Application of senkyunolide I in preparation of medicine for improving ovarian function decline and / or ovarian insufficiency

Abstract

The invention discloses application of senkyunolide I in preparation of a medicine for improving ovarian function decline and / or ovarian insufficiency, and relates to the technical field of biological medicine. Research finds that senkyunolide I has a remarkable technical effect in the aspect of improving ovarian insufficiency and ovarian function decline. The compound can effectively promote ovarian angiogenesis, provides a good microenvironment for follicle development, and has an obvious regulation effect on morphological development of follicles at all levels. The senkyunolide I can also regulate the mood cycle of disorder, so that the mood cycle tends to be normalized. Besides, the invention also finds that the senkyunolide I not only can improve the ovarian reserve function, but also plays an important role in balance regulation of sex hormone level, and further improves the health state of the reproductive system. Meanwhile, the senkyunolide I has an improvement effect on endometrial thickness and gland development, and also provides structural and functional support for embryo implantation and pregnancy maintenance.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of ligustrolide I in the preparation of drugs for improving ovarian dysfunction and / or ovarian insufficiency. Background Technology

[0002] Decreased ovarian reserve (DOR) refers to a gradual reduction in the number and quality of eggs in a woman's ovaries. This is a natural physiological process that typically occurs as women age. Decreased ovarian reserve is one of the manifestations of a woman's gradually declining fertility. Key characteristics and effects include: reduced egg quantity, decreased egg quality, irregular menstrual cycles, decreased or increased menstrual flow, and decreased fertility, potentially leading to infertility or a longer time required to conceive.

[0003] Long-term deficiency of ovarian endocrine hormones caused by ovarian insufficiency (POI) is a significant pathogenic factor for female infertility and osteoporosis. POI refers to a weakened ability of the ovaries to produce eggs, a decline in follicle quality and quantity, a decrease in oocyte quality, infrequent menstruation, and reduced menstrual flow. It is characterized by elevated serum follicle-stimulating hormone (FSH) levels and decreased serum estradiol (E2) levels, leading to decreased female fertility and reproductive endocrine disorders. Recurrent POI can further develop into premature ovarian failure (POF), a primary ovarian defect characterized by amenorrhea, decreased estrogen levels, and elevated serum gonadotropin levels before the age of 40. POF not only affects the fertility of women of reproductive age but also causes bone loss and severe osteoporosis due to decreased estrogen levels. Its estimated incidence is 1 / 10,000 before age 20, 1 / 1,000 at age 30, and 1 / 100 at age 40. The causes of ovarian infertility (POI) are currently summarized as follows: genetic mutations, autoimmune ovarian damage, iatrogenic factors, the effects of radiotherapy and chemotherapy, and viral infections. Hormone replacement therapy is only used to control disease progression; there is currently no feasible cure. Hormone replacement therapy also significantly increases the incidence of estrogen-dependent tumors such as ovarian cancer and breast cancer, posing a great threat to women's health. Therefore, the main treatment for POI is currently hormone replacement therapy, with traditional Chinese medicine or prepared Chinese medicines used as adjunctive therapies to protect remaining oocytes and gradually restore ovarian function.

[0004] Traditional Chinese medicine (TCM) believes that ovarian dysfunction is caused by various congenital and acquired factors leading to "insufficient Yin water stored in the kidneys, lack of nourishment from the Chong and Ren meridians, insufficient source for the generation of reproductive essence, dysfunction of the ovaries' storage and release functions, resulting in difficulty in egg development and maturation, and imbalance in the uterus's ability to hold substances, making conception difficult." One key aspect is ovarian microcirculatory disorders, which can lead to prolonged ischemia and hypoxia in the ovaries, increasing follicular atresia and causing reproductive endocrine dysfunction. To address this issue, TCM health preservation strategies can regulate the function of ovarian vascular endothelial cells (ECs) through different metabolic pathways, promote ovarian cell proliferation, respond to the stress of the ovarian microenvironment, and further reduce vascular endothelial cell damage. This effectively triggers ovarian angiogenesis and remodeling, providing the blood, nutrients, and hormones necessary for follicle development. The ovary is the most angiogenic reproductive organ; its microcirculatory disorders can lead to ovarian ischemia and hypoxia, accelerating cell apoptosis and even follicular atresia, resulting in ovarian dysfunction. Moreover, the ovary is the only normal tissue in the human body, besides the endometrium, that can undergo cyclical angiogenesis and remodeling. This invention aims to discover natural compounds from traditional Chinese medicinal materials that can improve ovarian dysfunction and ovarian insufficiency, thereby providing technical support for the development of drugs to improve ovarian dysfunction and ovarian insufficiency. Summary of the Invention

