A hydrogen bond self-assembled alginate polysaccharide gynecological gel and a preparation method thereof

Abstract

The present application belongs to the technical field of gynecological gel, and particularly relates to a fucoidan-based gynecological gel based on hydrogen bond self-assembly and a preparation method thereof. The fucoidan-based gynecological gel is composed of the following components: carbomer, modified fucoidan, PDRN, a pH stabilizer and a protective agent; the modified fucoidan is a fucoidan gel grafted with carboxymethyl chitosan. The present application introduces the chemically modified fucoidan and PDRN into the carbomer network through hydrogen bond self-assembly technology, utilizes the complementary hydrogen bonds of the three in different scales, realizes spontaneous and ordered intermolecular assembly, constructs a composite gel with a multiple cross-linking interpenetrating structure, and is supplemented with a system protective agent. Not only is the function of slow release and synergy realized, but also the structural stability and film-forming compactness of the gel in an acidic environment are significantly improved, thereby providing a new solution for the auxiliary treatment of chronic cervicitis and vaginitis.

Description

Technical Field

[0001] This invention belongs to the field of gynecological gel technology, specifically relating to a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides and its preparation method. Background Technology

[0002] Chronic cervicitis and vaginitis are common diseases of the female reproductive system. Pathogens include chlamydia, mycoplasma, bacteria, fungi, and trichomonas, often manifesting as increased vaginal discharge, vulvar itching and burning, and lower abdominal distension, severely impacting patients' quality of life. Currently, most gynecological dressings on the market only focus on single antibacterial or anti-inflammatory effects, lacking comprehensive regulation of the vaginal microenvironment, such as pH, moisture, and film integrity. They also generally suffer from drawbacks such as rapid release of active ingredients, short-lived local action, and insufficient synergistic wound repair capabilities.

[0003] Marine-derived brown algae polysaccharides possess natural antibacterial and moisturizing properties as well as excellent biocompatibility, while polydeoxyribonucleotides (PDRN) have anti-inflammatory and tissue repair functions that promote cell regeneration. Both have application potential in the field of gynecological mucosal care. However, when these two active components are combined into the same gel formulation, the limited chemical activity of brown algae polysaccharides makes it difficult to efficiently couple with other functional components. Carbomer matrix gels are prone to structural collapse under the acidic conditions of the vagina, resulting in uneven film formation and pH fluctuations, directly affecting the actual therapeutic effect. Existing carbomer gynecological gels often involve the direct physical blending of brown algae polysaccharides and carboxymethyl chitosan. The weak interaction between polysaccharide molecules leads to poor gel network stability, making them prone to water separation and rheological property degradation during storage. Some gels rely on chemical cross-linking agents to construct the network, and the residue of these cross-linking agents can pose a risk of vaginal mucosal irritation. In addition, existing hydrogen-bonded self-assembled supramolecular hydrogels are mostly used in skin wound dressings. Most hydrogen bond networks cannot withstand high-temperature and humid heat terminal sterilization, and the gel structure is easily destroyed after sterilization, making it difficult to meet the sterility requirements of gynecological preparations. PDRN is mostly added to existing gels through simple physical blending and has not been effectively combined with the polysaccharide network. The active substances are released too quickly, and the duration of action is limited. Summary of the Invention

[0004] The purpose of this invention is to provide a gynecological gel based on hydrogen-bonded self-assembly of brown algae polysaccharides, its preparation method, and its applications, thereby overcoming the shortcomings of existing technologies. Through hydrogen-bonded self-assembly technology, chemically modified brown algae polysaccharides and PDRN are jointly introduced into a carbomer network. Utilizing complementary hydrogen bonds at different scales (hydroxyl / carboxyl groups of brown algae polysaccharides, carboxyl groups of carbomer, and phosphate groups and bases of PDRN), spontaneous and ordered intermolecular assembly is achieved, constructing a composite gel with a multi-layered cross-linked interpenetrating structure, supplemented by a system protectant. This not only achieves synergistic sustained-release function but also significantly improves the structural stability and film density of the gel in acidic environments, providing a novel solution for the adjunctive treatment of chronic cervicitis and vaginitis.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows: In a first aspect, the present invention provides a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides, the gynecological gel being composed of the following components: Carbomer, modified brown algae polysaccharide, polydeoxyribonucleic acid PDRN, pH stabilizer and protectant; The mass ratio of carbomer, modified brown algae polysaccharide, and polydeoxyribonucleotide PDRN is 1:(0.1~2):(0.01~2). For example, the mass ratio of carbomer, modified brown algae polysaccharide, and polydeoxyribonucleic acid PDRN is any one of the following values ​​or ranges: 1:0.2:0.2, 1:0.5:0.5, 1:1:1, or 1:2:2.

