilization and assisted reproduction. Through this IVA technology, POI patients can have children. The application of IVA technology in clinical trials has brought hope for fertility in POI patients and provided a new idea for the clinical treatment of POI fertility.

[0006] However, since the PTEN inhibitors and PI3K used in this IVA technology are not FDA-approved safe drugs, there is a high risk of carcinogenesis when these agents are used to treat ovarian tissue. At the same time, this technology lacks consideration of individual differences in different POI patients when used. The treatment method of activating primordial follicles with the same drug is likely to cause negative feedback of PI3K downstream molecules, resulting in unsatisfactory activation of primordial follicles. At present, the success rate of this IVA technology is only about 15%, which is far from meeting clinical needs. More importantly, this IVA technology requires laparoscopic surgery, which undoubtedly brings greater pain and harm to patients. Therefore, oral, intramuscular, intraperitoneal, and ovarian in situ injections may be more convenient for the treatment of ovarian developmental disorders. Therefore, it is particularly important to explore new safe and effective drugs and methods of administration for the treatment of ovarian dysfunction, and the existing technology needs to be improved and developed. 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 drug 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, ovarian hyporesponsiveness, early menopause, ovarian function damage, ovarian insufficiency and ovarian dysfunction.

[0011] The second aspect of the present application provides a 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 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, ovarian hyporesponsiveness, 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, the composition comprising at least one of finerenone, finerenone hydrate, 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, ovarian hyporesponsiveness, 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 prior art, this application has the following advantages:

[0020] The present application provides an application of finerenone in the preparation of a drug for treating and / or preventing 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 with the IVA technology in the prior art that requires laparoscopic surgery, finerenone, as the main ingredient for preparing a drug for treating and / or preventing ovarian dysfunction, can be administered orally, reducing the pain and harm of the patient and being more convenient, providing a safe and effective new option for the application of drugs for treating and / or preventing 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] Figure 1 Experimental flow chart provided for the embodiments of the present application;

[0023] Figure 2 Ovarian tissue diagram provided for the embodiments of the present application;

[0024] Figure 3 This is an ovarian tissue staining analysis diagram provided in the examples of this application;

[0025] Figure 4 A diagram of the number of follicles provided in the embodiments of the present application;

[0026] Figure 5 Western blot analysis diagram provided in the examples of this application;

[0027] Figure 6 Western blot analysis diagram provided in the examples of this application;

[0028] Figure 7 Western blot analysis diagram provided in the examples of this application;

[0029] Figure 8 This is an analysis diagram of mRNA expression provided in the examples of this application;

[0030] Fig. 9 Western blot analysis diagram provided in the examples of this application;

[0031] Fig.10 Experimental flow chart provided for the embodiments of the present application;

[0032] Fig.11 Ovarian tissue diagram provided for the embodiments of the present application;

[0033] Fig.12 Ovarian tissue weight analysis diagram provided for the embodiments of the present application;

[0034] Fig.13 This is an ovarian tissue staining analysis diagram provided in the examples of this application;

[0035] Fig.14 A diagram of the number of follicles provided in the embodiments of the present application;

[0036] Fig.15 Ovarian tissue diagram provided for the embodiments of the present application;

[0037] Fig.16 Ovarian tissue weight analysis diagram provided for the embodiments of the present application;

[0038] Fig.17 This is an ovarian tissue staining analysis diagram provided in the examples of this application;

[0039] Fig.18 A diagram of the number of follicles provided in an embodiment of the present application. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solutions and advantages of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the drawings and embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments of the present application, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the protection scope of the present application. In the absence of conflict, the following embodiments and features in the 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 of "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 technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the 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 the field can implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such 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 mainly includes premature ovarian failure, insufficient ovarian reserve, premature ovarian insufficiency, ovarian hyporesponsiveness, early menopause, ovarian damage, ovarian insufficiency and low ovarian function. Premature ovarian failure (POF) refers to the occurrence of amenorrhea for more than 4 to 6 months before the age of 40, with two tests of follicle-stimulating hormone (FSH) exceeding 40U / L at an interval of more than 4 weeks, accompanied by decreased estrogen and menopausal symptoms. Premature ovarian failure is one of the main causes of female infertility.

