Application of hippocampus multi-penis pill in preparation of medicine for improving or treating female infertility

Abstract

The invention belongs to the technical field of new application of hippocampus multi-penis pills, and provides application of the hippocampus multi-penis pills in preparation of a medicine for improving or treating female infertility. Animal test results show that the hippocampus multi-penis pill has a good treatment effect on premature ovarian failure and endometriosis. Clinical test results show that when the hippocampus multi-penis pill is applied to female infertility patients, the treatment effect is remarkable, and the development of ovarian follicles can be effectively promoted, the endometrial receptivity can be improved, the immune function of the organism can be regulated, and the inflammation can be inhibited. Animal test and clinical test results show that the hippocampus multi-penis pill has a certain application prospect in preparation of medicines for improving or treating female infertility.

Description

Technical Field

[0001] The present invention belongs to the technical field of new applications of Haima Duobian pills, and in particular relates to the application of Haima Duobian pills in the preparation of medicines for improving or treating female infertility. Background Art

[0002] Female infertility refers to a reproductive health problem in which the woman of childbearing age is unable to conceive naturally within one year without taking any contraceptive measures and having normal sexual life. It has become a major public health challenge affecting family happiness and population development worldwide.

[0003] With the influence of factors such as the accelerated pace of life in modern society, increased work pressure, intensified environmental pollution and delayed childbearing age, the incidence of female infertility has been increasing year by year. Some unexplained or intractable infertility has brought huge physical and mental burdens to patients.

[0004] From an etiological perspective, the pathogenesis of female infertility is complex and diverse, involving abnormalities in multiple physiological processes. Common causes include ovarian dysfunction such as ovulatory disorders and corpus luteum insufficiency; fallopian tube dysfunction resulting from blockage, adhesions, and hydrops leading to impaired patency or peristalsis; uterine factors such as endometriosis, endometrial polyps, and intrauterine adhesions; and immune rejection reactions such as antisperm antibodies, antiphospholipid antibody syndrome, and blocking antibody deficiency. Furthermore, endocrine disorders such as polycystic ovary syndrome, hyperprolactinemia, and thyroid dysfunction, genetic factors, and unhealthy lifestyle factors can interfere with female reproductive function through various pathways, leading to difficulty conceiving.

[0005] At present, clinical treatments for female infertility mainly include Western medicine treatment, surgical treatment and assisted reproductive technology.

[0006] Western medicine treatments often focus on symptomatic interventions, such as using ovulation-inducing drugs like clomiphene and letrozole to improve ovulatory disorders, using progesterone supplementation to treat luteal insufficiency, or using immunosuppressants like glucocorticoids and anticoagulants like low-molecular-weight heparin to regulate immune disorders. However, Western medicine treatments have certain limitations. For example, ovulation-inducing drugs may trigger ovarian hyperstimulation syndrome, and long-term use of immunosuppressants can easily lead to a decrease in the body's immunity. In addition, some patients do not respond well to medication, resulting in suboptimal treatment results.

[0007] Surgical treatment is mainly used to correct organic lesions, such as fallopian tube dredging and endometrial polypectomy, but surgical trauma may increase the risk of pelvic adhesions, and has limited efficacy for infertility caused by functional or immune factors.

[0008] Although assisted reproductive technologies such as in vitro fertilization provide some refractory patients with the possibility of fertility, they have problems such as high costs, large fluctuations in success rates, painful treatment processes, and increased risks of multiple pregnancies, making them difficult to be widely popularized in clinical practice.

[0009] Traditional Chinese Medicine (TCM) has a long history and unique advantages in treating female infertility. It regulates the body's yin and yang balance, improves organ function, and dredges the Qi and blood vessels, achieving the goal of regulating menstruation. It boasts minimal side effects, high safety, and good patient tolerance.