[0006] The modified brown algae polysaccharide is a brown algae polysaccharide gel grafted with carboxymethyl chitosan.

[0007] In the formulation of the brown algae polysaccharide gynecological gel, carbomer serves as the matrix component, providing the basic rheological properties of the gel and ensuring its adhesion and retention in the vaginal area. Carboxymethyl chitosan-grafted modified brown algae polysaccharides contain numerous hydrogen bonding sites, which can interact with carbomer through hydrogen bonds to jointly construct a stable three-dimensional interpenetrating network, eliminating the need for additional chemical cross-linking agents and avoiding the risk of mucosal irritation caused by cross-linking agent residues. Polydeoxyribonucleotides (PDRN) are encapsulated within the interpenetrating network structure, enabling slow release and continuous vaginal mucosal repair. A pH stabilizer maintains the system in a slightly acidic vaginal environment, preserving the original vaginal microecological balance. A protectant further enhances the gel network's resistance to sterilization, ensuring that the gel structure is not damaged and the active substances are not degraded after sterilization. All components work synergistically to give this gynecological gel excellent adhesion, storage stability, and mucosal repair effects.

[0008] Secondly, the present invention provides the preparation of a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides, comprising the following steps: Carbomer solution was added to the heated modified brown algae polysaccharide and stirred until homogeneous. Then, polydeoxyribonucleic acid (PDRN) solution was added and stirred until homogeneous. The mixture was cooled to room temperature and stirred again to obtain a three-dimensional interpenetrating network gel. A pH stabilizer was added to the obtained three-dimensional interpenetrating network gel to adjust the pH of the system to acidic. A protective agent was then added and mixed evenly. The mixture was then filled and sterilized to obtain the brown algae polysaccharide gynecological gel.

[0009] This preparation method relies on hydrogen bond self-assembly to construct a three-dimensional interpenetrating network gel, without the need for external chemical cross-linking agents, thus avoiding the irritation of vaginal mucosa caused by cross-linking agent residues. The process conditions are mild, which is conducive to protecting the active ingredient PDRN. By adjusting the pH to acidic, it matches the slightly acidic physiological environment of the vagina. With the addition of a protectant and terminal sterilization treatment, the gel network structure remains stable after sterilization, and it is not prone to collapse or water separation. The finished product meets the sterility requirements of gynecological preparations.

[0010] Thirdly, this invention provides the application of hydrogen bond-based self-assembled brown algae polysaccharide gynecological gel in the preparation of adjunctive treatment products for chronic cervicitis and vaginitis.

[0011] This invention utilizes the cell regeneration / anti-inflammatory activity of PDRN and the strong moisturizing / broad-spectrum antibacterial properties of fucoidan to achieve gradient sustained release of "dual active ingredients" through a hydrogen bond network - the outer layer rapidly releases fucoidan for rapid antibacterial action, while the inner layer continuously releases PDRN for long-term repair, significantly improving the healing efficiency of chronic cervicitis wounds.

[0012] The beneficial effects of this invention are: (1) The present invention uses hydrogen bond self-assembly to form a gel, without the need for external chemical crosslinking agents, thus avoiding the irritation of vaginal mucosa caused by crosslinking agent residue; the overall preparation process is mild, which can protect PDRN and polysaccharide active substances from being destroyed. With the help of the protective agent, the finished product can still maintain the gel network structure and the efficacy of active ingredients after high temperature sterilization. The process is simple to operate, with good batch reproducibility, and achieves a comprehensive improvement in the formulation's high efficiency, stability and physiological environment adaptability, which is convenient for large-scale production and application.

