Panax notoginseng saponins Ft1 liposome nanoparticles, and preparation method and application thereof

Abstract

The present application relates to the technical field of medicine, in particular to a kind of panax notoginseng saponin Ft1 liposome nanoparticle and its preparation method and application.The present application selects FSH modified ovary active targeting liposome as the nano-carrier of panax notoginseng saponin Ft1, constructs a kind of new type of ovary protection nano-preparation FSH-Lipo@Ft1.The above preparation can effectively alleviate the weight loss of mouse caused by cisplatin chemotherapy, restores ovarian volume and quality, increases the number of follicle reserve, improves oocyte quality, and plays an obvious protective effect on ovarian tissue structure and overall fertility.The present application can realize the double goals of targeted drug delivery and functional recovery in cisplatin-induced POF, provides a better nano-treatment strategy for ovary protection, and has wide development and application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a liposome nanoparticle containing Panax notoginseng saponin Ft1, its preparation method, and its application. Background Technology

[0002] Ovarian aging is a core biological process of female reproductive system development with age, primarily manifested as diminished ovarian reserve (DOR) and an irreversible decline in the number and quality of oocytes. The ovary is a functional whole composed of multiple cell types, and granulosa cells provide oocytes with nutrients, energy substrates, and growth factors, dynamically maintaining the homeostasis of the ovarian microenvironment, thereby improving follicle development and ovarian function. This offers a novel strategy for treating DOR, and its clinical translational potential is increasingly attracting attention.

[0003] Notoginsenoside Ft1, a rare triterpenoid saponin isolated from Panax notoginseng, has attracted attention in recent years for its pharmacological research in cardiovascular, nervous system, and oncology fields. In addition to its cardiovascular and cerebrovascular protective and anti-inflammatory effects, notoginsenoside Ft1 shows significant advantages in regulating angiogenesis, particularly in improving ovarian microcirculation and ovarian vascular function. However, due to limitations in its water solubility, lipid solubility, and membrane permeability, notoginsenoside Ft1 faces challenges such as low bioavailability and insufficient in vivo exposure after administration. Therefore, there is an urgent need to construct an efficient ovarian-targeted delivery system to overcome the bottlenecks limiting the in vivo application of notoginsenoside Ft1, achieving precise enrichment and sustained effects in ovarian tissue, and providing a novel therapeutic strategy for protecting female fertility.

[0004] Liposomes are a mature nanodelivery platform with good biocompatibility and low immunogenicity, and their safety has been well demonstrated in clinical applications. Using liposomes to load poorly soluble or easily degradable drugs not only improves drug stability but also significantly enhances their bioavailability. Follicle-stimulating hormone (FSH) peptides specifically recognize the highly expressed FSH receptors on the surface of ovarian granulosa cells, possessing advantages such as strong targeting, tissue specificity, good biocompatibility, and low immunogenicity. Modifying the surface of liposomes with FSH peptides can guide the drug delivery system to precisely locate and accumulate in ovarian tissue, reducing non-specific drug distribution in non-target organs, thereby significantly improving the overall effective drug concentration and therapeutic effect in the ovarian region.

[0005] Based on this, the present invention selects liposomes as the nanocarrier of Panax notoginseng saponin Ft1, and innovatively combines Panax notoginseng saponin Ft1 with an active targeted delivery strategy to ovaries to construct a novel ovarian protection nanomedicine system. It aims to solve the above-mentioned existing technical problems and provide a highly efficient and specific new treatment solution for fertility protection in people with diminished ovarian reserve. Summary of the Invention

[0006] The purpose of this invention is to provide a liposome nanoparticle of Panax notoginseng saponin Ft1 and its preparation method.

[0007] Another object of the present invention is to provide the application of the above-mentioned Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of drugs for the prevention or treatment of cisplatin-induced ovarian reserve deficiency.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The liposome nanoparticles of Panax notoginseng saponin Ft1 described in this invention are made from the following components in parts by weight: 35-45 parts soybean lecithin, 2-3 parts cholesterol, 2-3 parts DSPE-PEG-FSH, and 8-12 parts Panax notoginseng saponin Ft1.