[0010] Haima Duobian Pills are a traditional Chinese medicine pill made from 34 Chinese medicinal materials including seahorse, gecko, deer antler, dog whip, etc. It has the effects of nourishing the kidney and strengthening yang, replenishing essence and nourishing marrow. It is suitable for symptoms such as deficiency of qi and blood, soreness of waist and legs, impotence and premature ejaculation. Summary of the Invention

[0011] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide an application of Haima Duobian Pills in the preparation of a drug for improving or treating female infertility.

[0012] In order to achieve the above object, the solution adopted by the present invention is:

[0013] Application of Haima Duobian Pills in the preparation of medicines for improving or treating female infertility.

[0014] Furthermore, in a preferred embodiment of the present invention, infertility includes infertility caused by ovarian dysfunction, infertility caused by uterine abnormality, infertility caused by fallopian tube dysfunction, infertility caused by immune disorder, and infertility caused by endocrine disorder.

[0015] Furthermore, in a preferred embodiment of the present invention, ovarian dysfunction includes luteal insufficiency, premature ovarian failure and hypothalamic anovulation.

[0016] Furthermore, in a preferred embodiment of the present invention, fallopian tube dysfunction includes weakened or disordered fallopian tube motility caused by chronic salpingitis, and ciliary loss or weak swinging caused by damage to the fallopian tube mucosa.

[0017] Furthermore, in a preferred embodiment of the present invention, the uterine abnormality includes endometriosis.

[0018] Furthermore, in a preferred embodiment of the present invention, the immune disorder includes anti-sperm immune disorder, anti-endometrial immune disorder, anti-ovarian immune disorder and anti-zona pellucida immune disorder.

[0019] Furthermore, in a preferred embodiment of the present invention, endocrine disorders include hyperandrogenism, abnormal estrogen secretion and abnormal thyroid function.

[0020] Application of Haima Duobian Pills in the preparation of auxiliary medicines for improving or treating female infertility.

[0021] The invention relates to the use of Haima Duobian pills in combination with other drugs in the preparation of drugs for improving or treating female infertility.

[0022] The beneficial effects of the application of the Haima Duobian pill provided by the present invention in the preparation of a drug for improving or treating female infertility are:

[0023] In this application, the results of animal experiments on premature ovarian failure show that Haima Duobian Pills have a significant improvement effect on the estrous cycle disorders caused by chemotherapy-induced premature ovarian failure caused by cisplatin, and have a certain protective and improving effect on the uterus of model rats. It can have a significant regulatory effect on the hormone level disorders caused by premature ovarian failure, and can also improve ovarian reserve function.

[0024] In this application, the results of endometriosis animal experiments showed that after administration of Haima Duobian Pills, the volume of ectopic endometrium in rats was significantly reduced, and the levels of serum PGE2 and PGF2α were significantly decreased, indicating that Haima Duobian Pills has a good therapeutic effect on endometriosis.

[0025] In this application, the clinical trial results show that the treatment effect of Haima Duobian Pills on female infertility patients is significant, and it can effectively promote the development of ovarian follicles, improve endometrial receptivity, regulate the body's immune function, and inhibit inflammation and other reactions.

[0026] The results of animal experiments and clinical trials show that Haima Duobian Pills have certain application prospects in the preparation of treatments for improving or treating female infertility. DETAILED DESCRIPTION

[0027] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0028] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0029] Example 1

[0030] This experimental example verifies the effect of Haima Duobian Pills on premature ovarian failure based on a rat model of chemotherapy-induced premature ovarian failure caused by cisplatin.

[0031] 1. Experimental Animals: 40 healthy, non-pregnant female SD rats weighing 200-220 g were selected and housed in a conventional manner for 3 days. During the acclimatization period, the rats were allowed free access to water and food at room temperature of 22°C-26°C and under natural light.

[0032] 2. Experimental methods:

[0033] 2.1 Grouping and Dosing:

[0034] Grouping: The mice were randomly divided into 4 groups according to body weight, with 10 mice in each group, namely: blank group, model group, estradiol valerate tablets group (dose 0.1 mg / kg), and Haima Duobian pills group (dose 0.1 mg / kg).