[0013] (2) The pH of the gel system of this invention is stably maintained at 3.8-5.5, which is compatible with the slightly acidic environment of the vagina and will not disturb the microecological balance of beneficial bacteria such as lactobacilli in the vagina. The three-dimensional interpenetrating network formed by hydrogen bond self-assembly gives the gel excellent film-forming properties and mucosal adhesion ability. It can form a uniform protective film on the vaginal wall and cervix, and can adsorb wound exudate and pathogen metabolites, reducing local irritation. The synergistic effect of the brown algae polysaccharide and PDRN component can significantly inhibit the release of inflammatory factors such as TNF-α and IL-6 induced by lipopolysaccharide. The wound healing effect is better than that of a single component. It is not a simple superposition of the effects of each component, avoiding the risk of dilution and antagonism between components, solving the formulation compatibility problem between active substances and excipients, and integrating multiple effects of anti-inflammatory and mucosal repair. It is suitable for the adjunctive intervention of vaginitis and chronic cervicitis. Attached Figure Description

[0014] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0015] Figure 1 In vitro release performance diagram of PDRN in Example 1; Figure 2 The effects of different samples diluted 100-fold on cellular inflammatory factors were investigated. In this study, A represents the effect on cellular inflammatory factor IL-6, and B represents the effect on cellular inflammatory factor TNF-α. Figure 3 The effect of diluting different samples by 100-fold on cell healing rate; in, Figure 2 and Figure 3 In the composition, composition 1 consists of carbomer and PDRN, and composition 2 consists of carbomer, PDRN, and fucoidan; compared with the control group, P ### <0.001, compared with the model group, P <0.01, P <0.001). Detailed Implementation

[0016] Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be construed as limiting the scope of the invention. Specific conditions not specified in the embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Components whose manufacturers are not specified are all commercially available conventional products.

[0017] In a first aspect, the present invention provides a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides, which is composed of the following components: carbomer, modified brown algae polysaccharides, polydeoxyribonucleotides (PDRN), pH stabilizers, and protectants.

[0018] The mass ratio of carbomer, modified brown algae polysaccharide, and polydeoxyribonucleotide PDRN is 1:(0.1~2):(0.01~2).

[0019] The modified brown algae polysaccharide is a brown algae polysaccharide grafted with carboxymethyl chitosan.

[0020] This invention is the first to use carboxymethyl chitosan to amidate graft modification of brown algal oligosaccharides, which not only improves the water solubility and reactivity of brown algal polysaccharides, but also introduces abundant amide bonds and amino / carboxyl pairs, providing multiple interaction sites for subsequent hydrogen bond self-assembly.

[0021] In some other embodiments, the modified brown algae polysaccharide is prepared by adding a carboxymethyl chitosan aqueous solution to an aqueous solution of brown algae polysaccharide, and then adding a condensing agent and triethylamine in sequence, stirring and reacting at room temperature to obtain the product.

[0022] This modified brown algae polysaccharide was prepared using a room-temperature liquid-phase amide grafting reaction. The reaction conditions are mild, requiring no high temperature or pressure, thus avoiding thermal degradation of the polysaccharide molecules. Through the combined action of a condensing agent and triethylamine, amidation covalent grafting of brown algae polysaccharide and carboxymethyl chitosan was achieved, introducing numerous hydrogen bonding sites onto the molecular chain. This lays the molecular foundation for subsequent hydrogen-bonded self-assembly with carbomer to construct a three-dimensional interpenetrating gel network. The overall process is simple to operate, requiring no complex post-processing. The grafted polysaccharide retains the bioactivity of both brown algae polysaccharide and carboxymethyl chitosan, exhibits good biocompatibility, and introduces no toxic cross-linking components, making it suitable for use in gynecological mucosal preparations.

[0023] In some other embodiments, the mass ratio of the brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:(0.01~2):(0.01~2). Preferably, the mass ratio of brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:1:(0.2~1). The molecular weight of the brown algae polysaccharide is <6000, and the molecular weight of the carboxymethyl chitosan is 2000~100000; The condensing agent is selected from 2-chloro-1-methylpyridinium iodide (CMPI) or 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium hydrochloride (DMTMM).

[0024] In some other embodiments, the pH stabilizer is selected from one or more of NaOH, KOH, triethanolamine, and triethylamine.

[0025] The pH stabilizer is added at a rate of 0.1 wt% to 1.0 wt% of the total mass of the brown algae polysaccharide gynecological gel.

[0026] In some other embodiments, the protective agent is a PBS solution with a pH of 7.0 to 7.5.

[0027] The amount of the protective agent added is 0.05wt% to 10wt% of the total mass of the brown algae polysaccharide gynecological gel.

