Composition containing stem cells and application of composition in ovarian anti-aging

Abstract

The invention relates to a composition containing stem cells and application of the composition in ovarian aging resistance. According to the invention, the polypeptide TP-TOF with ovarian aging resistance is separated and screened from placenta, and the polypeptide can effectively reduce apoptosis of ovarian granular cells. The polypeptide can effectively promote recovery of ovarian estradiol and follicular poietin levels of rats independently or in combination with stem cells, can remarkably prolong the estrus period, reduce the estrus interval, reduce follicle apoptosis and promote follicle generation, and has a wider application prospect.

Description

Technical Field

[0001] This application relates to the field of biology, and more specifically to compositions comprising stem cells and their use in ovarian anti-aging. Background Technology

[0002] Premature ovarian failure (POF) refers to the natural cessation of menstruation in women before the age of 40. The prevalence of POF in the general population is 1%-3%, while 2%-10% of amenorrhea patients have POF. POF is a common disease in the field of gynecological endocrinology, and its incidence has been increasing in recent years. Patients with POF face a range of health problems, including infertility and menopausal syndrome. The etiology of POF is complex, with idiopathic POF accounting for 81% of hypergonadotropic amenorrhea cases.

[0003] Premature ovarian failure (POF) has multiple causes. Genetic factors include several genes associated with POF. FOXL2 gene mutations can lead to abnormal ovarian development and follicular atresia; PRDM1 gene mutations can affect ovarian function, leading to premature menopause. Androgen receptor genes and bone morphogenetic protein 15 genes are also related to POF. Immune factors also play a role in the pathogenesis of POF. Autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) can cause ovarian inflammation, thus affecting ovarian function. Alterations in the ovarian immune microenvironment can also lead to POF. Environmental factors include long-term exposure to certain chemicals such as plasticizers and heavy metals, which can induce POF by damaging ovarian tissue. Lifestyle habits such as excessive weight loss, chronic stress, and smoking can also negatively impact ovarian function. Lifestyle is also a significant factor influencing POF. Long-term sleep deprivation, lack of exercise, and poor dietary habits can lead to decreased immunity and affect ovarian function. Therefore, maintaining healthy lifestyle habits is crucial for preventing POF.

[0004] Treatment for premature ovarian failure (POI) primarily involves various strategies, including hormone replacement therapy, gene therapy, immunotherapy, and lifestyle modifications. Hormone replacement therapy is a commonly used treatment. Because POI patients have low estrogen levels, exogenous hormone replacement therapy can effectively alleviate symptoms and prevent and improve symptoms and complications caused by POI. Estrogen can be administered orally or by injection, helping to regulate the menstrual cycle, protect the endometrium, and prevent osteoporosis. However, long-term use of hormone replacement therapy also requires caution regarding potential side effects, such as breast cancer, endometrial hyperplasia, and even cancer. In addition to hormone replacement therapy, significant progress has been made in gene therapy research for POI in recent years. Through in-depth exploration of genetic factors, scientists have discovered that some cases of POI are related to gene defects or abnormal expression. For these patients with a potential genetic predisposition, targeted gene therapy can be attempted to restore ovarian function, provided that a risk assessment is conducted. Gene editing technology repairs specific gene mutations in ovarian tissue, aiming to restore ovarian function. Immunotherapy has also shown potential in the treatment of POI. Ovarian damage and inflammation caused by autoimmune factors are considered one of the important mechanisms of premature ovarian failure (POI). Regulating immune system function may be beneficial for POI patients. Currently, some experimental immunotherapies have shown effectiveness in restoring ovarian function in clinical trials. The application of drug therapy in premature ovarian failure is becoming increasingly diversified; doctors will choose the most appropriate treatment method based on the specific circumstances of each patient. Whether it's hormone replacement therapy, gene therapy, or immunotherapy, extensive clinical research is still needed to verify their effectiveness and safety. In addition to individualized treatment, lifestyle adjustments are also indispensable, especially maintaining a healthy lifestyle, a reasonable diet, and moderate exercise, all of which can help alleviate premature ovarian failure symptoms and improve quality of life.