[0035] Dosage:

[0036] Blank group: No model was established, purified water was given orally, 10 mg / kg -1 ;

[0037] Model group: Purified water was administered orally, 10 mg / kg -1 ;

[0038] Estradiol valerate tablet group: Estradiol valerate was administered orally, with a dosage of 10 mg / kg -1 ;

[0039] Haima Duobian pill group: Haima Duobian pill was administered orally at a volume of 10 mg / kg. -1 .

[0040] 2.2 Model Establishment: After 14 days of preventive administration, rats in all groups except the blank group were given an animal model of premature ovarian failure. Cisplatin (dissolved in saline to prepare a 1 mg / mL solution) was injected intraperitoneally at 4 mg / kg on days 14 and 21, respectively. Rats in each treatment group were given the drug by gavage continuously until the end of the experiment. At the end of the experiment, the rats were anesthetized with an intraperitoneal injection of 10% chloral hydrate 3 mL / kg. Blood was collected through the abdominal aorta and centrifuged at 3500 r / min for 10 minutes. The serum was frozen and stored in a -80°C refrigerator. The model was successfully established.

[0041] 2.3 Test indicators and results:

[0042] 2.3.1 Effects on the estrous cycle of rats

[0043] Estrous cycle detection of rats: Starting from the second injection of cisplatin, vaginal exfoliated cell smears were performed at 9 am every day for 10 consecutive days. The changes in the estrous cycle of the rats were observed and recorded under an optical microscope. The experimental results are shown in Table 1 below.

[0044] Table 1

[0045]

[0046]

[0047] Note: Compared with the blank group, #P<0.05, compared with the model group, *P<0.05.

[0048] The results in Table 1 show that the model group rats exhibited significant estrous cycle disruption, manifested as absent estrus or prolonged diestrus, with significant differences compared to the blank group (p < 0.05). The number of rats with estrous cycle disruption in the Haima Duobian Pill group was significantly reduced compared to the model group (p < 0.05). These results suggest that Haima Duobian Pill has a significant ameliorative effect on estrous cycle disruption caused by chemotherapy-induced premature ovarian failure (CIPF) induced by cisplatin.

[0049] 2.3.2 Effects on rat organ coefficients

[0050] Organ coefficient test: The uterus and bilateral ovaries of rats were removed, the surrounding adipose tissue was removed, and the weights were accurately weighed using an electronic balance. The organ coefficient was calculated [organ coefficient (%) = organ wet weight (g) / body weight (g) × 100%]. The experimental results are shown in Table 2 below.

[0051] Table 2

[0052] Group Ovarian coefficient (%) Uterine coefficient (%) Blank group 0.044±0.007 0.332±0.112 Model Group <![CDATA[0.032±0.006 # ]]> <![CDATA[0.205±0.104 # ]]> Estradiol valerate tablets <![CDATA[0.039±0.008 * ]]> <![CDATA[0.315±0.115 * ]]> Haima Duobian pill group <![CDATA[0.041±0.007 * ]]> <![CDATA[0.319±0.108 * ]]>

[0053] Note: Compared with the blank group, # P<0.05, compared with the model group, *P<0.05.

[0054] The results in Table 2 show that the ovarian and uterine coefficients of the model rats were significantly reduced, with significant differences compared to the blank group (p < 0.05), suggesting that the model rats had ovarian and uterine damage. The uterine and ovarian coefficients of the rats in the Haima Duobian Pill group were significantly increased compared to the model group, with significant differences (p < 0.05). These results suggest that Haima Duobian Pill has a protective effect on the uterus of the model rats.

[0055] 2.3.3 Effects on serum sex hormone levels in rats

[0056] Serum hormone level detection: ELISA method was used in strict accordance with the kit instructions to determine the levels of estrogen, gonadotropin and other related hormones in serum. The experimental results are shown in Table 3.

