An ovary maintenance bacteriostatic gel and a preparation method and application thereof

Abstract

The application relates to the field of bacteriostatic gels, in particular to an ovary maintenance bacteriostatic gel as well as a preparation method and application thereof. The ovary maintenance bacteriostatic gel comprises A component, B component and C component; the A component comprises the following components in parts by weight: deionized water 64-84 parts, preservative synergist 0.05-0.15 parts, Carbopol U-30 0.6-0.9 parts, super high molecular weight hyaluronic acid 0.06-0.12 parts, hydrogel 3-7 parts, glycerol 3-6 parts and propylene glycol 3-6 parts; the B component comprises the following components in parts by weight: selective estrogen receptor modulator 3-7 parts and radix viticis trifolia extract 0.5-2.5 parts; and the C component comprises the following components in parts by weight: silver ion solution 0.1-0.3 parts, preservative 0.3-0.6 parts, Tween-20 0.05-0.15 parts and triethanolamine 0.05-0.25 parts. The gel obtained by the application can not only play a bacteriostatic role when applied to ovaries, but also can effectively improve premature ovarian failure.

Description

Technical Field

[0001] This application relates to the field of antibacterial gels, and in particular to an ovarian maintenance antibacterial gel, its preparation method, and its application. Background Technology

[0002] The ovary is an important endocrine organ in women, responsible for ovulation and the secretion of sex hormones (such as estrogen and progesterone). Its health is closely related to a woman's reproductive capacity, overall health, and aging process.

[0003] The ovary is divided into inner and outer surfaces, upper and lower ends, and anterior and posterior borders. The inner surface of the ovary faces the pelvic cavity and is often adjacent to the ileum, hence its name "intestinal surface." The outer surface is in contact with the pelvic sidewall. The main functions of the ovary are to produce eggs, steroid hormones, and progesterone. The size of the ovary is related to age and the egg-producing period. After the age of thirty, women enter a period of ovarian function decline. Premature ovarian failure can cause a series of hormonal changes in the body, leading to symptoms such as irregular menstruation and irritability.

[0004] Premature ovarian failure (POF) refers to the decline in ovarian function in women before the age of 40. Symptoms include menstrual irregularities (amenorrhea or oligomenorrhea), elevated gonadotropin levels (FSH > 25 U / L), decreased estrogen levels, and symptoms such as hot flashes, night sweats, and infertility. It not only affects fertility but may also increase long-term health risks such as osteoporosis and cardiovascular disease.

[0005] Therefore, improving premature ovarian failure is extremely important. Existing methods for improving premature ovarian failure include hormone therapy or hormone replacement therapy, non-hormonal drug therapy, traditional Chinese medicine, and immunotherapy. Among these, hormone therapy has significant side effects, traditional Chinese medicine is laborious, time-consuming, and has weak effects, immunotherapy is expensive, and non-hormonal drug therapy generally uses selective serotonin reuptake inhibitors, which are not very effective. Therefore, it is essential to develop a product that is easy to use, has few side effects, and is effective. Summary of the Invention

[0006] In order to reduce side effects when effectively maintaining the ovaries, this application provides an ovarian maintenance antibacterial gel, its preparation method and application.

[0007] In the first aspect, this application provides an ovarian maintenance antibacterial gel, which adopts the following technical solution.

[0008] An ovarian maintenance antibacterial gel, comprising component A, component B and component C;

[0009] Component A comprises the following raw materials in parts by weight: 64-84 parts deionized water, 0.05-0.15 parts preservative synergist, 0.6-0.9 parts carbomer U-30, 0.06-0.12 parts ultra-high molecular weight hyaluronic acid, 3-7 parts hydrogel, 3-6 parts glycerin, and 3-6 parts propylene glycol;

[0010] Component B comprises the following raw materials in parts by weight: 3-7 parts of selective estrogen receptor modulator and 0.5-2.5 parts of *Solanum nigrum* extract;

[0011] The C component comprises the following raw materials in parts by weight: 0.1-0.3 parts silver ion solution, 0.3-0.6 parts preservative, 0.05-0.15 parts Tween-20, and 0.05-0.25 parts triethanolamine.

[0012] By adopting the above technical solutions, selective estrogen receptor modulators can bidirectionally regulate and promote the restoration of normal gonadal axis function, deeply repair, regulate internal circulation, restore vitality, add ovarian nutrients, deeply nourish and repair the ovaries, prevent premature ovarian failure, and alleviate ovarian aging.

[0013] The extract of Sufanghua (a type of flower) contains a variety of functional components, commonly including phenolic compounds, terpenoids, sterols and flavonoids, and volatile components.

[0014] Ovarian dysfunction is closely related to oxidative stress. Excessive free radicals can damage oocyte DNA, mitochondrial function, and theca cells, leading to accelerated follicular apoptosis. The polyphenols in *Solanum nigrum* extract neutralize free radicals through a direct electron transfer mechanism, while simultaneously upregulating the activity of antioxidant enzymes (such as SOD and GSH-Px), enhancing the ovarian tissue's antioxidant defense system, scavenging free radicals such as superoxide anions and hydroxyl radicals, reducing oxidative damage to ovarian tissue, and delaying cell aging.