[0005] In recent years, with the development of science and technology and the progress of medicine, new technologies combined with therapy have shown broad prospects in the treatment of premature ovarian failure (POF). In particular, stem cell technology, gene editing technology, and endocrine disruptor regulation technology, which have developed in recent years, have provided new ideas and methods for solving this challenging problem. Stem cell technology has enormous potential in the treatment of POF. Stem cells have the ability to self-renew and differentiate into other cell types. By transplanting stem cells into the ovaries of POF patients, it can help restore ovarian function and promote follicle growth and development. Mesenchymal cell transplantation is also a promising treatment method, as it can provide more follicle growth factors and supporting cells, thereby improving ovarian function. Gene editing technologies such as CRISPR / Cas9 have also played an important role in the treatment of POF. By precisely editing genes, scientists can more accurately locate and repair gene mutations that cause POF, thereby fundamentally treating the disease. Although new technologies are emerging one after another, their complexity and high cost mean that truly effective drugs still require further advancement.

[0006] Studies have found that placental peptides, small-molecule active functional peptides extracted from the placenta, can scavenge free radicals, regulate immunity, and inhibit peroxidation reactions. Placental peptide injections can be used to treat diseases caused by decreased or disordered cellular immune function, postoperative healing or viral infections, and leukopenia caused by various reasons. Basic research has found that placental peptide injections can improve ovarian function in rats with diminished ovarian reserve. Clinical studies have also shown that placental peptide injections can improve sex hormone levels and clinical symptoms in patients with diminished ovarian reserve. However, research on placental peptides is still insufficient, especially on highly active peptides, which requires further investigation. Summary of the Invention

[0007] In one aspect, the present invention isolates a highly active peptide with anti-premature ovarian failure activity from the placenta of a sika deer. The peptide is a TP-TOF peptide, and its amino acid sequence is shown in SEQ ID NO: 1.

[0008] Furthermore, the TP-TOF peptide exhibits good anti-apoptotic effects on ovarian granulosa cells, thereby treating premature ovarian failure.

[0009] Furthermore, the present invention provides a pharmaceutical composition for treating premature ovarian failure, which contains a highly active anti-premature ovarian failure peptide as the sole active ingredient, wherein the peptide is a TP-TOF peptide and its amino acid sequence is shown in SEQ ID NO: 1.

[0010] Furthermore, the composition also contains a pharmaceutically acceptable carrier or excipient.

[0011] Specifically, excipient types include: fillers or carriers of active ingredients (e.g., calcium carboxylate, microcrystalline cellulose, corn starch, silica, or calcium carbonate), thickeners, film-forming agents, and binders (e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose, gum arabic, sodium alginate, xanthan gum, and gelatin), buffers and pH control agents (e.g., magnesium oxide, magnesium hydroxide, potassium carbonate, sodium carbonate, potassium bicarbonate, sodium bicarbonate, or mixtures thereof), anti-adhesion agents (e.g., talc), flow aids (e.g., colloidal silica), and natural or artificial sweeteners. Examples of nicotine-containing products include saccharin, acesulfame potassium, aspartame, sucralose, isomaltitol, lactose, mannitol, sorbitol, xylitol, and sucrose; humectants (e.g., glycerin); preservatives and antioxidants (e.g., sodium benzoate and ascorbyl palmitate); surfactants (e.g., polysorbate 80); natural or artificial flavorings (e.g., peppermint, cinnamon, cherry, or other fruit flavorings); dyes or pigments (e.g., titanium dioxide or D&C Yellow No. 10); and lubricants or processing aids (e.g., calcium stearate or magnesium stearate). Certain types of nicotine-containing products may also have a coating composed of ingredients capable of providing an acceptable coating (e.g., the coating may consist of ingredients such as carnauba wax and pharmaceutically acceptable forms of shellac, glazing compositions, and glazing agents).

[0012] In some embodiments, the pharmaceutical composition further includes an inactive ingredient. Specifically, the inactive ingredient is a base or buffer, or a combination thereof, buffered within an alkaline pH range. In other embodiments, the inactive ingredient is an acid or buffer, or a combination thereof, buffered within an acidic pH range.

[0013] The alkaline substance of the inactive component can vary. Exemplary strong bases are sodium hydroxide, potassium hydroxide, and mixtures thereof. Exemplary weak bases are sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, and mixtures thereof. The inactive component, which is combined with a carrier in a close contact manner (e.g., adsorbed onto a porous particulate material (such as microcrystalline cellulose)), can be used as a single component (e.g., as sodium hydroxide or as sodium bicarbonate) or as a combination of at least two components (e.g., as a mixture of sodium carbonate and sodium bicarbonate). Additionally, the inactive component, which is combined with a carrier in a close contact manner (e.g., adsorbed onto a material such as microcrystalline cellulose), can be used as a single buffer component (e.g., as sodium dihydrogen phosphate) or as a combination of at least two components (e.g., as a mixture of sodium carbonate and sodium dihydrogen phosphate).