[0045] Ovarian reserve refers to the primordial follicles contained in the female ovarian cortex. After birth, the number of primordial follicles will no longer increase, and the number of primordial germ cells in the ovarian cortex will no longer increase. Diminished / decreased ovarian reserve (DOR), also known as low ovarian reserve function, reduced ovarian reserve function, decreased ovarian reserve, decreased ovarian reserve, etc., refers to the number of follicles contained in the ovaries being less than the expected number of follicles, and the ovarian responsiveness and fertility of women of childbearing age being reduced. This can make conception more difficult and reduce the chances of receiving in vitro fertilization (IVF) and other fertility treatments. Compared with the miscarriage rate of women without DOR, patients with DOR can also lead to a higher chance of miscarriage. The number and / or quality of oocytes in the ovaries of patients with DOR is reduced, accompanied by increased FSH levels.

[0046] Premature ovarian insufficiency (POI) is a serious ovarian dysfunction disease, which refers to the loss of ovarian function due to follicle exhaustion in women before the age of 40. The clinical manifestations are amenorrhea for 4 consecutive months, rare or frequent menstruation, and follicle-stimulating hormone (FSH) levels exceeding 25U / L twice with an interval of more than 4 weeks, with or without a fluctuating decrease in 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 the low response of the ovaries to stimulating drugs and the small number of eggs obtained when women undergo controlled superovulation for assisted reproductive technology. Ovarian poor responsiveness is considered an early sign of decreased ovarian reserve.

[0049] 2. Prevention and treatment

[0050] Prevention refers to the preventive treatment of subclinical disease states, which aims to reduce the probability of clinical disease states. Prevention can be divided into primary prevention and secondary prevention. Primary prevention is defined as treatment of subjects who have not yet presented with clinical disease states, while secondary prevention is defined as preventing the secondary occurrence of the same or similar clinical disease states.

[0051] Treatment refers to the treatment 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 composition for treating and / or preventing ovarian dysfunction disease is not a safe drug approved by the FDA and has unsatisfactory effects.

[0053] Based on this, an embodiment of the present application provides a 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, mainly used to treat diabetic nephropathy and heart failure. It is a drug approved by the US Food and Drug Administration (FDA).

[0055] After using finerenone to culture the ovaries of newborn mice in vitro, the ovarian tissue can continue to develop into mature follicles with antrums until ovulation in the subsequent in vivo development. Based on this result, attempts were made to test it through intraovarian injection and oral administration, proving that finerenone can significantly promote the development of secondary follicles and antral follicles in the ovaries of aged mice. This further shows that finerenone can be used as a safe and effective ingredient in drugs for the treatment and / or prevention of ovarian dysfunction diseases.

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

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

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

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

[0060] In one embodiment, the administration of the composition in the examples of the present application includes intraovarian injection and / or oral administration.

[0061] In one embodiment, the composition in the examples of the present application includes a pharmaceutically acceptable auxiliary material, which includes but is not limited to at least one of a pharmaceutical carrier, a diluent, an adjuvant, and an excipient.

[0062] In one embodiment, the composition in the examples of the present application is a tablet, capsule, solution, granule, pill, powder, paste, pill, suspension, powder, injection, suppository, cream or spray. In one embodiment, the composition in the examples of the present application can also be a sustained release preparation, a controlled release preparation or a targeted preparation.

[0063] The following will provide further explanation through a number of specific embodiments.

[0064] Example 1

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

[0066] The specific steps include: separating the paired ovaries, culturing them in serum-free 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 as follows: Figure 1 shown.

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

[0068] Two ovarian tissue blocks from the finerenone group and the control group were dehydrated, embedded, and stained with hematoxylin. Figure 3 As shown. Figure 3It can be seen that after 21 days of implantation, large antral follicles appeared in the finerenone group, while most of the control group were secondary follicles. The statistical analysis of the follicles of all levels in the ovarian tissue of the control group and the finerenone group showed that the following results were obtained: Figure 4 As shown. Figure 4 It can be seen that the number of primary follicles and antral follicles in the mouse ovaries treated with finerenone in vitro in the subsequent in vivo development was significantly increased compared with the control group ovaries, indicating that finerenone treatment in vitro can promote the development of small follicles in mice in vivo.