[0009] Preferably, the notoginsenoside Ft1 liposome nanoparticles of the present invention are made from the following components in parts by weight: 40 parts soybean lecithin, 2.5 parts cholesterol, 2.5 parts DSPE-PEG-FSH, and 10 parts notoginsenoside Ft1.

[0010] The specific steps of the preparation method of the notoginsenoside Ft1 liposome nanoparticles of the present invention are as follows: weigh soybean lecithin, cholesterol, DSPE-PEG-FSH and notoginsenoside Ft1 according to the formula composition, dissolve them in chloroform, and evaporate them under reduced pressure in a container to form a film; add deionized water solution for hydration, treat with ultrasound and liposome extruder, dialyze with a nanodialysis device, and make up the volume to obtain the nanoparticles.

[0011] Preferably, the specific steps of the preparation method of the notoginsenoside Ft1 liposome nanoparticles of the present invention are as follows: take 40 mg of soybean lecithin, 2.5 mg of cholesterol, 2.5 mg of DSPE-PEG-FSH, and 10 mg of notoginsenoside Ft1, dissolve them in 3 mL of chloroform, and form a film under reduced pressure in a container; add deionized water solution for hydration, treat with ultrasound and a liposome extruder, dialyze with a nanodialysis device, and make up to 5 mL to obtain the nanoparticles; wherein, the liposome extruder is equipped with a 100 nm polycarbonate membrane, and the nanodialysis device is equipped with a 30 nm polycarbonate membrane.

[0012] The present invention relates to the application of Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of a drug for the prevention or treatment of cisplatin-induced ovarian reserve deficiency. The drug uses Panax notoginseng saponin Ft1 liposome nanoparticles as the main active ingredient and adds pharmaceutically acceptable excipients to prepare a pharmaceutically acceptable formulation.

[0013] Preferably, in the application of the notoginsenoside Ft1 liposome nanoparticles of the present invention in the preparation of a drug for the prevention or treatment of cisplatin-induced ovarian reserve deficiency, the dosage form of the drug is an injection.

[0014] Preferably, the application of the Panax notoginseng saponin Ft1 liposome nanoparticles of the present invention in the preparation of drugs that enhance the quantity and quality of oocytes.

[0015] Preferably, the application of the Panax notoginseng saponin Ft1 liposome nanoparticles of the present invention in the preparation of drugs for the prevention or treatment of premature ovarian failure.

[0016] Preferably, the application of the Panax notoginseng saponin Ft1 liposome nanoparticles of the present invention in the preparation of drugs that inhibit ovarian stromal cell fibrosis.

[0017] Preferably, the application of the Panax notoginseng saponin Ft1 liposome nanoparticles of the present invention in the preparation of drugs to enhance fertility.

[0018] The beneficial effects of this invention are: 1. This invention successfully prepared FSH-modified Panax notoginseng saponin Ft1 ovarian-targeting liposomes (FSH-Lipo@Ft1). The liposome nanoparticles have a spherical vesicle structure, uniform particle size, concentrated distribution, and good colloidal stability and structural integrity.

[0019] 2. In vivo pharmacodynamic experiments have confirmed that the Panax notoginseng saponin Ft1 liposome nanoparticles provided in this invention have the following effects: They can effectively alleviate weight loss in mice induced by cisplatin chemotherapy, and restore ovarian volume and quality (FSH-Lipo@Ft1 can restore 40%-58% of mouse weight); they can repair ovarian tissue structure and maintain follicular reserve; they can inhibit ovarian stromal cell fibrosis (FSH-Lipo@Ft1 can reduce the fluorescence intensity of fibrosis markers Sirius red, α-SMA, and Collagen 3 by 54%-72%), and reverse ovarian cell senescence; they can enhance ovarian responsiveness and improve superovulation efficiency (FSH-Lipo@Ft1 can increase ovulation efficiency by 66%); they can significantly increase the number and quality of oocytes (FSH-Lipo@Ft1 can reduce cytoplasmic maturity defects from 73.1% to 12.2%); and they can salvage mouse fertility. This invention can achieve the dual goals of targeted drug delivery and functional recovery in cisplatin-induced POF, providing a superior nanotherapy strategy for ovarian protection and possessing great development and application potential. Attached Figure Description