[0014] Furthermore, non-limiting examples of suitable buffers include aluminum hydroxide, magnesium hydroxide, aluminum hydroxide / magnesium hydroxide co-precipitate, aluminum hydroxide / sodium bicarbonate co-precipitate, calcium acetate, calcium bicarbonate, calcium borate, calcium carbonate, calcium bicarbonate, calcium citrate, calcium gluconate, calcium glycerophosphate, calcium hydroxide, calcium lactate, calcium phthalate, calcium phosphate (including calcium dihydrogen phosphate, tricalcium phosphate, etc.), calcium succinate, calcium tartrate, calcium formate, calcium propionate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, dipotassium phosphate, disodium hydrogen phosphate, disodium succinate, dry aluminum hydroxide colloid, L-arginine, magnesium acetate, magnesium aluminate, magnesium borate, magnesium bicarbonate, magnesium carbonate, magnesium citrate, and gluconate. Magnesium gluconate, magnesium hydroxide, magnesium lactate, magnesium aluminosilicate, magnesium oxide, magnesium phthalate, magnesium phosphate, magnesium silicate, magnesium succinate, magnesium tartrate, potassium acetate, potassium carbonate, potassium bicarbonate, potassium borate, potassium citrate, potassium metaphosphate, potassium phthalate, potassium phosphate, potassium polyphosphate, potassium pyrophosphate, potassium succinate, potassium tartrate, sodium acetate, sodium bicarbonate, sodium borate, sodium carbonate, sodium citrate, sodium gluconate, sodium hydrogen phosphate, sodium hydroxide, sodium lactate, sodium phthalate, sodium phosphate, sodium polyphosphate, sodium pyrophosphate, sodium sesquicarbonate, sodium succinate, sodium tartrate, sodium tripolyphosphate, synthetic hydrotalcite, tetrapotassium pyrophosphate, tetrasodium pyrophosphate, tripotassium phosphate, trisodium phosphate, and tromethamine. Furthermore, due to the ability of proteins or protein hydrolysates to react with gastric acid, they can also act as buffers in embodiments of the invention. Additionally, combinations or mixtures of the above buffers can be used in the pharmaceutical formulations described herein.

[0015] The acidic substance of the inactive component can vary. Exemplary acidic materials include citric acid, malic acid, oxalic acid, levulinic acid, and mixtures thereof. Exemplary buffers include sodium citrate, sodium acetate, monopotassium phosphate, etc. The inactive component, which is combined with a carrier in a close contact manner (e.g., adsorbed onto a porous particulate material (such as microcrystalline cellulose)), can be used as a single component (e.g., as citric acid or as malic acid) or as a combination of at least two components (e.g., as a mixture of malic acid and citric acid). Additionally, the inactive component, which is combined with a carrier in a close contact manner (e.g., adsorbed onto a material such as microcrystalline cellulose), can be used as a single buffer component (e.g., as sodium citrate) or as a combination of at least two components (e.g., as a mixture of sodium citrate and citric acid).

[0016] The amount of inactive components (e.g., basic materials and / or buffers, or acidic materials and / or buffers) adsorbed on the carrier or substrate material can vary. Typically, the substrate material (i.e., a porous particulate carrier material) acting as a carrier for the basic material and / or buffer (or acidic material and / or buffer) is the dominant component (by weight) of the resulting mixture. Typically, based on the combined weight of the substrate material and the basic material and / or buffer (or the combined weight of the substrate material and the acidic material and / or buffer), the substrate material constitutes at least about 70%, typically at least about 80%, frequently at least about 90%, and often at least about 95% of the mixture weight; while based on the combined weight of the substrate material and the basic material and / or buffer (or the combined weight of the substrate material and the acidic material and / or buffer), the amount of the basic material and / or buffer typically constitutes at most about 30%, typically at least about 20%, frequently at most about 10%, and often at most about 5% of the mixture weight.