[0015] Terpenoids can inhibit the activation of the NF-κB signaling pathway and reduce the transcription of inflammatory factors; rutin can block the synthesis of pro-inflammatory mediators such as prostaglandins and leukotrienes by inhibiting the activity of cyclooxygenase and lipoxygenase; and the extract of *Solanum nigrum* may inhibit fibroblast activation, reduce the degree of tissue fibrosis, inhibit local inflammatory response in the ovary, reduce the release of pro-inflammatory factors, and improve the inflammatory pathological state of follicle development by downregulating the expression of transforming growth factor β1.

[0016] Flavonoids have structures similar to estradiol and can weakly competitively bind to estrogen receptors, exerting a bidirectional regulatory effect. They mimic the effects of estrogen when it is insufficient, but antagonize it when it is excessive. They may indirectly improve the symptoms of ovarian insufficiency caused by low estrogen levels through phytoestrogen activity or regulation of hormone metabolism enzymes.

[0017] The extract of Sufanghua can also be absorbed through the skin and act on peripheral vascular β2 adrenergic receptors, indirectly improving microcirculation, promoting local vasodilation in the ovary, increasing blood flow, and optimizing the nutritional supply for follicle development.

[0018] Furthermore, the selective estrogen receptor modulator includes at least two of the following: propylene glycol / pueraria lobata extract, Cuscuta chinensis seed extract, Spatholobus suberectus extract, and flaxseed oil / Dioscorea opposita root extract, as well as tocopheryl acetate, thiamine hydrochloride, pyridoxine, oregano leaf extract, and lactic acid bacteria fermentation products.

[0019] Furthermore, based on the total weight of the selective estrogen receptor modulator, it comprises the following raw materials in parts by weight: 1-3 parts of propylene glycol / kudzu root extract, 0.5-2 parts of Cuscuta chinensis seed extract, 0.5-1.5 parts of Spatholobus suberectus extract, 1-2.5 parts of linseed oil / Dioscorea opposita root extract, 0.1-0.5 parts of tocopheryl acetate, 0.05-0.2 parts of thiamine hydrochloride, 0.05-0.2 parts of pyridoxine, 0.3-1 parts of oregano leaf extract, and 2-5 parts of lactic acid bacteria fermentation product.

[0020] By adopting the above technical solutions, puerarin and soy isoflavones have structures similar to estradiol and can weakly competitively bind to estrogen receptors to exert a bidirectional regulatory effect. When estrogen levels are insufficient in the body, they mimic the effects of estrogen, promote the proliferation of follicular granulosa cells, and upregulate aromatase activity to increase estrogen synthesis. When estrogen levels are too high, they competitively occupy receptor sites, inhibit excessive secretion of luteinizing hormone, and reduce the hyperandrogen state of the ovaries.

[0021] Flavonoids in Cuscuta chinensis seed extract can activate the PI3K / Akt signaling pathway, promote the transformation of primordial follicles into primary follicles, and delay follicle pool depletion; sterols can reduce the conversion of testosterone to dihydrotestosterone by inhibiting reductase, improve hyperandrogenemia, and restore ovulation function; polysaccharide components can downregulate the expression of transforming growth factor type I collagen, inhibit fibroblast activation, and prevent the progression of inflammatory ovarian fibrosis.

[0022] The phenolic acids in chicken blood vine extract promote NO production by activating endothelial nitric oxide synthase, dilate ovarian vascular smooth muscle, increase follicular blood perfusion, and optimize oocyte nutrition supply; flavonoids inhibit the release of platelet aggregation factor, reduce blood viscosity, and improve microcirculatory disorders in patients with "qi stagnation and blood stasis type" ovarian hypofunction; sterols bind weakly to estrogen receptors and synergistically enhance hormone regulation with kudzu root extract.

[0023] Diosgenin, a component of yam root extract, is a natural precursor for the synthesis of progesterone and estrogen. It can be absorbed through the skin and metabolized in the liver to pregnenolone, which is then converted into progesterone and estrogen, indirectly supplementing insufficient corpus luteum function in the ovary. Its metabolites can enhance mitochondrial membrane stability, reduce the production of reactive oxygen species during oocyte meiosis, reduce the incidence of aneuploid oocytes, and improve oocyte quality.

[0024] Terpenoids and phenolic compounds in oregano leaf extract bind to bacterial cell membrane phospholipids, increasing membrane permeability and leading to ATP leakage and intracellular enzyme inactivation; they also target the mitochondrial electron transport chain, blocking bacterial energy synthesis; and they inhibit the quorum sensing system of pathogenic bacteria such as Pseudomonas aeruginosa, enhancing antibiotic sensitivity.

[0025] Lactic acid bacteria fermentation products create an acidic environment, lowering the vaginal pH to 3.8-4.2, inhibiting the overgrowth of aerobic bacteria and fungi, and maintaining the homeostasis of the lactobacillus-dominated microecology; secreting antibiotic-like substances specifically disrupt the peptidoglycan structure of Gram-positive bacterial cell membranes, leading to cell lysis.

[0026] Tocopheryl acetate, as an antioxidant, protects the vaginal mucosal epithelial cell membrane from oxidative damage, maintains the integrity of cell tight junctions, and reduces the entry point for pathogens; it also enhances the phagocytic activity of neutrophils and macrophages, thereby improving local immune defense capabilities.

[0027] Kudzu root, yam, and dodder seed form a hormonal balancing network, suitable for different types of hormonal imbalances; chicken blood vine and propylene glycol enhance the transdermal absorption efficiency of active ingredients, especially suitable for improving ovarian blood flow during application. Oregano leaf, lactic acid bacteria, and propylene glycol form a triple defense line of "physical destruction + metabolic inhibition + microecological regulation," covering multiple pathogens including aerobic bacteria, anaerobic bacteria, and fungi; tocopheryl acetate and lactic acid bacteria metabolites can reduce inflammatory damage and promote post-infection vaginal epithelial repair.