[0028] Secondly, the present invention provides the preparation of a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides, comprising the following steps: Carbomer solution was added to the heated modified brown algae polysaccharide and stirred until homogeneous. Then, polydeoxyribonucleic acid (PDRN) solution was added and stirred until homogeneous. The mixture was cooled to room temperature and stirred again to obtain a three-dimensional interpenetrating network gel. A pH stabilizer was added to the obtained three-dimensional interpenetrating network gel to adjust the pH of the system to acidic. A protective agent was then added and mixed evenly. After filling and sterilization, the brown algae polysaccharide gynecological gel was obtained.

[0029] This invention proposes a ternary hydrogen-bonded interpenetrating network system of "modified brown algae polysaccharide-carbomer-PDRN". By utilizing the complementary hydrogen bonds of the three at different scales (hydroxyl / carboxyl groups of brown algae polysaccharide, carbomer carboxyl groups, phosphate groups and bases of PDRN), spontaneous and ordered intermolecular assembly is achieved to form a dense and flexible gel framework, which is significantly superior to traditional simple physical mixing.

[0030] In some other embodiments, the heating temperature of the modified brown algae polysaccharide is 40~80°C, and the carbomer solution is prepared by low-temperature swelling or heating and dissolving of carbomer.

[0031] In some other embodiments, the concentration of the carbomer solution is 0.1 wt% to 1.0 wt%, the concentration of the polydeoxyribonucleic acid (PDRN) solution is 0.01 wt% to 0.8 wt%, the concentration of the pH stabilizer is 0.1 wt% to 1.0 wt%, and the concentration of the protectant is 0.05 wt% to 10 wt%.

[0032] For example, modified fucoidan is added to a carbomer solution at 40-80°C and stirred at a constant temperature for 1-2 hours; then PDRN (0.01%-0.8% by mass) is added under constant temperature conditions, and stirring continues for 1-6 hours; after natural cooling to room temperature, it is stirred for another 0.5-1 hour to form a uniform, transparent three-dimensional interpenetrating network gel. The mass ratio of carbomer, modified fucoidan, and PDRN is 1:(0.1-2):(0.01-2). A pH stabilizer (one or more of NaOH, KOH, or triethanolamine and triethylamine, at a mass concentration of 0.1%-1.0%) is added to the three-dimensional interpenetrating network gel to adjust the pH to 3.8-5.5, followed by the addition of a protective agent (PBS solution with pH=7.4, at a concentration of 0.05%-10%). After mixing, the mixture is filled into containers and sterilized at 121°C for 20 minutes using a moist heat method to obtain the finished gynecological gel.

[0033] In some other embodiments, the pH of the system is adjusted to 3.8-5.5, and the sterilization treatment is performed at 121°C for 10-30 minutes. The resulting gel has a stable pH value of 3.8-5.5, which matches the normal physiological environment of the vagina and does not disrupt the ecology of beneficial bacteria such as lactobacilli. The sterilization treatment can effectively kill microorganisms in the gel system, ensuring that the finished product meets the sterility requirements of vaginal mucosal drug delivery formulations.

[0034] Thirdly, this invention provides the application of hydrogen-bonded self-assembled brown algae polysaccharide gynecological gel in the preparation of adjunctive treatment products for chronic cervicitis and vaginitis. The product retains its gel morphology and activity after high-temperature sterilization. The process is simple and reproducible, providing an innovative dosage form with antibacterial, anti-inflammatory, and healing-promoting functions for the adjunctive treatment of chronic cervicitis and vaginitis.

[0035] Example 1 A gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides and its preparation method, comprising the following steps: (1) Synthesis of modified brown algae polysaccharide (amidation grafting): Prepare an aqueous solution of brown algae polysaccharide (molecular weight <6000) with a mass concentration of 5%, stir for 1 hour, slowly add carboxymethyl chitosan aqueous solution (mass concentration 0.5%, molecular weight 10000), and add condensing agent (2-chloro-1-methylpyridinium iodide CMPI) and appropriate amount of triethylamine in sequence. Stir and react at room temperature for 1 hour to obtain carboxymethyl chitosan grafted modified brown algae polysaccharide gel intermediate.

[0036] The mass ratio of brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:1:0.2.

[0037] (2) Dissolution of carbomer matrix: Carbomer was added to purified water and dissolved overnight at low temperature or at 80°C to obtain a carbomer solution with a mass concentration of 0.3%.

