A hard contact lens care solution and process for making same

Abstract

The present application relates to the technical field of contact lens disinfectant, and discloses a hard corneal contact lens care solution and a preparation process thereof. The hard corneal contact lens care solution comprises a multiple antibacterial system. The multiple antibacterial system comprises antibacterial peptides, beta-cyclodextrin-PHMB inclusion compound, zinc-doped mesoporous silica loaded with silver and HEMA-dopamine liposomes. In the zinc-doped mesoporous silica loaded with silver, mesoporous silica is used as a skeleton material, zinc is doped in the mesoporous silica skeleton material in the form of lattice doping, and silver is loaded in the mesoporous channel. The present application is prepared by using a multiple antibacterial system. Through the synergistic effect of the components of the prepared raw materials, the antibacterial performance is improved as a whole, the cytotoxicity is inhibited, good lubrication durability and active ingredient stability are achieved, and the stability and efficiency of the sterilization and disinfection effect are improved by combining a specific sterilization process.

Description

A rigid gas permeable contact lens care solution and its preparation process Technical Field

[0001] This invention relates to the field of contact lens disinfectant technology, and more specifically, to a rigid gas permeable contact lens care solution and its preparation process. Background Technology

[0002] Rigid Gas Permeable (RGP) contact lens solution is an ophthalmic medical preparation. It's a mixture of various components used to disinfect and sterilize rigid gas permeable (RGP) contact lenses. RGP lenses are contact lenses made of a rigid material, primarily used to correct high myopia, hyperopia, and other specific vision problems. Compared to soft contact lenses, they offer significantly better breathability and material stability, as well as excellent wetting and anti-deposition properties. RGP contact lens solution is commonly used for cleaning, disinfection, and storage of RGP lenses.

[0003] Existing rigid gas permeable contact lens solutions generally use a basic formula of poloxamer, HPMC, and disodium edetate mixed together, such as Weicon contact lens solution. Although these solutions have a more obvious cleaning effect and are widely used, they still have problems such as insufficient antibacterial and disinfection efficacy, low retention rate of effective ingredients, and sterilization process that is destructive to antibacterial effect.

[0004] Based on the above, a technical solution of introducing zinc into contact lens solutions is proposed to improve their antibacterial and other properties. For example, patent CN118078743A provides a mesoporous zinc oxide eye drop loaded with natamycin, its preparation method, and its application. The eye drop component includes mesoporous zinc oxide loaded with natamycin. Mesoporous zinc oxide can exert an antibacterial effect against fungal keratitis, but it may produce Zn... 2+ Sudden release can cause corneal cell toxicity, leading to a decrease in corneal cell survival.

[0005] In existing contact lens solutions containing zinc and other antibacterial ions, directly adding antibacterial ingredients can easily lead to increased eye irritation, inactivation of antibacterial ions, and increased cytotoxicity. Summary of the Invention

[0006] The purpose of this invention is to solve the problem that existing contact lens solutions that enhance antibacterial and disinfectant properties by introducing antibacterial ions also increase cytotoxicity. The invention proposes a solution that balances antibacterial enhancement and toxicity inhibition.

[0007] This invention is achieved through the following technical solution:

[0008] This invention provides a rigid gas permeable contact lens care solution, comprising a multi-antibacterial system;

[0009] The multiple antimicrobial system includes antimicrobial peptides, β-cyclodextrin-PHMB inclusion complex, zinc-doped mesoporous silica loaded with silver, and HEMA-dopamine liposomes;

[0010] In the zinc-doped mesoporous silica loaded with silver, mesoporous silica serves as the framework material, zinc is doped into the mesoporous silica framework material in the form of lattice doping, and silver is loaded into the mesoporous channels.

[0011] Preferably, the method for preparing the zinc-doped mesoporous silica loaded with silver is as follows:

[0012] Zinc-doped mesoporous silica was synthesized using the sol-gel method, then vacuum impregnated with silver nitrate solution; and finally calcined to obtain the zinc-doped mesoporous silica loaded with silver.

[0013] Preferably, in the method for preparing the zinc-doped mesoporous silica loaded with silver, the calcination temperature is 300-400℃ and the calcination time is 1-3h.

[0014] Preferably, in the zinc-doped mesoporous silica loaded with silver, the zinc content is 1.2-1.8% by mass and the silver loading is 0.1-0.2 wt%.

[0015] Preferably, in the zinc-doped mesoporous silicon dioxide loaded with silver, the mesopore diameter is 4-6 nm.

[0016] Preferably, in the β-cyclodextrin-PHMB inclusion complex, the molar ratio of β-cyclodextrin to PHMB is 1:2-4, and the mass ratio of PHMB to antimicrobial peptide is 0.4-1.2:1.

[0017] Preferably, in the HEMA-dopamine liposomes, the mass ratio of HEMA to dopamine is 2-4:1, and the particle size of the HEMA-dopamine liposomes is 80-120 nm.

[0018] The present invention also provides a preparation process for the above-mentioned rigid gas permeable contact lens care solution, comprising the following steps:

[0019] S1 Preliminary preparation of nursing solution:

[0020] Take raw materials containing multiple antibacterial systems, mix them well, and obtain the initial care solution;

[0021] S2 filtration process:

[0022] The initial nursing solution is filtered through a membrane.

[0023] S3 gradient sterilization process:

[0024] The following three-stage pulsed moist heat sterilization process is performed sequentially: 105℃ sterilization for 6-10 min, 121℃ sterilization for 1-3 min, and 115℃ sterilization for 4-6 min to obtain the rigid gas permeable contact lens care solution.