[0017] The dosage of the active ingredient polypeptide is an amount that is effective in treating, or preventing, certain symptoms of a condition, disease, or disorder suffered by the subject or patient. An "effective amount," "therapeutic amount," or "effective dose" means an amount sufficient to produce the desired pharmacological or therapeutic effect, thereby resulting in effective prevention or treatment of the condition, disease, or disorder. Therefore, an effective amount of the active ingredient is an amount sufficient to enter the relevant area of ​​the body (e.g., including through the subject's ovaries).

[0018] In the pharmaceutical compositions provided herein, any suitable excipients and / or carriers are optionally combined with (e.g., a therapeutically effective amount) a pharmaceutical agent (e.g., a modified protein or protein variant described herein). In some embodiments, suitable pharmaceutically acceptable carriers include, but are not limited to, water, saline solutions (e.g., NaCl), saline, buffered saline, alcohols, glycerol, ethanol, gum arabic, vegetable oils, benzyl alcohol, polyethylene glycol, gelatin, carbohydrates (e.g., lactose, amylose, or starch), sugars (e.g., mannitol, sucrose, or others), dextrose, magnesium stearate, talc, silica, viscous paraffin, aromatic oils, fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, bovine serum albumin, and combinations thereof. In some embodiments, the pharmaceutical formulation comprises one or more adjuvants (e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorants, flavoring agents, and / or aromatic substances, etc.) that do not react adversely with the active compound and do not interfere with its activity. In some embodiments, a water-soluble carrier suitable for intravenous administration is used. In some embodiments, the pharmaceutical composition or drug comprises a certain amount (typically a small amount) of a wetting agent or emulsifier, or a pH buffer. In some embodiments, the pharmaceutical composition is a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained-release formulation, or powder. In some embodiments, the pharmaceutical composition may be formulated as a suppository having a conventional binder and carrier, such as triglycerides. In some embodiments, oral formulations may include standard carriers such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, polyvinylpyrrolidone, sodium saccharin, cellulose, magnesium carbonate, etc. For example, in some embodiments, compositions typically intended for intravenous administration are solutions in sterile isotonic buffer solutions.

[0019] Beneficial effects This invention isolates and screens TP-TOF, a polypeptide with anti-ovarian aging properties, from the placenta. This polypeptide can effectively reduce apoptosis of ovarian granulosa cells. This polypeptide, alone or in combination with stem cells, can effectively promote the recovery of estradiol and follicle-stimulating hormone levels in rat ovaries, significantly increasing estrus duration, reducing interestrus, decreasing follicular apoptosis, and promoting follicle generation, showing broad application prospects. Attached Figure Description

[0020] Figure 1 The effect of combined peptide and / or stem cell therapy on the estrus or interestrus phase in rats. Detailed Implementation

[0021] Those skilled in the art can refer to the content of this document and appropriately improve the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to implement and apply the technology of this invention. Unless otherwise specified, the methods, equipment, and materials in the following embodiments are all conventional methods, equipment, and materials in the art and are commercially available.

[0022] Example 1: Screening and identification of highly active anti-premature ovarian failure peptides from ovarian polypeptides Fresh sika deer placenta was washed, cut, and 100g of the raw material was added to phosphate buffer (1:2, m / V). The tissue was homogenized and centrifuged for 20 min (12000 r / m). The supernatant was ultrafiltered through an ultrafiltration membrane (retaining peptides with a relative molecular mass less than 3kJ) and separated using DEAE Sepharose CL-6B (2.6cm × 35cm) anion exchange chromatography. The separated components were screened for their anti-apoptotic activity against ovarian granulosa cells. The component with the strongest anti-apoptotic activity, component 4, was further separated using a Sephadex G225 (110cm × 75cm) gel chromatography column. The flow rate was 10 mL / h, and the mobile phase was phosphate buffer. A total of 6 components were separated, among which component 3 showed the strongest anti-apoptotic activity. Component 3 was then separated using a Sephasil C18 (4.6cm × 250mm) reversed-phase high-performance liquid chromatography column. The flow rate was 1 mL / min, and the mobile phase was a 5% acetonitrile solution containing 0.05% (v / v) trifluoroacetic acid. The gradient elution buffer was a 5%–40% acetonitrile solution containing 0.05% trifluoroacetic acid. A peptide with the strongest anti-apoptotic effect on ovarian granulosa cell apoptosis was isolated. Mass spectrometry analysis identified the peptide, and its amino acid sequence is shown in SEQ ID NO: 1. It was named TP-TOF peptide. The peptide was then commissioned to Suzhou Modifu Biotechnology for peptide synthesis. The peptide was 99% pure and its concentration was adjusted to 3 mg / mL for later use.