[0028] It plays a role in ovarian maintenance and antibacterial scenarios through multiple pathways, including phytoestrogen mimicry, hormone metabolism regulation, antioxidant stress, microecological regulation, and direct pathogen inhibition.

[0029] Furthermore, the weight ratio of propylene glycol / kudzu root extract, dodder seed extract, chicken blood vine extract, and flaxseed oil / yam root extract is 3:1.2:1:2.5.

[0030] Furthermore, the preparation method of the *Solanum nigrum* extract is as follows:

[0031] 1) Raw material preparation

[0032] Dry and pulverize the plain flower;

[0033] 2) One extraction

[0034] Soak the crushed Su Fanghua for 30-35 minutes, then distill it at a pressure of 0.1-0.2 MPa and a temperature of 96-100℃. Collect the distillate after 3 hours and collect the residue. Separate the oil and water from the distillate, collect the upper volatile oil, and store it in the dark and cold.

[0035] 3) Secondary extraction

[0036] Step 2) After distillation, the residue is pressed and dehydrated, ethanol is added, and ultrasonic treatment is performed at 40-45℃. Then, it is refluxed and extracted twice at 80-83℃, 2 hours each time. After reflux extraction, it is filtered while hot, the filtrates are combined, and the residue is collected.

[0037] 4) Three extractions

[0038] In step 3), the ethanol in the filter residue is evaporated, water is added, and the mixture is stirred and extracted at 60-65°C for 4 hours. The supernatant is then collected by centrifugation.

[0039] 5) Emulsification

[0040] Combine the filtrate obtained in step 3) with the supernatant obtained in step 4), and add the volatile oil and emulsifier obtained in step 1), homogenize until the emulsion particle size is ≤5μm;

[0041] 6) Filter, concentrate, and dry to obtain Sufanghua extract.

[0042] Furthermore, in step 5), the weight ratio of emulsifier to volatile oil is 1:1.

[0043] By adopting the above technical solution and using polar step extraction, the problem of component mutual damage was solved.

[0044] Furthermore, the corrosion inhibitor is EDTA-2Na.

[0045] Furthermore, the hydrogel is a Polymatrix7 hydrogel.

[0046] Secondly, this application provides a method for preparing an ovarian maintenance antibacterial gel, which adopts the following technical solution.

[0047] A method for preparing an ovarian maintenance and antibacterial gel includes the following steps:

[0048] S1. Mix all the materials in phase A, and heat to 85-95℃ while stirring. After stirring until all materials are completely melted, start cooling.

[0049] S2. When the temperature drops to 40-50℃, add phases B and C and stir for 30 minutes;

[0050] S3. Stop stirring, take samples for testing, and filter out the material after all physical and chemical indicators have passed the test.

[0051] By adopting the above technical solution, the preparation method is simple and quick, has no special requirements for equipment, and is suitable for industrial production.

[0052] Thirdly, this application provides an application of an ovarian maintenance antibacterial gel, employing the following technical solution.

[0053] An application of an ovarian maintenance antibacterial gel, which is applied to the vagina by means of a paste.

[0054] By adopting the above technical solution, it is easy to use and does not require oral administration.

[0055] In summary, this application has the following beneficial effects:

[0056] Selective estrogen receptor modulators and Sufanghua extract work synergistically to increase the growth rate of FSH and LH. When applied to premature ovarian failure, the growth rate of FSH can reach 23.0-28.3%, and the growth rate of LH can reach 29.7-34.0%, effectively improving premature ovarian failure. Detailed Implementation

[0057] The present application will be further described in detail below with reference to the embodiments.

[0058] Example of raw material and intermediate preparation

[0059] raw material

[0060] All raw materials used in the embodiments of this application are commercially available.

[0061] Ultra-high molecular weight hyaluronic acid, molecular weight, 2.2 million DA;

[0062] The silver ion solution contains 0.5% silver powder by volume and 99.5% water by volume.

[0063] Preservatives, preservative CAP;

[0064] Preservative enhancer, EDTA-2Na;

[0065] Hydrogel, Polymatrix7 hydrogel.

[0066] Preparation Example

[0067] Preparation Example 1

[0068] A type of herb extract, the preparation method of which is as follows:

[0069] 1) Raw material preparation

[0070] Collect Tibetan Sufang flowers;

[0071] Graded drying:

[0072] Flowers: Dry in 40℃ hot air circulation for 4 hours → turn over → dry for another 2 hours until moisture content ≤7%;

[0073] Young leaves: Dry at 50℃ for 6 hours until moisture content is ≤8% to avoid chlorophyll degradation;

[0074] Grinding and sterilization:

[0075] Crusher parameters: rotation speed 2800rpm, screen aperture 0.8mm (24 mesh), 80% of the output particle size passes through a 60-mesh sieve;

[0076] Cobalt-60 irradiation sterilization: dose 8 kGy, kills insect eggs and microorganisms;

[0077] 2) One extraction

[0078] The crushed Su Fanghua was soaked in pure water at a ratio of 1:8 (1kg raw material: 8L water) for 30 minutes. The mixture was then distilled at a pressure of 0.15MPa and a temperature of 98℃. The distillate was collected after 3 hours, and the residue was also collected. The distillate was then separated into oil and water, and the upper volatile oil was collected and stored in the dark.