[0020] Figure 1 The results of the characterization experiments on liposome nanoparticles are shown in the figure (A is the TEM image; B is the hydrodynamic particle size distribution of liposome nanoparticles; C is the UV and fluorescence spectra; D is the Fourier transform infrared spectrum; E is the elemental mapping of FSH-Lipo@Ft1). Figure 2 The results of the biosafety experiments on Ft1 nanomaterials are shown in the figure (A is the hemolysis test result; B is the quantitative analysis result of the hemolysis test; C is the HE staining result of the heart, liver, spleen, lung, kidney and ovary; D is the statistical result of serum urea nitrogen, creatinine, AST and ALT detected by ELISA). Figure 3 The results of an in vivo efficacy study of FSH-Lipo@Ft1 in treating DDP-induced premature ovarian failure are shown in the figure. (A shows the morphological observation of oocytes in each group; B shows the Ovastacin fluorescence staining of oocytes, with green representing Ovastacin and blue representing DAPI; C shows the immunofluorescence staining of oocyte spindles, with red representing α-Tubulin and blue representing DAPI; D shows the fertility assessment results of mice after 8 months of continuous mating experiments in each treatment group; E shows the macroscopic morphological observation of ovaries in each treatment group; F shows the HE staining results of ovarian tissue in each treatment group; G shows the Sirius red staining results of ovarian tissue in each treatment group; HI shows the fluorescence intensity of α-SMA and Collagen3 detected by immunofluorescence in each treatment group). Detailed Implementation

[0021] The technical solution of the present invention will be described in detail below with reference to specific embodiments. The following embodiments are for explanation and illustration only, and do not constitute a limitation on the technical solution of the present invention.

[0022] Example 1

[0023] The liposome nanoparticles of Panax notoginseng saponin Ft1 are made from the following components: 40 mg soybean lecithin, 2.5 mg cholesterol, 2.5 mg DSPE-PEG-FSH, and 10 mg Panax notoginseng saponin Ft1.

[0024] Example 2

[0025] The liposome nanoparticles of Panax notoginseng saponin Ft1 are made from the following components: 35 mg soybean lecithin, 2.0 mg cholesterol, 2.0 mg DSPE-PEG-FSH, and 8 mg Panax notoginseng saponin Ft1.

[0026] Example 3

[0027] The liposome nanoparticles of Panax notoginseng saponin Ft1 are made from the following components: 45 mg soybean lecithin, 3 mg cholesterol, 3 mg DSPE-PEG-FSH, and 12 mg Panax notoginseng saponin Ft1.

[0028] Example 4

[0029] The preparation method of Panax notoginseng saponin Ft1 liposome nanoparticles is as follows: According to the formulation in Examples 1-3, weigh soybean lecithin, cholesterol, DSPE-PEG-FSH and Panax notoginseng saponin Ft1, dissolve them in 3 mL of chloroform, and form a film under reduced pressure in a container; add deionized water for hydration, treat with ultrasound and a liposome extruder (equipped with a 100 nm polycarbonate membrane), dialyze with a nanodialysis device (equipped with a 30 nm polycarbonate membrane), and make up to 5 mL to obtain the final product.

[0030] Example 5

[0031] Using Panax notoginseng saponin Ft1 liposome nanoparticles as raw material, deionized water was added to prepare an injection solution with a concentration of 20 μM.

[0032] Example 6

[0033] The Panax notoginseng saponin Ft1 liposome nanoparticle injection prepared in Example 5 was used to prevent or treat cisplatin-induced ovarian reserve decline, increase the number and quality of oocytes, prevent or treat premature ovarian failure, inhibit ovarian stromal cell fibrosis, or improve fertility, with a dosage of 10 mg / kg.

[0034] To further verify the reliability of the present invention, the inventors conducted a series of experiments, as follows: 1. Instruments and Materials The main instruments are listed in Table 1, and the main materials are listed in Table 2.

[0035] ; .

[0036] 2. Preparation of Panax notoginseng saponin Ft1 liposome nanoparticles (1) Preparation of blank liposome nanoparticles (Lipo) Take 50 mg of soybean lecithin, 2.5 mg of cholesterol, and 2.5 mg of DSPE-PEG, dissolve them in 3 mL of chloroform, and evaporate them under reduced pressure in a sample vial to form a membrane; add deionized water for hydration, treat with ultrasound and a liposome extruder (polycarbonate membrane, 100 nm pore size), dialyze with a nanodialysis device (polycarbonate membrane, 30 nm pore size), and bring the volume to 5 mL.