[0057] Table 3

[0058] Group E2 (pg / mL) AMH (ng / mL) FSH (ng / mL) LH (mLU / mL) Blank group 125.08±22.82 10.48±3.25 0.25±0.06 10.31±3.65 Model Group <![CDATA[50.86±18.33 # ]]> <![CDATA[5.04±2.32 # ]]> <![CDATA[0.62±0.12 # ]]> <![CDATA[25.31±8.78 # ]]> Estradiol valerate tablets <![CDATA[88.32±15.35 * ]]> <![CDATA[10.20±3.11 * ]]> <![CDATA[0.15±0.15 * ]]> <![CDATA[9.05±4.88 * ]]> Haima Duobian pill group <![CDATA[85.64±19.22 * ]]> <![CDATA[9.84±2.24 * ]]> <![CDATA[0.21±0.08 * ]]> <![CDATA[10.55±4.26 * ]]>

[0059] Note: Compared with the blank group, # P<0.05, compared with the model group, *P<0.05.

[0060] The results in Table 3 showed that the serum E2 and AMH levels of the rats in the model group were significantly decreased, and the FSH and LH levels were significantly increased, which were significantly different from those in the blank group (all p < 0.05), indicating that the serum hormone levels of the rats in the chemotherapy-induced premature ovarian failure model induced by cisplatin were disordered and the ovarian reserve function was decreased.

[0061] Compared with the model group, the serum E2 and AMH levels of rats in the estradiol valerate tablet group were significantly increased, while the FSH and LH levels were significantly decreased, with significant differences (all p < 0.05). The serum E2 and AMH levels of rats in the Haima Duobian pill group were significantly increased, while the FSH and LH levels were significantly decreased, also with significant differences compared with the model group (p < 0.05). These results suggest that Haima Duobian pill has a significant regulatory effect on hormonal imbalances caused by chemotherapy-induced premature ovarian failure induced by cisplatin and can also improve ovarian reserve function.

[0062] 2.3.4 Effects on rat ovarian histopathology

[0063] Ovarian histopathological examination: The ovaries were fixed in paraformaldehyde for 48 h, dehydrated, transparentized, wax-impregnated, embedded, sectioned, dewaxed, and stained with HE. The morphological changes of the ovarian tissue were observed under an optical microscope, and the total number of follicles and the number of atretic follicles were counted. The experimental results are shown in Table 4 below.

[0064] Table 4

[0065] Group Total number of follicles Number of atretic follicles Blank group 70.02±10.02 0.00±0.00 Model Group <![CDATA[30.22±4.88 # ]]> <![CDATA[6.32±1.24 # <!-- 4 -->]]> Estradiol valerate tablets 53.48±6.25* 2.24±1.08* Haima Duobian pill group 51.66±7.04* 2.86±0.86*

[0066] Note: Compared with the blank group, # P<0.05, compared with the model group, *P<0.05.

[0067] The experimental results showed pathological changes in the ovarian tissue of the model rats, with a significant decrease in total follicles and an increase in atretic follicles. The Haima Duobian Pill group showed a significant increase in total follicles and a decrease in atretic follicles compared to the model group (p < 0.05). These results suggest that Haima Duobian Pill has a beneficial effect on cisplatin-induced chemotherapy-induced premature ovarian failure.

[0068] Example 2

[0069] This experimental example was based on endometriosis model rats to verify the effect of Haima Duobian Pills on endometriosis.

[0070] 1. Experimental Animals: 40 SPF-grade, healthy, non-pregnant female Sprague-Dawley rats weighing 200-220 g were selected and housed in a conventionally maintained environment for 1 week. During the acclimatization period, they were allowed free access to water and food. The room temperature was 22°C-26°C and the rats were kept under natural light.

[0071] 2. Experimental methods:

[0072] 2.1 Grouping and Modeling:

[0073] Grouping: The mice were randomly divided into 3 groups according to body weight, with 10 mice in each group, namely: blank group, model group, and Haima Duobian pill group (dose 0.1 mg / kg).