[0079] 3) Secondary extraction

[0080] Step 2) After distillation, the residue is pressed and dehydrated, then 70% ethanol is added at a material-to-liquid ratio of 1:15 (1kg raw material: 15L ethanol). The residue is ultrasonically treated at 40℃ for 30min with an ultrasonic power density of 50W / L. Then, it is refluxed twice at 80℃ for 2 hours each time. After reflux extraction, the residue is filtered through a 200-mesh filter cloth while hot, and the filtrates are combined and the residue is collected.

[0081] 4) Three extractions

[0082] The ethanol in the filter residue obtained in step 3) is evaporated, pure water is added, the material-to-liquid ratio is 1:20 (1kg raw material: 20L water), the mixture is stirred and extracted at 60℃ for 4 hours, centrifuged at 4000rpm for 15 minutes, and the supernatant is collected.

[0083] 5) Emulsification

[0084] Combine the filtrate obtained in step 3) with the supernatant obtained in step 4), and add the volatile oil and emulsifier obtained in step 1). Homogenize until the emulsion particle size is ≤5μm; the weight ratio of emulsifier to volatile oil is 1:1.

[0085] 6) Filtration, concentration, and drying

[0086] Filtered by a 100-mesh filter, then by a 0.45μm filter, and finally retained by a 10kDa hollow fiber ultrafiltration membrane;

[0087] Vacuum concentration: at a temperature of 60℃ and a vacuum degree of -0.085MPa, concentrate to a density of 1.20g / cm³;

[0088] The extract was obtained by drying at 100℃.

[0089] Preparation Example 2

[0090] A propylene glycol / pueraria extract, the preparation method of which is as follows:

[0091] 1) Raw material preparation

[0092] Collect kudzu vine;

[0093] Wash fresh kudzu root, cut it into 3mm thick slices, and dry it with hot air at 40℃ until the moisture content is ≤10%.

[0094] It is crushed into 0.4mm particles;

[0095] 2) Propylene glycol extraction

[0096] Solvent preparation: Propylene glycol: pure water = 7:3 (v / v) mixed solvent (to reduce viscosity and improve permeability);

[0097] Dynamic countercurrent extraction: solid-liquid ratio 1:12 (g / mL), temperature 60℃, time 3 hours, stirring speed 80 rpm;

[0098] Filtration: Press filter while hot using a 200-mesh filter cloth, wash the filter residue twice with 20% solvent, and combine the filtrates;

[0099] 3) Purification

[0100] Enzymatic hydrolysis of starch: Cool the filtrate to 45℃, add α-amylase (0.1% w / w), incubate for 1 hour of enzymatic hydrolysis, and inactivate at 85℃ for 10 min;

[0101] Alcohol precipitation to remove colloids: Add 95% ethanol to a final concentration of 60% (v / v), let stand for 12 hours, centrifuge at 4000 rpm for 15 min, and collect the supernatant;

[0102] Macroporous resin enrichment: Resin type AB-8, sample loading concentration of crude drug 0.5 g / mL, resin adsorption flow rate 2 BV / h (column volume), eluent 70% ethanol, elution volume 4 BV;

[0103] 4) Propylene glycol compounding and concentration

[0104] Solvent replacement: The resin eluent was concentrated under reduced pressure at 60℃ and -0.08MPa to recover ethanol, and concentrated until no alcohol odor was detected;

[0105] Propylene glycol reconstitution: Add pharmaceutical grade propylene glycol to adjust the ratio to: kudzu root extract: propylene glycol = 1:3 (w / w), and add 0.05% ethylhexylglycerin;

[0106] Homogenization: Homogenize at 10,000 rpm for 10 min, then filter through a 0.22 μm microporous membrane for sterilization to obtain propylene glycol / pueraria extract.

[0107] Preparation Example 3

[0108] A method for preparing a seed extract of Cuscuta chinensis from southern China:

[0109] 1) Raw material preparation

[0110] Select mature seeds of Cuscuta chinensis from the south;

[0111] Processing to remove toxins: Mix seeds with 2% salt solution (w / w), stir-fry over low heat until slightly puffed up, and sift to remove salt grains;

[0112] Ultrafine cell wall disruption: -196℃ liquid nitrogen flash freezing, -40℃ ultrafine pulverization, D90 particle size ≤20μm;

[0113] 2) Hierarchical extraction

[0114] Prepare the solvent: ethanol-citric acid buffer (pH 4.0) = 6:4 (v / v);

[0115] Ultrasonic-microwave synergistic extraction: material-liquid ratio 1:15 (g / mL), ultrasonic power / frequency 500W / 40kHz, microwave power / temperature 300W / 55℃, filtration to obtain filtrate and residue;

[0116] Three-stage countercurrent extraction: Three extraction tanks are connected in series, and the solvent comes into countercurrent contact with the residue for extraction;

[0117] 3) Detoxification and purification

[0118] Acid precipitation-alkali dissolution method: The filtrate obtained in step 2) is combined with the extract to form an extract. The pH is adjusted to 2.0 with 1M HCl, and the mixture is allowed to stand at 4°C for 12 hours. The precipitate is then centrifuged and discarded to obtain the supernatant.