[0038] (3) Preparation of hydrogen bond self-assembled composite gel: The modified brown algae polysaccharide obtained in step (1) was added to the carbomer solution in step (2) at 80°C and stirred at a constant temperature for 1 hour; then polydeoxyribonucleotide (PDRN, mass concentration 0.3%) was added under constant temperature conditions and stirred for another 2 hours; it was naturally cooled to room temperature and stirred for another 0.5 hours to form a uniform and transparent three-dimensional interpenetrating network gel.

[0039] The mass ratio of carbomer, modified brown algae polysaccharide, and PDRN is 1:0.2:0.2.

[0040] (4) pH adjustment: Add the pH stabilizer triethanolamine (0.2% by mass) to the three-dimensional interpenetrating network gel in step (3) to adjust the pH of the system to 4.96 to adapt to the slightly acidic environment of a healthy vagina.

[0041] (5) Sterilization and packaging: Add a protective agent (PBS solution with pH=7.4, content 0.1%) to the system in step (4), mix well and fill into the container, sterilize at 121°C for 20 minutes to obtain the finished gynecological gel (labeled as composition 2).

[0042] Example 2 A gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides and its preparation method, comprising the following steps: (1) Synthesis of modified brown algae polysaccharide (amidation grafting): Prepare an aqueous solution of brown algae polysaccharide (molecular weight <6000) with a mass concentration of 10% and stir for 2 hours. Slowly add an aqueous solution of carboxymethyl chitosan (mass concentration 1%, molecular weight 20000) and add a condensing agent (4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium hydrochloride DMTMM) and an appropriate amount of triethylamine in sequence. Stir the reaction at room temperature for 3 hours to obtain a carboxymethyl chitosan grafted modified brown algae polysaccharide gel intermediate.

[0043] The mass ratio of brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:1:0.5.

[0044] (2) Dissolution of carbomer matrix: Carbomer was added to purified water and dissolved overnight at low temperature or at 60°C to obtain a carbomer solution with a mass concentration of 0.5%.

[0045] (3) Preparation of hydrogen bond self-assembled composite gel: The modified brown algae polysaccharide obtained in step (1) was added to the carbomer solution in step (2) at 60°C and stirred at a constant temperature for 1.5 hours; then polydeoxyribonucleotide (PDRN, mass concentration 0.5%) was added under constant temperature conditions and stirred for 4 hours; it was naturally cooled to room temperature and stirred for another 0.5 hours to form a uniform and transparent three-dimensional interpenetrating network gel.

[0046] The mass ratio of carbomer, modified brown algae polysaccharide, and PDRN is 1:0.5:0.5.

[0047] (4) pH adjustment: Add pH stabilizer KOH at a mass concentration of 0.1% to the three-dimensional interpenetrating network gel in step (3) to adjust the pH of the system to 5.00 to adapt to the slightly acidic environment of a healthy vagina.

[0048] (5) Sterilization and packaging: Add a protective agent (PBS solution with pH=7.4, content 0.2%) to the system in step (4), mix well and fill into the container, sterilize at 121°C for 20 minutes to obtain the finished gynecological gel.

[0049] Example 3 A gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides and its preparation method, comprising the following steps: (1) Synthesis of modified brown algae polysaccharide (amidation grafting): Prepare an aqueous solution of brown algae polysaccharide (molecular weight <6000) with a mass concentration of 20% and stir for 3 hours. Slowly add an aqueous solution of carboxymethyl chitosan (mass concentration 2%, molecular weight 100000) and add a condensing agent (4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholinium hydrochloride DMTMM) and an appropriate amount of triethylamine in sequence. Stir the reaction at room temperature for 1 to 5 hours to obtain a carboxymethyl chitosan grafted modified brown algae polysaccharide gel intermediate.

[0050] The mass ratio of brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:1:1.

[0051] (2) Dissolution of carbomer matrix: Carbomer was added to purified water and dissolved overnight at low temperature or at 40°C to obtain a carbomer solution with a mass concentration of 0.8%.

[0052] (3) Preparation of hydrogen bond self-assembled composite gel: The modified brown algae polysaccharide obtained in step (1) was added to the carbomer solution in step (2) at 40°C and stirred at a constant temperature for 2 hours; then polydeoxyribonucleotide (PDRN, mass concentration 0.8%) was added under constant temperature conditions and stirred for 6 hours; it was naturally cooled to room temperature and stirred for another hour to form a uniform and transparent three-dimensional interpenetrating network gel.