[0074] Model establishment: Rats in the model and treatment groups were anesthetized with an intraperitoneal injection of 2.5% sodium pentobarbital solution. Following fixation, skin preparation, and disinfection, the abdominal cavity was opened midway in the lower abdomen, the uterus was excised, and tissue from the right uterine horn was removed. The endometrium was then stripped. Two 5 mm × 5 mm fragments were obtained using a biopsy punch and transplanted into the mesenteric artery, as far from the incision as possible. The abdominal cavity was irrigated with gentamicin, the abdomen was closed layer by layer, and the incision was sutured. Within 3 days after surgery, 0.1 mL of gentamicin was injected intraperitoneally to prevent infection. Four weeks after modeling, laparotomy examination revealed oval or round cystic nodules ≥ 2 mm in height, with abundant surface vascularization and close adhesion to the surrounding tissue, indicating successful modeling. A total of 20 rats were successfully established. Ten rats were selected as the control group, and only the uterine tissue was excised and sutured.

[0075] 2.2 Administration:

[0076] Drug administration: Start administration 4 weeks after model establishment:

[0077] Blank group: Oral administration of 0.9% sodium chloride solution, dosage volume 10 mg / kg -1 , once a day for 28 consecutive days;

[0078] Model group: Oral administration of 0.9% sodium chloride solution, dosage volume 10 mg / kg -1 , once a day for 28 consecutive days;

[0079] Haima Duobian pill group: Haima Duobian pill was administered orally at a volume of 10 mg / kg. -1 , once a day for 28 consecutive days.

[0080] 2.3 Test indicators and results:

[0081] 2.3.1 Detection of ectopic endometrial lesion volume in rats

[0082] 24 hours after the last administration, the rats were killed, the abdominal cavity was opened, and the volume of the ectopic endometrial lesions in each group of rats was measured and calculated using a vernier caliper. The results of the ectopic endometrial volume of rats after modeling are shown in Table 5.

[0083] Table 5

[0084] Group <![CDATA[Endometrial volume (mm 3 )]]> Blank group <![CDATA[1.35±0.28 # ]]> Model Group <![CDATA[92.21±11.32 # ]]> Haima Duobian pill group <![CDATA[56.64±10.04 * ]]>

[0085] Note: Compared with the blank group, #P<0.05, compared with the model group, *P<0.05.

[0086] The results in Table 4 showed that compared with the blank group, the volume of ectopic endometrium in the model group was significantly increased (P < 0.05); compared with the model group, the volume of ectopic endometrium in the Haima Duobian pill group was significantly decreased (P < 0.05).

[0087] 2.3.2 ELISA method for detection of PGE2 and PGF2α levels in rat serum

[0088] After the rats were killed, blood was collected from the abdominal aorta, placed in a 4°C refrigerator for 3 h, and then centrifuged at 3000 rpm for 15 min (centrifugal radius of 12.5 cm). The supernatant was collected and the levels of PGE2 and PGF2α were detected using ELISA kits. The results are shown in Table 6.

[0089] Table 6

[0090] Group PGE2 (pg / ml) PGF2α (pg / ml) Blank group 118.28±15.28 62.31±6.68 Model Group <![CDATA[148.12±11.34 # ]]> <![CDATA[95.04±12.35 # ]]> Haima Duobian pill group <![CDATA[122.34±10.64 * ]]> <![CDATA[70.26±8.34 * ]]>

[0091] Note: Compared with the blank group, # P<0.05, compared with the model group, *P<0.05.

[0092] The results in Table 6 showed that compared with the normal group, the expression of serum PGE2 and PGF2α in the model group was significantly increased (P < 0.05); compared with the model group, the levels of PGE2 and PGF2α in the Haima Duobian pill group were significantly decreased (P < 0.05).

[0093] After administration of Haima Duobian Pills, the volume of the ectopic endometrium of rats was significantly reduced, and the levels of serum PGE2 and PGF2α were significantly decreased. The results suggest that Haima Duobian Pills has a good therapeutic effect on endometriosis.