[0119] The supernatant was adjusted to pH 8.5 with 1M NaOH, alkaloids were precipitated, and separated by a 0.45μm filter membrane;

[0120] Macroporous resin chromatography: resin type HPD-600; loading buffer pH 5.0 (citric acid buffer); adsorption flow rate 3 BV / h; elution gradient water: 30% ethanol, 70% ethanol;

[0121] 4) Concentration and Drying

[0122] Concentration under reduced pressure: elute with 70% ethanol, rotary evaporate at 50℃ and -0.09MPa to concentrate to a density of 1.25 g / cm³;

[0123] Vacuum belt drying: feed rate 10L / h, heating belt temperature 70℃ (front section), 50℃ (back section); vacuum degree -0.095MPa, moisture content ≤5%; obtained Cuscuta chinensis seed extract.

[0124] Preparation Example 4

[0125] A method for preparing a chicken blood vine extract:

[0126] 1) Raw material preparation

[0127] Select dried stems of the honeysuckle bean vine;

[0128] Enhanced efficacy through processing: Mix chicken blood vine slices with 20% rice wine (v / w), let it soak for 2 hours, then stir-fry over low heat until it turns dark red;

[0129] Ultra-low temperature cell disruption: -196℃ liquid nitrogen flash freezing, ultra-fine pulverization to D90≤30μm;

[0130] 2) Extraction

[0131] Ethanol extraction of flavonoids: solvent 70% ethanol, solid-liquid ratio 1:12 (g / mL), temperature 65℃, time 1.5 hours, ultrasonic assistance 40kHz, 300W; filtration to obtain filtrate and residue.

[0132] Phenolic acid extraction: Extraction of alcohol-extracted medicinal residue with pure water (material-to-liquid ratio 1:10) at 80℃ for 1 hour, with the addition of 0.1% ascorbic acid;

[0133] 3) Targeted purification

[0134] Composite flocculation and degumming: Combine the extracts, cool to 40℃, add 1% chitosan (w / v), stir for 30 minutes, and centrifuge at 5000 rpm for 10 minutes;

[0135] Enrichment of molecularly imprinted polymers: filler: strychnos nucifera imprinted polymer (vinylpyridine-EGDMA crosslinked), loading solution pH 5.0 (acetic acid buffer), adsorption flow rate 1.5 BV / h, eluent methanol-acetic acid (95:5).

[0136] 4) Concentration and Drying

[0137] Vacuum thin-film concentration:

[0138] Eluent → Falling film concentrator (50℃, -0.085MPa) → Concentrate to a density of 1.30g / cm³ (solid content 40%).

[0139] Freeze-spray drying: Pre-freezing temperature -40℃ (4 hours), inlet air temperature 100℃, atomization pressure 0.3MPa, particle size D50=50μm, protection 5% trehalose + 2% mannitol;

[0140] Chicken blood vine extract was obtained.

[0141] Preparation Example 5

[0142] A flaxseed oil / yam root extract, the preparation method of which is as follows:

[0143] 1) Raw material preparation

[0144] Fresh yam root slices are dried with hot air at 40℃ and then pulverized through a 40-mesh sieve.

[0145] 2) Enzymatic hydrolysis-ethanol extraction

[0146] Cellulose enzymatic hydrolysis: Raw material + water (material-to-liquid ratio 1:8), adjust pH to 5.0, add 0.2% cellulase, and enzymatically hydrolyze at 50℃ for 1 hour;

[0147] Ethanol reflux: 60% ethanol solvent, 75°C, 2 hours x 2 times, feed-to-liquid ratio 1:12

[0148] Macroporous resin purification: Resin type D101, dynamic adsorption loading flow rate 2 BV / h, water washing to remove sugar, elution with 70% ethanol (4 BV), and concentration of the eluent.

[0149] 3) Oil-extract compound emulsification

[0150] Emulsification system design:

[0151] Oil phase: 65% refined flaxseed oil, 20% ω-3 carrier diosgenin concentrate (40% solids), active ingredients (saponins ≥15%), 5% beeswax;

[0152] Aqueous phase: 8% pure water, 1.5% Tween-80, HLB=15, 0.5% ethylhexylglycerin; Emulsification process:

[0153] Oil phase mixture: flaxseed oil + yam extract + beeswax, melted and homogenized at 75℃ (5000 rpm / 5 min);

[0154] Aqueous phase dissolution: Dissolve in pure water + Tween-80 + ethylhexylglycerin by stirring at 75°C;

[0155] Primary emulsification: The aqueous phase is slowly added to the oil phase, and the mixture is subjected to high-speed shearing (10,000 rpm / 10 min) to obtain a crude emulsion;

[0156] Homogenization and refinement: High-pressure homogenizer (40MPa, 3 cycles), emulsion particle size D90≤500nm;

[0157] Degassing and filling: Vacuum degassing and nitrogen filling are used to obtain flaxseed oil / yam root extract.

[0158] Preparation Example 6

[0159] An oregano leaf extract, the preparation method of which is as follows:

[0160] 1) Raw material preparation

[0161] Greek oregano is selected, and the harvest period is before flowering.

[0162] Fresh leaves were flash-frozen in liquid nitrogen (-40℃) and then freeze-dried in vacuum (-50℃, 10Pa).

[0163] Hammer mill, for producing 2-4mm particle size fragments;

[0164] 2) Supercritical CO2 extraction

[0165] Filling density of the extraction vessel: 0.35 g / mL;

[0166] Dynamic extraction: pressure 25MPa, temperature 45℃, CO2 flow rate 40kg / h, extraction time 90 minutes; entrainer 5% ethanol (v / v).