[0037] (2) Preparation of Panax notoginseng saponin Ft1 and fluorescent Cy5.5 liposome nanoparticles (Lipo@Ft1) Take 40 mg of soybean lecithin, 2.5 mg of cholesterol, 2.5 mg of DSPE-PEG, 1.5 mg of Cy5.5, and 10 mg of Panax notoginseng saponin Ft1, dissolve them in 3 mL of chloroform, and evaporate them under reduced pressure in a sample vial to form a membrane; add deionized water for hydration, treat with ultrasound and liposome extruder (polycarbonate membrane, 100 nm pore size), dialyze with a nanodialysis device (polycarbonate membrane, 30 nm pore size), and bring the volume to 5 mL.

[0038] (3) Preparation of liposome nanoparticles loaded with Panax notoginseng saponin Ft1 and fluorescent Cy5.5 and modified with FSH-targeting peptide (FSH-Lipo@Ft1) Take 40 mg of soybean lecithin, 2.5 mg of cholesterol, 2.5 mg of DSPE-PEG-FSH, 1.5 mg of Cy5.5, and 10 mg of Panax notoginseng saponin Ft1, dissolve them in 3 mL of chloroform, and evaporate them under reduced pressure in a sample vial to form a membrane; add deionized water for hydration, treat with ultrasound and liposome extruder (polycarbonate membrane, 100 nm pore size), dialyze with a nanodialysis device (polycarbonate membrane, 30 nm pore size), and bring the volume to 5 mL.

[0039] 3. Characterization of Panax notoginseng saponin Ft1 liposome nanoparticles To achieve precise targeted delivery of Ft1 to the ovary, this study successfully constructed a follicle-stimulating hormone (FSH) modified liposome nanocarrier (FSH-Lipo@Ft1), and systematically evaluated its physicochemical properties and targeting ability. The results are shown in Table 1.

[0040] Transmission electron microscopy (TEM) results showed that Lipo, Lipo@Ft1, and FSH-Lipo@Ft1 all exhibited uniformly shaped spherical vesicle structures. Figure 1 A); Dynamic light scattering (DLS) analysis showed that it had a narrow particle size distribution (polydispersity index PDI < 0.2) and no obvious aggregation ( Figure 1 B); UV-Vis absorption spectroscopy confirmed the successful labeling of Cy5.5 in liposomes ( Figure 1 C); Fourier transform infrared (FTIR) spectroscopy shows that FSH-Lipo@Ft1 simultaneously possesses characteristic absorption peaks of Lipo, Ft1, and FSH, and is located at 1093 cm⁻¹. -1 The characteristic peaks observed at this location confirm the successful coupling of liposomes with FSH. Figure 1 D); Elemental mapping analysis further verified the uniform distribution of its characteristic elements such as C, N, and P ( Figure 1 E).

[0041] 4. In vivo safety studies of FSH-Lipo@Ft1 and Lipo@Ft1 4.1 Experimental Methods (1) Hemolysis test Mice were anesthetized by enucleation, and blood was collected in centrifuge tubes containing anticoagulant and gently mixed. The mixture was immediately centrifuged at 3000 rpm for 15 min, and the supernatant plasma and leukocyte layer were carefully discarded. An appropriate amount of 0.9% physiological saline was added to the erythrocyte pellet, and the cells were gently resuspended. The pellet was then centrifuged again at 3000 rpm for 10 min, and the supernatant was discarded. This washing process was repeated 2-3 times until the supernatant was clear and colorless. After the final centrifugation, the supernatant was discarded, and the erythrocytes were resuspended in 0.9% physiological saline to prepare a 5% (v / v) erythrocyte suspension (e.g., 50 μL hematocrit + 950 μL physiological saline).