[0005] The purpose of this invention is to provide the application of ligustrazine I in the preparation of drugs for improving ovarian dysfunction and / or ovarian insufficiency, thereby addressing the problems existing in the prior art. This invention has found that ligustrazine I has an ameliorative effect on ovarian insufficiency and ovarian dysfunction, thus providing a scientific basis for the development of drugs for improving ovarian dysfunction and ovarian insufficiency.

[0006] To achieve the above objectives, the present invention provides the following solution:

[0007] This invention provides the use of ligustilide I in the preparation of a drug for improving ovarian dysfunction and / or ovarian insufficiency.

[0008] Furthermore, the drug improves decreased ovarian function and / or ovarian insufficiency by improving ovarian angiogenesis.

[0009] Furthermore, the drug improves decreased ovarian function and / or ovarian insufficiency by improving the developmental morphology of follicles at all stages.

[0010] Furthermore, the drug improves decreased ovarian function and / or ovarian insufficiency by increasing ovarian reserve.

[0011] Furthermore, the drug improves decreased ovarian function and / or ovarian insufficiency by regulating sex hormone levels.

[0012] Furthermore, the drug improves decreased ovarian function and / or ovarian insufficiency by improving endometrial thickness and glandular development.

[0013] The present invention also provides a medicament for improving ovarian dysfunction and / or ovarian insufficiency, the active ingredient of which includes ligustrazine I.

[0014] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0015] Furthermore, the excipients include fillers, binders, disintegrants, emulsifiers, flavoring agents, preservatives, or colorants.

[0016] Furthermore, the dosage form of the drug is tablets, granules, capsules, pills, or oral liquid.

[0017] The present invention discloses the following technical effects:

[0018] This invention, through systematic experimental research, has discovered that ligustrolactone I has significant technical effects in improving ovarian insufficiency and decreased ovarian function. This compound effectively promotes ovarian angiogenesis, providing a favorable microenvironment for follicle development, and significantly regulates the morphological development of follicles at all stages, helping to maintain normal ovarian physiological function. Ligustrolactone I can also regulate disordered estrous cycles, normalizing them and thus improving the overall homeostasis of the reproductive endocrine system. Furthermore, this invention has found that ligustrolactone I not only improves ovarian reserve but also plays an important role in the balanced regulation of sex hormone levels, further enhancing the health of the reproductive system. Simultaneously, the improving effect of ligustrolactone I on endometrial thickness and glandular development provides structural and functional support for embryo implantation and pregnancy maintenance. Based on these findings, this invention not only reveals the potential application value of ligustrolactone I in the treatment of ovarian-related diseases but also provides a solid scientific basis and technical support for its development as a core component of novel drugs. These active molecules derived from natural plants have advantages such as high safety and good bioavailability, laying the foundation for subsequent precision treatment strategies targeting ovarian dysfunction and possessing significant clinical translational potential and social application prospects. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a schematic diagram of the technical route of the present invention;

[0021] Figure 2 Microscopic images of the cellular composition of rat vaginal secretions at different time points;

[0022] Figure 3 A statistical graph showing the estrous cycle of rats in each group;

[0023] Figure 4 HE staining images of ovaries of rats in each group (A), and statistical graphs of total number of follicles (B) and number of atretic follicles (C);

[0024] Figure 5 The graph shows the results of serum FSH(A), E2(B), AMH(C), LH(D), and HSD17h3(E) levels in rats of each group after 21 days of treatment.