[0094] Example 3

[0095] 1. General Information

[0096] Fifty female infertility patients were enrolled as outpatients between January 2024 and March 2025. The oldest patient was 42 years old, the youngest was 21, and the average age was 28.48 years. The shortest duration of infertility was one year, the longest was 12 years, and the average duration was 2.8 years. The causes of infertility included hyperprolactinemia in two cases, premature ovarian failure in 12, endometriosis in 10, luteal insufficiency in 10, endometritis in four, immune infertility in four, and salpingitis in six.

[0097] 2. Case selection:

[0098] 2.1 Inclusion Criteria: ① Married women of childbearing age with normal sexual activity, male reproductive function, and unprotected cohabitation for two years without conception. ② Infertility due to ovarian dysfunction. ③ Infertility due to endometriosis. ④ Infertility due to salpingitis. ⑤ Infertility due to immune disorders. ⑥ Infertility due to endocrine disorders.

[0099] 2.2 Exclusion Criteria: ① Infertility due to congenital physical defects or malformations. ② Infertility due to genetic factors. ③ Infertility due to immune factors. ④ Patients with serious primary cardiovascular, liver, kidney, or hematopoietic system diseases or mental illness. ⑤ Allergic reactions to medications.

[0100] 3. Efficacy judgment:

[0101] Effective: The woman becomes pregnant.

[0102] Ineffective: Symptoms after treatment are not improved compared with those before treatment.

[0103] 4. Dosage: Patients should take the medicine on the third day after menstruation, 2 times / day, 10 tablets / time, for 3 consecutive months.

[0104] 5. Experimental results: Among 50 patients, 27 were effective and 23 were ineffective, with an overall effective rate of 54%.

[0105] The clinical trial results show that the Haima Duobian Pills of the present invention have significant therapeutic effects and can effectively improve the success rate of pregnancy in patients. Haima Duobian Pills have certain application prospects in the preparation of drugs for improving or treating female infertility.

[0106] In summary, in this application, the results of animal experiments show that Haima Duobian Pills have a good therapeutic effect on premature ovarian failure and endometriosis. In this application, the results of clinical trials show that the treatment of female infertility patients with Haima Duobian Pills is effective, and can effectively promote ovarian follicle development, improve endometrial receptivity, regulate the body's immune function, and inhibit inflammation. The results of animal experiments and clinical trials show that Haima Duobian Pills have certain application prospects in the preparation of treatments to improve or treat female infertility.

[0107] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. Application of Haima Duobian Pills in the preparation of drugs for improving or treating female infertility.

2. The use according to claim 1, characterized in that: The infertility includes infertility caused by ovarian dysfunction, infertility caused by uterine abnormality, infertility caused by fallopian tube dysfunction, infertility caused by immune disorder, and infertility caused by endocrine disorder.

3. The use according to claim 2, characterized in that: The ovarian dysfunction includes luteal insufficiency, premature ovarian failure and hypothalamic anovulation.

4. The use according to claim 2, characterized in that: The abnormal fallopian tube function includes weakened or disordered fallopian tube peristalsis caused by chronic salpingitis, and ciliary loss or weak swinging caused by damage to the fallopian tube mucosa.

5. The use according to claim 2, characterized in that: The uterine abnormality includes endometriosis.

6. The use according to claim 2, characterized in that: The immune disorders include anti-sperm immune disorders, anti-endometrial immune disorders, anti-ovarian immune disorders and anti-zona pellucida immune disorders.

7. The use according to claim 2, characterized in that: The endocrine disorders include hyperandrogenism, abnormal estrogen secretion and abnormal thyroid function.

8. Use of Haima Duobian Pills in the preparation of auxiliary medicines for improving or treating female infertility.

9. The use of Haima Duobian Pills in combination with other drugs in the preparation of drugs for improving or treating female infertility.

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