[0023] Example 2: Effect of TP-TOF peptide on the activity of ovarian granulosa cells SPF mice were subcutaneously injected with PMSG (10 U / mouse) and housed at 23°C with free access to water and food. After 48 hours, the mice were euthanized by cervical dislocation. Both ovaries were rapidly removed under aseptic conditions, washed three times with PBS, and the periovarian capsule and adipose tissue were removed under a stereomicroscope. The ovaries were then placed in DMEM / F-12 culture medium and incubated at 37°C for 15 min. Pre-ovulatory follicles were then punctured with a stainless steel needle under a dissecting microscope to release granulosa cells and oocytes. The granulosa cells were separated from the oocytes by digestion with 1 g / L hyaluronidase. The cells were passed through a 200-mesh screen, centrifuged at 1000 rpm for 5 min, and the supernatant was discarded. The cell pellet was then suspended in DMEM / F-12 culture medium and incubated at 37°C with 50 ml / L CO2 for 15 min. The cells were then washed three times with DMEM / F-12 culture medium and centrifuged at 1000 rpm for 5 min. The viable cell ratio was 97.4% as determined by the trypan blue rejection test. Cells were diluted with a certain amount of serum-free DMEM / F-12 medium (containing 100 U / L penicillin, 100 μg / mL streptomycin, and 0.5 g / L LBSA) and counted using the hemocytometer. Cell density was adjusted, and cells were seeded into two 250 mL sterile culture flasks and pre-cultured in a 37°C, 50 mL / L CO2 incubator.

[0024] Cells were divided into a blank control group (containing 0.5 g / L BSA in DMEM / F-12 medium) and experimental groups (containing 10 μg / mL, 50 μg / mL, 100 μg / mL, and 200 μg / mL TP-TOF peptide, respectively), with five replicates in each group. Granulosa cell apoptosis was detected using the TUNEL assay. The apoptosis index of ovarian granulosa cells was calculated as the proportion of positive cells to the total number of cells. The results are shown in Table 1.

[0025] Table 1 Results of ovarian granulosa cell apoptosis index in each group As shown in Table 1, TP-TOF peptide can effectively reduce ovarian granulosa cell apoptosis with increasing dosage, and has a good inhibitory effect on ovarian granulosa cell senescence, thus exhibiting a good therapeutic effect.

[0026] Example 3: Therapeutic Experiment of TP-TOF Peptide Combined with Stem Cells on Premature Ovarian Failure in Rats Rat placental mesenchymal stem cells (Warner Biotech, catalog number WN-87409) were passaged and cultured at a concentration of 1×10⁻⁶. 6 Cell preparation of placental mesenchymal stem cells per ml.

[0027] Sixty healthy female SPF-grade SD rats (10 weeks old, weighing (220±15) g) were housed in a healthy environment at a temperature of 21–25℃ and humidity of 55%–70%, with a 12-hour day-night cycle. The rats had free access to food and water during this period. After one week of acclimatization, they were randomly divided into six groups: a blank control group, a cyclophosphamide model group, a positive control group (estradiol valerate, 0.1 mg / kg), a TP-TOF peptide treatment group (0.1 mg / kg), and a stem cell treatment group (injected with placental mesenchymal stem cell preparation three times a week, 2 × 10⁶ per mouse). 5 (One mouse was injected via tail vein with placental mesenchymal stem cells), TP-TOF peptide combined with placental mesenchymal stem cell treatment group (combined treatment group, 0.1 mg peptide / kg + (injected 3 times a week with placental mesenchymal stem cell preparation, 1×10⁻⁶ per mouse)). 5 (Placement mesenchymal stem cells were injected via tail vein). Fifty rats, excluding the control group, were intraperitoneally injected with cyclophosphamide (60 mg / kg) for 1 day followed by cyclophosphamide (10 mg / kg) for 14 days to establish an induced premature ovarian failure model. TP-TOF peptide was administered via gavage. The positive control group was administered a combination of cyclophosphamide and estrogen solution, while the control group and the cyclophosphamide model group were administered an equal volume of physiological saline via gavage. Administration was once daily for 4 weeks.