[0042] Prepare test material solutions with different concentration gradients (dissolved or diluted with 0.9% physiological saline). Take 1.5 mL centrifuge tubes, add 1 mL of 5% erythrocyte suspension to each tube, centrifuge at 3000 rpm for 10 min, and carefully discard the supernatant after a dense erythrocyte layer is formed. Add 1 mL of test material solution of different concentrations to the erythrocyte pellet in each tube, ensuring sufficient contact between the erythrocytes and the test material. Simultaneously set up a negative control (1 mL of 0.9% physiological saline) and a positive control (1 mL of deionized water). Incubate all centrifuge tubes in a 37℃ water bath for 2 h, avoiding shaking during this period.

[0043] Centrifuge the centrifuge tubes at 10,000 rpm for 20 minutes to precipitate undissolved red blood cells. Prepare a folded piece of white paper and attach double-sided tape along the fold. After centrifugation, quickly attach the centrifuge tubes side by side in the same direction to the double-sided tape, visually observe and photograph the color of the supernatant in each tube.

[0044] Carefully pipette 150 μL of the supernatant from each tube and add it to a 96-well plate. Measure the absorbance (OD value) of each well using a microplate reader at 540 nm. Calculate the hemolysis rate: Hemolysis rate (%) = [(OD value of experimental group – OD value of negative control group) / (OD value of positive control group – OD value of negative control group)] × 100%.

[0045] (2) Liver and kidney function tests Blood was collected from the ocular veins of anesthetized mice and allowed to stand at room temperature for approximately 30 minutes to allow for complete coagulation. The serum was then separated and collected by centrifugation at 5000 rpm for 15 minutes. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (Cr), and blood urea nitrogen (BUN) levels were measured according to the manufacturer's instructions using the ALT kit, AST kit, creatinine (Cr) assay kit, and BUN test kit, respectively.

[0046] (3) HE staining The paraffin sections were treated sequentially in the following solutions: xylene: twice, 10 min each time; 100% ethanol: twice, 4 min each time; 95% ethanol: once, 4 min; 80% ethanol: once, 4 min; 70% ethanol: once, 4 min; and rinsed three times in PBS buffer, 5 min each time.

[0047] Immerse the slides in antigen retrieval solution and place them in an autoclave for retrieval for 25 minutes. After allowing them to cool naturally to room temperature, replace the solution with PBS buffer and wash the slides four times on a shaker at 90 rpm for 4 minutes each time.

[0048] Hematoxylin staining: Pour off excess PBS from the slide, immerse it in hematoxylin staining solution for 10-60 seconds, remove it quickly and rinse it with running tap water.

[0049] Blueing: Immerse the slides in running tap water for about 20 minutes. During this time, observe the staining under an optical microscope until the cell nuclei turn pale blue and the non-nucleus areas remain unstained.

[0050] Eosin staining: Pour off excess water and immerse the sections in eosin staining solution for 10-20 seconds. During the staining process, gently shake the sections up and down continuously to ensure even staining. After staining, rinse thoroughly with running tap water and observe the eosin staining effect under a microscope.

[0051] Dehydration and clearing: The sections were quickly immersed in the following solutions in sequence: 70% ethanol: shake for 3 seconds; 80% ethanol: shake for 3 seconds; 90% ethanol: shake for 5 seconds; 100% ethanol: twice, shake for 5 seconds each time; xylene: twice, shake for 3 minutes each time.

[0052] Neutral resin mounting: Place a small drop of neutral resin in the center of a coverslip and quickly cover the tissue section. Place the mounted slide in a fume hood to air dry. After the sections are completely mounted and dry, observe and acquire images.

[0053] 4.2 Results and Analysis See results Figure 2Hemolysis experiments showed that, within a concentration range of 100 μg / mL to 5 mg / mL, the hemolysis rates of both Lipo@FT1 and FSH-Lipo@FT1 were well below the safety threshold of 5%, indicating good blood compatibility. Figure 2 A-2B); HE staining results showed that, compared with the control group, no obvious pathological damage or inflammatory infiltration was observed in the major organs (heart, liver, spleen, lung, kidney, and ovary) of mice treated with Lipo@FT1 and FSH-Lipo@FT1. Figure 2 C); Serum biochemical analysis 14 days after injection showed no significant differences in renal function indicators (creatinine, blood urea nitrogen) and liver function indicators (alanine aminotransferase, aspartate aminotransferase) levels among the groups. Figure 2 (D) further confirms that FSH-Lipo@FT1 has no significant adverse effects on liver and kidney function and tissue morphology.