[0167] Fractional collection: primary separator 8MPa / 30℃, secondary separator 4MPa / 25℃;

[0168] 3) Molecular distillation purification

[0169] Pretreatment: crude oil + 1% diatomaceous earth, stir and filter at 50℃;

[0170] First-stage distillation: evaporation temperature 80℃, vacuum degree 0.001Pa, feed rate 1.0mL / min, collect light fraction;

[0171] Secondary distillation: The heavy fraction (terpenes) is recycled back to the supercritical fluid extraction equipment for secondary extraction;

[0172] 4) Nanoemulsion solubilization

[0173] Formula: 10% molecularly distilled oregano oil, 15% medium-chain triglycerides (MCT), 8% Tween-80, 12% propylene glycol, 55% pure water

[0174] Process:

[0175] Oil phase mixing: oregano oil + MCT, dissolved in a 50°C water bath;

[0176] Aqueous phase dissolution: pure water + Tween-80 + propylene glycol, stirred at 50°C;

[0177] Primary emulsification: The oil phase is added to the aqueous phase and subjected to high-speed shearing (10,000 rpm / 5 min).

[0178] Homogenization: 3 cycles at 40 MPa, emulsion particle size D90≤200 nm;

[0179] Sterilization by filtration: 0.22μm PVDF filter membrane, aseptic filling, to obtain oregano leaf extract.

[0180] Preparation Example 7

[0181] A lactic acid bacteria fermentation product, the preparation method of which is as follows:

[0182] 1) Select non-GMO soybeans with a protein content ≥38%, and remove moldy and insect-infested grains;

[0183] Soak soybeans: Soybean to water ratio 1:3, add 0.5% NaHCO3 solution, soak at 30℃ for 10±0.5 hours, change the water every 2 hours, until the inner surface of the soybean is ≤1 / 2 sunken and the hardness is ≤3.5kg / cm².

[0184] Pulping: Grinding parameters: soybean-to-water ratio 1:8, colloid mill gap 50μm, two grinding cycles; Boiling conditions: dynamic boiling at 105℃ for 8 minutes, steam pressure 0.15MPa; Homogenization: pressure 35MPa, temperature 65℃, 3 cycles.

[0185] Sterilization treatment: UHT sterilization: 137℃±1℃, hold for 4 seconds, then rapidly cool to 40℃; and add 0.2% fructooligosaccharide + 0.1% yeast extract to enhance nutrition as a fermentation substrate;

[0186] 2) Mother bacteria activation: Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus jenny, Lactobacillus casei subsp. casei, Lactobacillus gasseri, Lactobacillus deutschlandiae subsp. bulgaricus, Lactobacillus helveticus, Lactococcus lactis, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus deutschlandiae, Enterococcus lactis, and Streptococcus thermophilus are cultured and activated in standard MRS medium;

[0187] 3) Inoculation and fermentation of bacterial solution:

[0188] Phase 1 (0-12h): Inoculation with *Bifidobacterium longum*, *Bifidobacterium bifidum*, and *Bifidobacterium adolescentis*; Inoculation amount: 2% mother culture, with a weight ratio of *Bifidobacterium longum*, *Bifidobacterium bifidum*, and *Bifidobacterium adolescentis* of 2:1:1; Temperature: 37℃±0.5℃, Dissolved oxygen <0.1mg / L, pH naturally decreasing; The culture solution was injected in three batches (0h, 2h, 4h); 0.05% L-cysteine ​​and 0.1% fructooligosaccharides were added to the culture medium;

[0189] Second stage (12-24h): Inoculate with *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, and *Lactobacillus casei* subsp. casei; inoculation amount: 2% mother culture, with the weight ratio of *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, and *Lactobacillus casei* subsp. casei being 3:2:1; temperature raised to 40℃±0.5℃, dissolved oxygen 0.5-1.0 mg / L, pH controlled at 5.8-6.0; 0.2% glucose added every 2 hours during the reaction; 0.01% CaCl₂ added to the culture medium;

[0190] Phase 3 (24-36h): Inoculation with *Lactobacillus delbrueckii* subsp. bulgaricus, *Lactobacillus helveticus*, and *Streptococcus thermophilus*; Inoculation amount: 2% mother culture, with a weight ratio of *Lactobacillus delbrueckii* subsp. bulgaricus, *Lactobacillus helveticus*, and *Streptococcus thermophilus* of 2:1:2; Temperature raised to 42℃±0.5℃, dissolved oxygen reduced to below 0.2 mg / L, pH allowed to decrease freely; the bacterial culture was equilibrated at 40℃ for 30 min before inoculation; 0.005% MnSO4 was added to the culture medium.

[0191] Phase 4 (36-48h): Inoculation with *Lactobacillus brevis*, *Lactobacillus janniae*, and *Lactococcus lactis*; Inoculation amount: 2% mother culture, with a weight ratio of *Lactobacillus brevis*, *Lactobacillus janniae*, and *Lactococcus lactis* of 2:1:3; Temperature adjusted back to 37℃±0.5℃, dissolved oxygen increased to 1.5-2.0 mg / L, pH stabilized at 4.3-4.5; 0.1% whey protein concentrate was added to the culture medium.