[0069] Example 2

[0070] The ovaries of 10-day-old mice were digested with collagenase, and the attached granulosa cells were obtained by culturing with serum. The obtained granulosa cells were cultured in serum-free medium for 24 hours to starve the cells, and then treated with finerenone for 15 minutes, 30 minutes, and 60 minutes, respectively. Western blot detection of Akt, 4E-BP1, rs6, and ERK in the ovarian tissues of the three groups of samples and one untreated control sample was performed. The results are shown in Figure 5 to Figure 7 As shown. Figure 5 to Figure 7 It can be seen that the level of phosphorylated ERK (p-ERK) increased significantly 15min, 30min and 60min after finerenone treatment; the phosphorylated Akt (p-Akt) began to increase significantly 15min after finerenone treatment; the phosphorylated 4E-BP1 (p-4EBP1) and phosphorylated rpS6 (p-rpS6) increased 15, 30 and 60min 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 as follows Figure 8 As shown, from Figure 8 It can be seen that finerenone treatment can significantly increase the expression of KITL mRNA.

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

[0073] Western blot was performed to detect the cleaved caspase 3 protein in granulosa cells treated with finerenone and untreated granulosa cells. The results are shown in Fig. 9 As shown. Fig. 9 It can be seen that 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] Select 12-13 month old mice, inject hyaluronic acid into one ovary of the old mice, and inject phenerenone dissolved in hyaluronic acid into the other ovary. The experimental process is as follows: Fig.10 As shown. Ten days later, the mice were dissected and the ovarian tissues were taken for observation. The ovarian tissues of the mice injected with hyaluronic acid alone were the control group, and the ovarian tissues of the mice injected with finerenone dissolved in hyaluronic acid were the finerenone group. The results are shown in Fig.11 and Fig.12 As shown. Fig.11 and Fig.12 It can be seen that the volume and weight of the ovarian tissue in the finerenone group were greater than those in the control group.

[0076] Subsequently, the ovarian tissues of the finerenone group and the control group were dehydrated, embedded, and stained with hematoxylin for analysis. Fig.13 and Fig.14 As shown. Fig.13 and Fig.14 It can be seen that treatment with finerenone can significantly promote the development of primordial follicles and primary follicles in aged mice to higher-order follicles, which provides new evidence for the treatment of ovarian developmental disorders and ovarian aging. In addition, it was found that dissolving finerenone in hyaluronic acid to prepare a solvent that can be injected into the ovaries can provide a basis for the use of finerenone in the preparation of drugs for the treatment and / or prevention of ovarian disorders.

[0077] Example 4

[0078] Old mice aged 12 to 13 months were selected and divided into a control group and a phenerenone group. The mice in the phenerenone group were intragastrically administered with phenerenone (3.2 mg / kg) once a week for three consecutive weeks. The mice in the control group and the phenerenone group were dissected and the ovarian tissues were obtained for observation. Fig.15 and Fig.16 As shown. Fig.15 and Fig.16 It can be seen that the size and weight of the ovarian tissue of the mice in the finerenone group were significantly greater than those of the mice in the control group.

[0079] Subsequently, the ovarian tissues of the finerenone group and the control group were dehydrated, embedded, and stained with hematoxylin for analysis. Fig.17 and Fig.18 As shown. Fig.17 and Fig.18It can be seen that the number of ovarian antral follicles in the finerenone group increased compared with the control group. The statistical counting results showed that there was no significant difference in the number of primordial follicles, primary follicles and larger antral follicles in the ovaries of the control group and the finerenone-treated group. However, the number of secondary follicles in the finerenone ovaries increased significantly compared with the control group, and the number of early antral follicles also increased significantly compared with the control group. It further shows that oral finerenone can significantly promote the development of secondary follicles and early antral follicles in the ovaries of aged mice. It provides a basis 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 and is a drug approved by the U.S. Food and Drug Administration (FDA). Compared with the IVA technology in the prior art that requires laparoscopic surgery, finerenone, as the main ingredient for preparing drugs for treating and / or preventing ovarian dysfunction, can be administered orally, which reduces the pain and harm of patients and is more convenient, providing a safe and effective new option for the preparation of drugs for treating and / or preventing ovarian dysfunction.

[0081] It should be understood that the application of the present application is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made 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 drug for treating and / or preventing diseases related to follicular development disorders; The disease related to follicular development disorder is at least one of insufficient ovarian reserve function, premature ovarian insufficiency, and ovarian hyporesponsiveness.

Images