[0054] 5. In vivo efficacy study of FSH-Lipo@Ft1 in treating DDP-induced premature ovarian failure 5.1 Experimental Methods (1) Construction of mouse rescue models of Lipo@Ft1 and FSH-Lipo@Ft1 Eight-week-old female KM mice of similar weight were randomly divided into four groups: Control group, DDP group, DDP+Lipo@Ft1 group, and DDP+FSH-Lipo@Ft1 group. The DDP group received intraperitoneal injections of cisplatin 2 mg / kg for 14 consecutive days; the DDP+Lipo@Ft1 group received cisplatin 2 mg / kg concurrently with intraperitoneal injections of Lipo@Ft1 10 mg / kg for 14 consecutive days; the DDP+FSH-Lipo@Ft1 group received cisplatin 2 mg / kg concurrently with FSH-Lipo@Ft1 10 mg / kg for 14 consecutive days; the control group received an equal volume of physiological saline intraperitoneally during the same period. On day 15, all mice were intraperitoneally injected with pregnant mare serum gonadotropin (PMSG) to simulate FSH effects, followed by injection of human chorionic gonadotropin (hCG) 48 hours later to simulate the LH surge. The animals were sacrificed 15 hours later for tissue collection. Throughout the drug administration period, the dynamic changes in the body weight of mice in each group were recorded. After sacrifice, the bilateral ovaries were separated, weighed, and their appearance was recorded by photographing under a stereomicroscope.

[0055] (2) Immunofluorescence staining experiment of oocytes For oocyte collection, after euthanasia, the ampulla segments of both fallopian tubes were quickly removed and placed in pre-cooled PBS. Under a stereomicroscope, the fallopian tube wall was gently punctured with a sterile needle to release the cumulus-oocyte complex into the buffer solution. The oocyte mass was then transferred to trypsin digestion solution and observed under a microscope in real time. When the granulosa cells surrounding the oocytes were largely dissociated, the digestion was immediately terminated by aspirating the oocytes into serum-containing culture medium using an oral pipette. After washing three times with fresh culture medium to remove residual granulosa cells, the oocytes were fixed in 4% paraformaldehyde. After natural settling, they were observed, counted, and photographed under an inverted microscope.

[0056] For immunofluorescence staining, 100 μL of permeabilization working solution was first added to a new culture dish, and the fixed oocytes were transferred into it and incubated at room temperature for 30 min to complete the permeabilization process. Then, the oocytes were transferred to 100 μL of blocking solution and blocked at room temperature for 1 h. After blocking, 100 μL of diluted primary antibody working solution was added, and the cells were incubated overnight at 4 ℃. The next day, the oocytes were transferred to washing buffer, gently pipetted, and the buffer was replaced with fresh buffer. This washing process was repeated three times, 10 min each time. Then, 100 μL of fluorescent secondary antibody working solution was prepared using the blocking solution and incubated at room temperature in the dark for 2 h. The cells were then washed four times with washing buffer, 10 min each time, to ensure complete removal of the secondary antibody before slide preparation. A clean slide was taken, and the observation area was marked with a marker. Oocytes were dropped into the circle, and lanolin was applied to the four corners of the circle for support. A coverslip was gently placed on top to prevent cell rupture due to pressure. Finally, images were acquired and fluorescence signals were analyzed using a laser confocal microscope.

[0057] 5.2 Results and Analysis Results of an in vivo efficacy study of FSH-Lipo@Ft1 in treating DDP-induced premature ovarian failure are shown below. Figure 3 .

[0058] Superovulation experiments showed cisplatin-induced oocyte exhaustion and abnormalities (spindle / chromosomal alignment abnormalities). The results indicated that FSH-Lipo@Ft1 was superior to Lipo@Ft1 in restoring normal oocyte number and morphology (reducing spindle defects and chromosome dispersion). FSH-Lipo@Ft1 increased ovulation efficiency in mice by 66% and reduced cytoplasmic maturity (lectin / oocyte antigen) defects from 73.1% (cisplatin) to 12.2%. Figure 3 AC).