[0192] Fifth stage (48-60h): Inoculation with *Lactobacillus gasseri*, *Lactobacillus casei*, *Lactobacillus dermalensis*, and *Enterococcus lactis*; Inoculation amount: 2% mother culture, with the weight ratio of *Lactobacillus gasseri*, *Lactobacillus casei*, *Lactobacillus dermalensis*, and *Enterococcus lactis* being 5:3:2:5; Temperature reduced to 30℃±1℃, dissolved oxygen naturally stabilized, and pH naturally stabilized; The bacterial culture was microencapsulated according to the method of Preparation Example 1, and 0.02% phytase was added to the culture medium;

[0193] 4) Sterilization: Flash sterilization at 85℃ for 15s; filtration, followed by stabilization treatment at 4℃ for 24h to obtain lactobacillus / soy milk fermentation product filtrate.

[0194] Preparation Examples 8-13

[0195] A selective estrogen receptor modulator, the preparation method of which is as follows:

[0196] According to the proportions in Table 1, the raw materials are mixed to obtain a selective estrogen receptor modulator.

[0197] Table 1. Raw material ratio table for preparation examples 8-13 (100g)

[0198]

[0199] Among them, propylene glycol / kudzu root extract, dodder seed extract, chicken blood vine extract, flaxseed oil / yam root extract, oregano leaf extract, and lactic acid bacteria fermentation product were derived from preparation examples 2-7, respectively.

[0200] Example

[0201] Examples 1-3

[0202] An ovarian maintenance and antibacterial gel, the preparation method of which is as follows:

[0203] S1. According to the raw material ratio in Table 2, mix the materials of phase A, and heat to 90°C while stirring. After stirring until all materials are completely melted, start cooling.

[0204] S2. When the temperature drops to 45℃, add phases B and C and stir for 30 minutes;

[0205] S3. Stop stirring, take samples for testing, and filter out the material after all physical and chemical indicators have passed the test.

[0206] Table 2 Raw material ratio table for Examples 1-3 (100g)

[0207]

[0208] The selective estrogen receptor modulator was derived from Preparation Example 8, and the extract of *Solanum nigrum* was derived from Preparation Example 1.

[0209] Examples 4-8

[0210] Unlike Example 2, the selective estrogen receptor modulators in Examples 4-8 were derived from Preparation Examples 9-13, respectively.

[0211] Comparative Example

[0212] Comparative Example 1

[0213] Unlike Example 2, the raw material ratios in Comparative Example 1 are shown in Table 2.

[0214] Comparative Example 2

[0215] Unlike Example 2, the raw material ratios in Comparative Example 2 are shown in Table 2.

[0216] Comparative Example 3

[0217] Unlike Example 2, in Comparative Example 3, an equal amount of selective estrogen receptor modulator was used to replace the extract of *Solanum nigrum*.

[0218] Comparative Example 4

[0219] Unlike Example 2, in Comparative Example 4, an equal amount of *Solanum nigrum* extract was used to replace the selective estrogen receptor modulator.

[0220] Application examples

[0221] Application Examples 1-8

[0222] The patient applied the gel obtained in Examples 1-8 into the vagina.

[0223] Comparative Application Examples

[0224] Compare and contrast examples 1-4

[0225] Patients applied the gels obtained in proportions 1-4 into the vagina.

[0226] Performance testing

[0227] I. Antibacterial testing was performed on the antibacterial gels obtained in the examples and comparative examples:

[0228] 1. Activation of bacterial strains: Escherichia coli, Staphylococcus aureus, and Lactobacillus stored at 4°C were transferred to fresh nutrient agar slants and incubated at 37°C for 24 hours.

[0229] 2. Preparation of bacterial suspension: The bacterial growth on the slant was washed twice with 10 mL of sterile physiological saline and transferred to an Erlenmeyer flask containing glass beads. The flask was placed on a shaker and shaken at 200 r / min for 10 min. The concentration of the bacterial suspension was adjusted to 106 cfu / ml.

[0230] 3. Preparation of antibacterial plates: Dispense 20 ml of melted nutrient agar medium into each test tube, add rubber stoppers, bundle 7 tubes together, sterilize at 121℃ for 20 min, and after sterilization and cooling, place in a 50℃ constant temperature water bath for incubation. On a clean bench, add 1 mL of bacterial suspension to each sterilized petri dish, pour in 20 mL of the suspension, mix thoroughly, blow off any condensate, and allow to solidify.

[0231] 4. Oxford Cup Method: Using sterile forceps, remove a sterile Oxford cup, briefly blanch it over an alcohol lamp flame, then place it vertically on the culture medium surface and gently press to ensure there are no gaps between the bottom of the cup and the medium. Place five Oxford cups on each plate, injecting 200 μL of gel into each cup, with physiological saline as a control in between. The gel must not overflow. Perform three replicates for each gel type. Incubate at 37°C for 24 hours, observe and record the diameter of the inhibition zone. The test results are shown in Table 3.

[0232] II. 120 premenopausal women aged 45 and above with symptoms of premature ovarian failure were randomly divided into 12 groups of 10 each. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were measured. Treatment was then administered according to the methods described in the application case and the control case. FSH and LH levels were then measured again. The method was as follows: blood was collected on days 2-4 of the menstrual cycle (early follicular phase), and serum FSH and LH concentrations were measured using chemiluminescence immunoassay or radioimmunoassay. The growth rate of FSH and LH concentrations was calculated as follows: growth rate = (post-treatment concentration - pre-treatment concentration) / pre-treatment concentration * 100%. The results are shown in Table 4.