[0028] Blood was collected from the tail vein of all the animals four weeks after drug administration, and the levels of estradiol and follicle-stimulating hormone in rat serum were measured using radioimmunoassay. The results are shown in Table 2.

[0029] Table 2. Serum estradiol and follicle-stimulating hormone levels in rats As shown in Table 2, the follicle-stimulating hormone (FSH) levels in the model group mice were significantly higher than those in the blank control group, while estradiol levels were significantly lower. This indicates that CTX administration damages ovarian granulosa cells, reducing estradiol secretion and subsequently increasing FSH levels. After treatment with TP-TOF peptides, the levels of both FSH and estradiol returned to near-normal levels. Stem cell therapy alone can also effectively reduce FSH production and increase estradiol concentration, while the combined treatment of peptides and stem cells showed a better effect, essentially restoring hormone levels to near-normal levels compared to the blank control group.

[0030] Vaginal smear assays were performed twice daily, at 8:00 AM and 8:00 PM. Mice were examined for their estrous cycles under a light microscope. The estrous cycle was analyzed according to standard criteria based on the type of exfoliated cells, with the percentage of estrus or estrus interphase as the composition ratio. Results are as follows: Figure 1 As shown.

[0031] from Figure 1It can be seen that the estrus period in the model group was (9.3±0.7)%, and the interestrus period was (42.5±1.8)%, while the estrus period in the blank control group was (20.7±0.9)%, and the interestrus period was (24.0±1.2)%. This shows that the estrus period in the model group was significantly shorter and the interestrus period was significantly longer compared to the blank control group (P<0.05). In contrast, the positive control group and TP-TOF peptide treatment significantly increased the estrus period and shortened the interestrus period. In particular, the estrus period in the TP-TOF peptide combined with stem cell group was (22.4±1.0)%, and the interestrus period was (22.5±0.4)%, significantly increasing the estrus period and decreasing the interestrus period, showing a significant improvement compared to the positive control group. This fully demonstrates that stem cell combined with TP-TOF peptide can significantly enhance the effect of TP-TOF peptide.

[0032] After the experiments were completed, the rats in each group were sacrificed, and the uterus and bilateral ovaries were removed. The bilateral ovaries were fixed with 4% paraformaldehyde, routinely embedded in paraffin, and sectioned into 5 μm sections. The sections were stained with hematoxylin and eosin (HE) and observed under a light microscope to observe the morphological changes of the bilateral ovarian tissue. The number of follicles with normal follicular structures was counted. The number of primordial follicles, mature follicles, and total follicles in each group of rats were measured. The results are shown in Table 3.

[0033] Table 3 Primordial follicles, mature follicles, and total number of follicles in rats. As shown in Table 3, the number of primordial follicles, mature follicles, and total follicles in the model group were significantly lower than those in the blank control group. In contrast, the corresponding number of primordial follicles, mature follicles, and total follicles in each treatment group were significantly increased. In particular, after TP-TOF peptide treatment, the number of mature follicles and total follicles significantly increased. Furthermore, TP-TOF peptide combined with stem cell therapy also significantly increased the number of mature follicles and total follicles compared to TP-TOF peptide alone, indicating a good synergistic therapeutic effect. These results demonstrate that the TP-TOF peptide of this invention has the effect of reducing follicle apoptosis and promoting follicle generation, and its application prospects are even broader after combination with stem cell therapy.

[0034] Although embodiments of the present invention have been described above in conjunction with the accompanying drawings, the present invention is not limited to the specific embodiments and application fields described above. The specific embodiments described above are merely illustrative and instructive, and not restrictive. Those skilled in the art can make many other forms based on the guidance of this specification and without departing from the scope of protection of the claims of the present invention, and all of these are within the scope of protection of the present invention.

Claims

1. A high activity anti-ovarian premature aging active peptide TP-TOF, characterized in that The amino acid sequence is shown in SEQ ID NO:

1.

2. Use of the high-activity anti-ovarian premature senility active peptide TP-TOF in the preparation of a drug for treating ovarian premature senility according to claim 1.

3. Use of the high-activity anti-ovarian premature senility active peptide TP-TOF and placental mesenchymal stem cells in the preparation of a kit for treating ovarian premature senility according to claim 1.

4. The use according to claim 2, characterized in that The drug contains a pharmaceutically acceptable carrier or excipient.

5. The use according to claim 2, characterized in that The placental mesenchymal stem cells are commercially purchased.

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