[0053] The mass ratio of carbomer, modified brown algae polysaccharide, and PDRN is 1:1:1.

[0054] (4) pH adjustment: Add pH stabilizer NaOH (0.1% by mass) to the three-dimensional interpenetrating network gel in step (3) to adjust the pH of the system to 5.01 to adapt to the slightly acidic environment of a healthy vagina.

[0055] (5) Sterilization and packaging: Add a protective agent (PBS solution with pH=7.4, content 1%) to the system in step (4), mix well and fill into the container, sterilize at 121°C for 20 minutes to obtain the finished gynecological gel.

[0056] Comparative Example 1 Unlike Example 1, step (1) was omitted, and the carbomer solution was directly stirred and mixed with PDRN to carry out the reaction. Other steps remained unchanged, and the product was obtained and labeled as Composition 1 (the components are carbomer and PDRN).

[0057] Specifically, the preparation method of composition 1 is as follows: weigh an appropriate amount of carbomer, add an appropriate amount of purified water, stir until dispersed at room temperature, then add to a 65°C water bath for 1 hour, let it stand at room temperature, weigh an appropriate amount of PDRN, add it to the carbomer gel, adjust the pH to 5.05 with triethanolamine, add PBS system protectant, and sterilize at 121°C for 20 minutes.

[0058] Comparative Example 2 Unlike Example 1, carbomer, fucoidan, and PDRN were used as dressings, respectively.

[0059] ①Preparation of carbomer: Weigh an appropriate amount of carbomer, add purified water, stir until dispersed at room temperature, then add to a 65℃ water bath for 1 hour, let it stand at room temperature, adjust the pH to 5.01 with triethanolamine, and sterilize at 121℃ for 20 minutes.

[0060] ②Preparation of PDRN: Take an appropriate amount of PDRN, add PBS and stir until dissolved, then sterilize at 121℃ for 20 min.

[0061] ③ Preparation of modified brown algae polysaccharide: Take an appropriate amount of modified brown algae polysaccharide gel and sterilize it at 121℃ for 20 min.

[0062] Comparative Example 3 Unlike Example 1, steps (1)-(3) are omitted, and a one-step method is used to directly physical blend brown algae polysaccharide, carboxymethyl chitosan and carbomer, while other steps remain unchanged.

[0063] Performance testing The samples prepared in the examples and comparative examples were subjected to performance tests, and the test results are shown in Tables 1 and 2. Viscosity was determined according to the rotational viscometer method in General Chapter 0633 of the Pharmacopoeia of the People's Republic of China (2025 Edition, Part IV).

[0064] The pH value was determined according to the pH value determination method in General Chapter 0631 of the Pharmacopoeia of the People's Republic of China (2025 Edition, Part IV).

[0065] The gel strength was determined according to the method specified in General Chapter 0634 of the Pharmacopoeia of the People's Republic of China (2025 Edition, Part IV).

[0066] The structural stability under acidic conditions was characterized by rheological properties, in accordance with the method in General Technical Requirements and Guidelines 0633 of the Pharmacopoeia of the People's Republic of China (2025 Edition, Part IV) for viscosity determination (Rheometer Method III).

[0067] Centrifugation stability: Centrifuge at 3000 rpm for 30 minutes and observe whether there is stratification or precipitation.

[0068] Test method for film density and film stability: Take an appropriate amount of gel, place it on a horizontal glass slide, and spread it into a thin layer of gel about 2 mm thick. The area of ​​the thin layer is equivalent to the area of ​​the coverslip. The glass slide is held upright at a 90° angle to the horizontal plane for 1 minute. No shrinkage pores were observed in the coated gel layer, and no obvious flow phenomenon was observed in the gel layer.

[0069] Stability test: The gels obtained in Examples 1-3 and 3 comparative sample gels were placed at (25±0.1)℃ for 7 days and the above properties were tested in the same way. The results are shown in Table 1 and Table 2.

[0070] Table 1. Results of viscosity, gel strength, pH, and film density tests.