[0025] Figure 6 HE staining images of the uterus of rats in each group (A) and statistical graph of endometrial thickness (B);

[0026] Figure 7 A schematic diagram of the birth of rats in each group (A) and a statistical graph of the number of offspring obtained by single births (B). Detailed Implementation

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0029] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0030] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0031] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0032] The ligustilide I referred to in this invention has CAS number 94596-28-8 and molecular formula C. 12 H 16 O4, the structural formula is as follows:

[0033]

[0034] Example 1

[0035] 1. Experimental Methods

[0036] (1) Ninety-six healthy, sexually mature female SD rats, aged 8 weeks and weighing 220-250g, were selected and managed under normal conditions with free access to water and food. They were acclimatized for three days before the experiment. The 96 rats were randomly divided into groups using a random number table method. Specific grouping and experimental procedures are as follows (…). Figure 1 )as follows:

[0037] ① Normal control group: equal volume of corn oil was injected intraperitoneally for 28 days;

[0038] ② Ovarian insufficiency model group (Model): A rat model of POI was established by daily intraperitoneal injection of vinyl cyclohexene dioxide (VCD) (80mg / kg / d). The dosage was adjusted according to the weight of each rat. The drug was administered for 15 consecutive days, followed by intraperitoneal injection of an equal amount of corn oil for 28 days.

[0039] ③ Low-dose group of ligustrazine lactone I (SenI L): VCD (80mg / kg / d) was injected intraperitoneally at regular intervals every day. The dosage was adjusted according to the weight of each rat. The treatment was continued for 15 days. Then, ligustrazine lactone I (36mg / kg) dissolved in corn oil was injected intraperitoneally once every other day for a total of 28 days.

[0040] ④ High-dose group of ligustrazine lactone I (SenI H): VCD (80mg / kg / d) was injected intraperitoneally at regular intervals every day. The dosage was adjusted according to the weight of each rat. The treatment was continued for 15 days. Then, ligustrazine lactone I (72mg / kg) dissolved in corn oil was injected intraperitoneally once every other day for a total of 28 days.

[0041] ⑤ Ferrostatin-1 positive control group (Fer-1): VCD (80mg / kg / d) was injected intraperitoneally daily at regular intervals. The dosage was adjusted according to the weight of each rat. The drug was administered for 15 consecutive days. After that, Ferrostatin-1 (Fer-1) (2.5mg / kg), a ferroptosis inhibitor dissolved in corn oil, was injected intraperitoneally every other day for a total of 28 days.

[0042] (2) Results monitoring and data collection

[0043] Every two days, the weight, food intake, and estrous cycle changes of rats in each group were measured and recorded.

[0044] On day 28 after drug administration, tissue samples were collected, and morphological changes in the ovaries were observed using paraffin-embedded sections and staining techniques. Hormone levels, including luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), 17-β-hydroxysteroid dehydrogenase 3 (HSD17b3), and follicle-stimulating hormone (FSH), were detected using ELISA.

[0045] 2. Experimental Results

[0046] (1) Ligusticum lactone I can significantly restore the estrous cycle in POI rats.

[0047] Menstrual disorders are one of the most typical clinical manifestations in patients with porcine ovarian hyperemesis gravidarum (POI), manifesting as infrequent menstruation, scanty menstruation, or even amenorrhea. POI rats lack a menstrual cycle, exhibiting estrous cycle irregularities and prolonged interestrous periods. For example... Figure 3 As shown, the normal control group maintained a regular estrous cycle, while the model group exhibited a continuous interestrous period, no longer showing estrus, proestrus, and metestrus. In the positive intervention control group given Fer-1 and the ligustrazine I group, estrus gradually recovered starting 14 days after intervention, and by approximately 28 days post-intervention, estrus cycles had fully recovered, with clear and regular proestrus, estrus, and metestrus manifestations. This invention directly demonstrates that ligustrazine I can effectively improve POI and restore normal estrous cycles in rats. Figure 2 As shown, microscopic observation of the cellular composition of rat vaginal secretions reveals the following: During proestrus, spindle-shaped epithelial cells are distributed among leukocytes, indicating the approach of estrus. During estrus, numerous leukocytes are observed, with no other cells present. During mesestrus, various cell types are visible, including leukocytes, anucleated epithelial cells, and spindle-shaped epithelial cells. During estrus, numerous large, irregularly shaped, anucleated squamous epithelial cells are observed, and leukocytes disappear.

[0048] (2) Ligusticum lactone I can significantly increase angiogenesis in POI rats and improve the developmental morphology of follicles at all stages in the ovary.