[0233] Table 3 Results of Antibacterial Efficacy Test

[0234]

[0235] Based on Examples 1-8 and Comparative Examples 1-4, and in conjunction with Table 34, it can be seen that the gels in Examples 1-8 have better antibacterial effects against Escherichia coli and Staphylococcus aureus than those in Comparative Examples 1-4. This indicates that the antibacterial gel obtained in this application can effectively inhibit harmful bacteria. When applied to the vagina, it can effectively kill invading harmful bacteria and inhibit the occurrence of inflammation.

[0236] Table 4 Results of Ovarian Maintenance Performance Test

[0237]

[0238] Combining application examples 1-8 with comparative application examples 1-4, and referring to Table 4, it can be seen that the growth rates of FSH and LH in application examples 1-8 are higher than those in comparative application examples 1-4. This indicates that when the gel obtained in this application is applied to premature ovarian failure, it has a stronger effect on increasing FSH and LH, effectively improving premature ovarian failure. This may be because the gel obtained within the formulation range of this application has a stronger effect on improving premature ovarian failure.

[0239] Combining Application Examples 2 and 4-8, and referring to Table 4, it can be seen that the gel obtained in Application Example 6 has a stronger effect on increasing FSH and LH when applied to prematurely aging ovaries. This may be because the selective estrogen receptor modulator plays a role in ovarian maintenance and antibacterial scenarios through multiple pathways such as phytoestrogenic mimicry, hormone metabolism regulation, anti-oxidative stress, microecological regulation, and direct pathogen inhibition. The selective estrogen receptor modulator obtained in the formulation of Application Example 6 has a better effect.

[0240] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. An ovary maintenance bacteriostatic gel, characterized by, The A component, the B component and the C component are included; The A component includes the following raw materials by weight: deionized water 64-84 parts, preservative synergist EDTA-2Na 0.05-0.15 parts, Carbopol U-30 0.6-0.9 parts, super high molecular weight hyaluronic acid with a molecular weight of 2.2 million DA 0.06-0.12 parts, hydrogel 3-7 parts, glycerol 3-6, propylene glycol 3-6; The B component includes the following raw materials by weight: selective estrogen receptor modulator 3-7 parts, extract of Sphaeranthus indicus 0.5-2.5 parts; The C component includes the following raw materials by weight: silver ion solution 0.1-0.3 parts, preservative 0.3-0.6 parts, Tween-20 0.05-0.15 parts, triethanolamine 0.05-0.25 parts; The selective estrogen receptor modulator is composed of the following raw materials by weight: propylene glycol / Pueraria lobata extract 1-3 parts, Cuscuta chinensis seed extract 0.5-2 parts, Spatholobus suberectus extract 0.5-1.5 parts, linseed oil / Dioscorea opposita root extract 1-2.5 parts, tocopheryl acetate 0.1-0.5 parts, thiamine hydrochloride 0.05-0.2 parts, pyridoxine 0.05-0.2 parts, Origanum vulgare leaf extract 0.3-1 parts, lactic acid bacteria fermentation product 2-5 parts.

2. An ovary maintenance bacteriostatic gel according to claim 1, wherein The weight ratio of the propylene glycol / Pueraria lobata extract, Cuscuta chinensis seed extract, Spatholobus suberectus extract, and linseed oil / Dioscorea opposita root extract is 3:1.2:1:2.

5.

3. The ovary maintenance bacteriostatic gel according to claim 1, wherein The preparation method of the extract of Sphaeranthus indicus is as follows: 1) Raw material preparation Sphaeranthus indicus is dried and crushed; 2) First extraction The crushed Sphaeranthus indicus is soaked for 30-35 min, distilled at a pressure of 0.1-0.2 MPa and a temperature of 96-100℃, and the 3-hour distillate is collected, and the residue is collected; the distillate is subjected to oil-water separation, and the volatile oil in the upper layer is collected and stored in the dark; 3) Second extraction The residue after distillation in step 2) is dehydrated by pressing, ethanol is added, and ultrasonic treatment is performed at 40-45℃, followed by reflux extraction at 80-83℃ for 2 hours each time; after reflux extraction, the filtrate is combined while hot, and the filter residue is collected; 4) Third extraction The filter residue obtained in step 3) is dried by evaporating ethanol, water is added, and stirring and leaching are performed at 60-65℃ for 4 hours, and the supernatant is obtained by centrifugation; 5) Emulsification The filtrate obtained in step 3) and the supernatant obtained in step 4) are combined, and the volatile oil obtained in step 1) and an emulsifying agent are added, and homogenized to a particle size of ≤5 μm; 6) Filtration, concentration and drying to obtain the extract of Sphaeranthus indicus.

4. An ovary maintenance bacteriostatic gel according to claim 3, wherein The weight ratio of the emulsifying agent to the volatile oil in step 5) is 1:

1.

5. The ovary maintenance bacteriostatic gel according to claim 1, wherein The hydrogel is Polymatrix 7 hydrogel.

6. A method of preparing the ovary maintenance bacteriostatic gel as claimed in any one of claims 1 to 5, characterized in that, The following steps are included: S1. Mix the A phase materials, stir while warming to 85-95℃, and start cooling after the materials are completely dissolved; S2. When the temperature drops to 40-50℃, add the B and C phase materials, and stir for 30 minutes; S3. Stop stirring, take the sample for testing, and filter the sample after the physicochemical indicators are qualified.

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