[0071] As shown in Table 1, the gels prepared in Examples 1-3 exhibited better viscosity, gel strength, and film density than the comparative examples, and their pH values ​​were also more stable. After being placed at (25 ± 0.1) °C for 7 days, the gel properties of the gels prepared in Examples 1-3 remained essentially unchanged, while the properties of the samples in the comparative examples all decreased to varying degrees, with significant pH value changes.

[0072] Table 2. Results of rheological properties and centrifugal stability tests

[0073] Table 2 shows that G' reflects the elasticity and solid-state characteristics of the gel. A higher G' indicates a denser cross-linked gel network and a more "rigid" structure. G'' reflects the viscosity and flow characteristics of the gel. The G' values ​​of Examples 1-3 are significantly greater than G'', generally more than three times higher. This means that a robust three-dimensional network structure has formed inside the gel, making it difficult to disrupt. The centrifugal stability test further confirms the stability of the gel structure prepared by this invention. After being placed at (25 ± 0.1) °C for 7 days, the properties of the gels prepared in Examples 1-3 remained essentially unchanged, while the rheological properties of the samples in the comparative examples decreased. The data in Tables 1 and 2 indicate that the internal structure of the samples in the comparative examples is unstable.

[0074] The PDRN in vitro release assay in Example 1 was performed according to GB / T 34796-2017 "Determination of Concentration and Purity of Nucleic Acids in Aqueous Solutions - Ultraviolet Spectrophotometry". The results are shown in [Figure Number]. Figure 1 .Depend on Figure 1 It was found that approximately 47% was released within 0.5 hours, exhibiting a burst release phenomenon. This initial burst release allows the PDRN concentration to quickly exceed the effective threshold, resulting in rapid onset of action. Approximately 85% was released after 1.5 hours, and about 90% after 4 hours: this indicates that PDRN is fully released within the expected usage time. The remaining approximately 10% may be tightly bound by the carbomer network or require a longer time to diffuse out. The study found that hydrogen-bonded self-assembled brown algae polysaccharide gynecological gel can stably maintain the in vitro release characteristics of PDRN.

[0075] 2. Cellular anti-inflammatory experiment Log-grown HaCaT cells were digested with trypsin and prepared into a cell suspension, with each well containing 1 × 10⁶ cells.5 Cells were seeded at a density of 2 mL of cell culture medium per well in 6-well plates and cultured for 24 h in a cell culture incubator. After 24 h, the cell culture medium was aspirated. 2 mL of fresh culture medium was added to the control group, while 2 mL of culture medium containing 1 μg / mL LPS was added to the model and experimental groups. Different concentrations of sample were added to the experimental groups. All cells were then cultured at 37°C for 24 h. After 24 h, the cell culture supernatant was collected from each group and centrifuged at 4000 r / min for 5 min. The cell culture supernatant was used for ELISA to detect the expression levels of IL-6 and TNF-α cytokines.

[0076] 3. Cell scratch assay (repair) Draw three lines across the back of a 6-well plate using a marker pen. Harvest HaCaT cells in logarithmic growth phase, digest with trypsin, centrifuge, and then... 6 Cells were seeded at a concentration of 100% in 6-well plates, with the seeding principle being that the confluence rate reached 100% overnight. After 24 hours, the 6-well plates were removed, and a ruler was placed perpendicular to the black line. Three scratches were gently made from left to right along the edge of the ruler using a 10 μL pipette tip. After scratching, different fields of view were selected for observation and photography under a microscope. After photography, low-serum culture medium was added to the control group. Low-serum culture medium containing 1 μg / mL EGF was added to the positive group, and low-serum culture medium containing different samples was added to the experimental groups. The cells were incubated at 37℃ in a 5% CO2 incubator for 24 hours. After 24 hours, the cells were removed, and the marked fields of view were observed and photographed under a microscope. The healing rate was calculated using the formula: Healing rate (%) = (Scratch area (0h) - Scratch area (24h)) / (Scratch area (0h)) × 100 Depend on Figure 2 It was found that the composition containing fucoidan, carbomer and PDRN (composition 2) had a significant synergistic inhibitory effect on the release of lipopolysaccharide-induced inflammatory factors (such as TNF-α and IL-6) (P<0.001), and was superior to composition 1 containing only carbomer and PDRN, which confirmed the combined synergistic effect of fucoidan and PDRN.