[0049] like Figure 4As shown, in the normal control group, ovarian angiogenesis was increased, and the ovarian tissue morphology was regular, with follicles at different developmental stages visible. In the model group, the ovarian index and uterine index of rats were significantly decreased. HE staining showed a significant reduction in the number of total follicles, primordial follicles, growing follicles, and corpora lutea, while the number of atretic and cystic dilated follicles was significantly increased. The ovarian structure was disordered and the volume was small, with loose and disordered granulosa cells, plasma cell infiltration in atretic follicles, and decreased ovarian reserve. In the natural recovery group, the ovarian tissue structure was irregular, the granulosa cell layer was thin, and there were fewer primordial follicles, growing follicles, and mature follicles, and more atretic follicles. In the Fer-1 group, more growing follicles were visible. After treatment with ligustrazine lactone I, the ovarian tissue structure was improved, with more growing follicles visible and the granulosa cell layer thickened.

[0050] (3) Ligusticum lactone I can significantly regulate serum sex hormone levels in POI rats.

[0051] Currently, commonly used laboratory indicators for assessing ovarian reserve function include luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), 17-β-hydroxysteroid dehydrogenase 3 (HSD17b3), and follicle-stimulating hormone (FSH). Basal E2 is the serum E2 level on days 2-4 of the menstrual cycle. E2 is mainly secreted by the granulosa cells of growing follicles and can be used as an indicator for assessing ovarian reserve function and monitoring follicle growth and development. In animal reproductive and endocrine experiments, to avoid errors caused by changes in serum sex hormone levels throughout the estrous cycle, samples are collected and serum sex hormone levels are measured during the interestrous period when levels are lower. Blood is collected from the heart of rats, and the serum levels of various indicators are detected using ELISA. Figure 5 As shown, the levels of E2, AMH, and HSD17b3 in the model group were significantly lower than those in the normal control group, while the level of LH was significantly higher (P<0.01). Compared with the model group, both the Fer-1 group and the Ligusticum striatum lactone I group increased the levels of E2, AMH, and HSD17b3 in rat serum and decreased the levels of LH and FSH (P<0.05 or P<0.01).

[0052] (4) Ligusticum lactone I can significantly improve endometrial thickness and glandular development in POI rats.

[0053] Compared with the normal control group, the model group had poorer endometrial regeneration, thinner endometrium and myometrium, while the Ligusticum chuanxiong lactone I group and Fer-1 group had more glands in the uterus and thicker myometrium. Figure 6 ).

[0054] (5) Ligusticum lactone I can significantly increase the number of offspring produced in POI rats.

[0055] After 28 days of intervention, the rats were placed in cages with sexually mature, fertile male rats to allow for natural conception. The litter size was then recorded, and the results are shown below. Figure 7Compared with POI, the average litter size of the ligustrazine I group was significantly higher, and the time to conception after mating was comparable to that of the normal group, suggesting that ligustrazine I treatment can improve the litter size of POI rats.

[0056] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Application of ligustrolactone I in the preparation of drugs for improving ovarian dysfunction and / or ovarian insufficiency.

2. The application according to claim 1, characterized in that, The drug improves decreased ovarian function and / or ovarian insufficiency by improving ovarian angiogenesis.

3. The application according to claim 1, characterized in that, The drug improves decreased ovarian function and / or ovarian insufficiency by improving the developmental morphology of follicles at all stages.

4. The application according to claim 1, characterized in that, The drug improves decreased ovarian function and / or ovarian insufficiency by increasing ovarian reserve.

5. The application according to claim 1, characterized in that, The drug improves decreased ovarian function and / or ovarian insufficiency by regulating sex hormone levels.

6. The application according to claim 1, characterized in that, The drug improves decreased ovarian function and / or ovarian insufficiency by improving endometrial thickness and glandular development.

7. A drug for improving decreased ovarian function and / or ovarian insufficiency, characterized in that, The active ingredient includes ligustilide I.

8. The medicament according to claim 7, characterized in that, The drug also includes pharmaceutically acceptable excipients.

9. The medicament according to claim 8, characterized in that, The excipients include fillers, binders, disintegrants, emulsifiers, flavoring agents, preservatives, or colorants.

10. The medicament according to claim 7, characterized in that, The dosage form of the drug is tablets, granules, capsules, pills, or oral liquid.

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