[0059] Depend on Figure 3 As shown in D, FSH-Lipo@Ft1 showed the best fertility recovery effect, and compared with the Lipo@Ft1 group, it was more effective in restoring the litter size to normal.

[0060] Cisplatin can lead to a reduction in ovarian volume and the number of follicles, while both groups of drug-loaded nanoparticles can reverse these damages, with FSH-Lipo@Ft1 showing significantly higher efficacy. Figure 3 (E-3F). Furthermore, FSH-Lipo@Ft1 effectively alleviated the weight loss induced by cisplatin chemotherapy in mice, restoring 40%-58% of the mice's body weight.

[0061] Depend on Figure 3 H-3I analysis revealed that the fluorescence intensity of fibrosis markers (Sirius Red, α-SMA, Collagen 3) decreased by 54%-72%.

[0062] In summary, the results of this study indicate that the Panax notoginseng saponin Ft1 liposome nanoparticles effectively alleviated the weight loss induced by cisplatin chemotherapy in mice, restored ovarian volume and quality, increased follicle reserve, improved oocyte quality, and significantly protected ovarian tissue structure and overall fertility. This study achieves the dual goals of targeted drug delivery and functional recovery in cisplatin-induced POF, providing a superior nanotherapy strategy for ovarian protection.

[0063] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A liposome nanoparticle containing Panax notoginseng saponin Ft1, characterized in that, The notoginsenoside Ft1 liposome nanoparticles are made from the following components in parts by weight: 35-45 parts soybean lecithin, 2-3 parts cholesterol, 2-3 parts DSPE-PEG-FSH, and 8-12 parts notoginsenoside Ft1.

2. The Panax notoginseng saponin Ft1 liposome nanoparticles according to claim 1, characterized in that, The notoginsenoside Ft1 liposome nanoparticles are made from the following components in parts by weight: 40 parts soybean lecithin, 2.5 parts cholesterol, 2.5 parts DSPE-PEG-FSH, and 10 parts notoginsenoside Ft1.

3. The method for preparing Panax notoginseng saponin Ft1 liposome nanoparticles as described in claim 1, characterized in that, The specific steps are as follows: Weigh soybean lecithin, cholesterol, DSPE-PEG-FSH and Panax notoginseng saponin Ft1 according to the formula composition, dissolve them in chloroform, and evaporate them under reduced pressure in a container to form a film; add deionized water solution for hydration, treat with ultrasound and liposome extruder, dialyze with a nanodialysis device, and make up to a fixed volume to obtain the final product.

4. The method for preparing Panax notoginseng saponin Ft1 liposome nanoparticles according to claim 3, characterized in that, The specific steps are as follows: Take 40 mg of soybean lecithin, 2.5 mg of cholesterol, 2.5 mg of DSPE-PEG-FSH, and 10 mg of Panax notoginseng saponin Ft1, dissolve them in 3 mL of chloroform, and form a film under reduced pressure in a container; add deionized water solution for hydration, treat with ultrasound and a liposome extruder, dialyze with a nanodialysis device, and make up to 5 mL to obtain the final product; wherein, the liposome extruder is equipped with a 100 nm polycarbonate membrane, and the nanodialysis device is equipped with a 30 nm polycarbonate membrane.

5. The application of the Panax notoginseng saponin Ft1 liposome nanoparticles as described in claim 1 in the preparation of drugs for the prevention or treatment of cisplatin-induced ovarian reserve deficiency, characterized in that... The drug uses Panax notoginseng saponin Ft1 liposome nanoparticles as the main active ingredient, and is prepared into a pharmaceutically acceptable formulation with the addition of pharmaceutically acceptable excipients.

6. The application according to claim 5, characterized in that, The drug for preventing or treating cisplatin-induced ovarian reserve decline is in the form of an injection.

7. The application according to claim 5, characterized in that, Application of the Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of drugs that enhance the quantity and quality of oocytes.

8. The application according to claim 5, characterized in that, Application of the Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of drugs for the prevention or treatment of premature ovarian failure.

9. The application according to claim 5, characterized in that, Application of the Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of drugs that inhibit ovarian stromal cell fibrosis.

10. The application according to claim 5, characterized in that, Application of the Panax notoginseng saponin Ft1 liposome nanoparticles in the preparation of drugs to enhance fertility.

Images