[0077] Depend on Figure 3 It is evident that Composition 2 exhibits superior healing-promoting activity compared to single components and conventional dressings, demonstrating that Composition 2 is not merely a simple additive effect of its components, but rather represents a substantial advancement. The synergistic effect of the components within the composition overcomes the risks of dilution or antagonism, successfully resolving the compatibility issues between the active ingredient and excipients, thus possessing beneficial pharmaceutical value.

[0078] In summary, this invention utilizes hydrogen-bonded self-assembly technology to introduce chemically modified fucoidan and PDRN into a carbomer network. By leveraging complementary hydrogen bonds at different scales (hydroxyl / carboxyl groups of fucoidan, carboxyl groups of carbomer, and phosphate and base groups of PDRN), spontaneous and ordered intermolecular assembly is achieved, constructing a composite gel with a multi-layered cross-linked interpenetrating structure, supplemented by a system protectant. This not only achieves synergistic sustained-release function but also significantly improves the gel's structural stability and film density under acidic conditions, providing a novel solution for the adjunctive treatment of chronic cervicitis and vaginitis.

[0079] The above are merely preferred embodiments of the present invention and are not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides, characterized in that, It consists of the following components: Carbomer, modified brown algae polysaccharide, polydeoxyribonucleic acid PDRN, pH stabilizer and protectant; The mass ratio of carbomer, modified brown algae polysaccharide, and polydeoxyribonucleotide PDRN is 1:(0.1~2):(0.01~2). The modified brown algae polysaccharide is a brown algae polysaccharide grafted with carboxymethyl chitosan.

2. The gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides as described in claim 1, characterized in that, The modified brown algae polysaccharide is prepared by adding carboxymethyl chitosan aqueous solution to brown algae polysaccharide aqueous solution, and then adding condensing agent and triethylamine in sequence, stirring and reacting at room temperature to obtain the product.

3. The gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides as described in claim 2, characterized in that, The mass ratio of the brown algae polysaccharide, condensing agent, and carboxymethyl chitosan is 1:(0.01~1):(0.01~1). The molecular weight of the brown algae polysaccharide is <6000, and the molecular weight of the carboxymethyl chitosan is 2000~100000; The condensing agent is selected from one or both of 2-chloro-1-methylpyridinium iodide (CMPI) or 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium hydrochloride (DMTMM).

4. The gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides as described in claim 1, characterized in that, The pH stabilizer is selected from one or more of NaOH, KOH, triethanolamine, and triethylamine; The pH stabilizer is added at a rate of 0.1 wt% to 1.0 wt% of the total mass of the brown algae polysaccharide gynecological gel.

5. The gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides as described in claim 1, characterized in that, The protective agent is a PBS solution with a pH of 7.0-7.5; The amount of the protective agent added is 0.05wt% to 10wt% of the total mass of the brown algae polysaccharide gynecological gel.

6. A method for preparing a gynecological gel based on hydrogen bond self-assembly of brown algae polysaccharides according to any one of claims 1-5, characterized in that, Includes the following steps: Carbomer solution was added to the heated modified brown algae polysaccharide and stirred until homogeneous. Then, polydeoxyribonucleic acid (PDRN) solution was added and stirred until homogeneous. The mixture was cooled to room temperature and stirred again to obtain a three-dimensional interpenetrating network gel. A pH stabilizer was added to the obtained three-dimensional interpenetrating network gel to adjust the pH of the system to acidic. A protective agent was then added and mixed evenly. The mixture was then filled and sterilized to obtain the brown algae polysaccharide gynecological gel.

7. The preparation method according to claim 6, characterized in that, The heating temperature of the modified brown algae polysaccharide is 40~80℃, and the carbomer solution is prepared by low-temperature swelling or heating and dissolving of carbomer.

8. The preparation method according to claim 6, characterized in that, The concentration of carbomer solution is 0.1 wt% to 1.0 wt%, the concentration of polydeoxyribonucleic acid (PDRN) solution is 0.01 wt% to 0.8 wt%, the concentration of pH stabilizer is 0.1 wt% to 1.0 wt%, and the concentration of protectant is 0.05 wt% to 10 wt%.

9. The preparation method according to claim 6, characterized in that, The pH of the system is adjusted to 3.8~5.5, and the sterilization process is performed at 121℃ for 10~30 min.

10. The use of the hydrogen bond-based self-assembled brown algae polysaccharide gynecological gel according to any one of claims 1-5 in the preparation of an adjunctive treatment product for chronic cervicitis and vaginitis.

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