[0025] Preferably, in step S1, the raw materials, based on mass concentration, include:

[0026] 0.01-0.70% poloxamer;

[0027] 0.01-0.35% hydroxypropyl methylcellulose;

[0028] 0.01-0.10% disodium edetate;

[0029] 0.10-0.30% propylene glycol;

[0030] 0.05-0.30% buffer;

[0031] 0.0005-0.0020% antimicrobial peptides;

[0032] 0.0001-0.0020% β-cyclodextrin-PHMB inclusion complex;

[0033] 0.001-0.005% zinc-doped mesoporous silica loaded with silver;

[0034] 0.05-0.30% HEMA-dopamine liposomes;

[0035] The remainder consists of sodium hydroxide and solvent.

[0036] Preferably, in step S2, during membrane filtration, the temperature is controlled at 3-5℃ and the shear rate is 1200-1800 s. -1 .

[0037] The technical solution of the present invention has the following beneficial effects:

[0038] (1) In terms of composition, the present invention proposes to combine antimicrobial peptides, β-cyclodextrin-PHMB inclusion complex, and Zn-doped mesoporous SiO2 loaded with Ag. + It integrates a triple synergistic antibacterial system and incorporates HEMA-dopamine liposomes.

[0039] Specifically, antimicrobial peptides can significantly improve the efficiency of PHMB encapsulated by β-cyclodextrin entering bacterial cells through membrane perforation, while encapsulating PHMB with β-cyclodextrin can significantly reduce PHMB cytotoxicity; zinc doped into silver-loaded mesoporous silica particles can block bacterial resistance to Ag by inhibiting mexB gene expression. +The surface is coated with biomimetic sustained-release liposomes made of polydopamine, which have good adsorption properties and can achieve long-lasting lubrication.

[0040] This invention enhances antibacterial properties and inhibits cytotoxicity through the synergistic effect of multiple components, exhibiting good lubrication durability and stability of active ingredients. It effectively solves the current clinical problems of multidrug-resistant bacterial infections, biofilm formation on lens surfaces, and low retention rate of active ingredients in traditional care solutions.

[0041] (2) In terms of preparation process, dynamic gradient sterilization process is used to replace traditional filtration sterilization. Without affecting the degradation rate of PHMB and the encapsulation rate of liposomes, multi-stage gradient temperature sterilization process can greatly maintain the stability and integrity of the multi-antibacterial system, that is, solve the problems of low retention rate and poor stability that are common in traditional care solutions. The sterilization process used in this invention can enable the care solution to maintain long-term antibacterial performance. Detailed Implementation

[0042] Specific implementation details

[0043] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, they are performed according to conventional conditions or conditions recommended by the manufacturer; where the manufacturers of the instruments, equipment, reagents, or raw materials used are not specified, they are all conventional products that can be purchased commercially.

[0044] This invention provides a rigid gas permeable contact lens care solution, comprising the following components by mass concentration:

[0045] (1) 0.01-0.70% poloxamer;

[0046] (2) 0.01-0.35% hydroxypropyl methylcellulose (HPMC);

[0047] (3) 0.01-0.10% disodium edetate;

[0048] (4) 0.10-0.30% propylene glycol;

[0049] (5) 0.05-0.30% buffer;

[0050] (6) 0.0005-0.0020% antimicrobial peptides disrupt the integrity of the bacterial outer membrane and promote the penetration of subsequent antimicrobial components;

[0051] (7) 0.0001-0.0020% β-cyclodextrin-PHMB inclusion complex, where PHMB refers to polyhexamethylene hydrochloride, the molar ratio of β-cyclodextrin to PHMB is 1:2-4, and the mass ratio of PHMB to antimicrobial peptide LL-37 is 0.4-1.2:1. The free concentration of PHMB is reduced through supramolecular inclusion, and it is released into the bacteria pretreated with antimicrobial peptide LL-37.

[0052] (8) 0.001-0.005% zinc-doped mesoporous silica loaded with silver, which can be denoted as Zn-MSN(Ag) + The Zn content is 1.2-1.8 wt%, and the Ag content is... + With a loading of 0.1-0.2 wt% and a mesopore size of 4-6 nm, Zn is doped into silver-loaded mesoporous silica particles, and Zn is fixed in the SiO2 framework in a lattice-doped form. Ag + Loaded in mesoporous channels, Zn 2+ It can inhibit bacterial efflux pump genes and enhance Ag. + Intracellular retention and mesoporous confinement prolong Ag + With a release half-life of 14.7 hours or more, Zn doping can also reduce Ag... + Oxidation rate;

[0053] (9) 0.05-0.30% HEMA-dopamine liposomes, HEMA refers to hydroxyethyl methacrylate, the mass ratio of HEMA to dopamine in the liposomes is 2-4:1, the particle size is 80-120nm, HEMA-cholesterol liposomes modified with polydopamine, Zeta potential +15mV to +25mV, polydopamine anchors the hydrophobic region of the lens, HEMA chains are hydrogen bonded to tear film mucin to form a bimodal retention interface, the HPMC retention rate can reach 85% and above;

[0054] (10) The remainder is sodium hydroxide and solvent.

[0055] In this invention, the method for preparing zinc-doped mesoporous silica loaded with silver is as follows:

[0056] A sol-gel method was employed, using hexadecyltrimethylammonium bromide (CTAB) as a template, tetraethyl orthosilicate (TEOS) as the silicon source, and zinc nitrate as the zinc source. A hydrolysis-condensation reaction was carried out under alkaline conditions at 60-80℃. After the reaction, the precursor was obtained through aging, filtration, and washing. The precursor was then placed in a muffle furnace and calcined at 550-600℃ for 4-6 hours to completely remove the CTAB template agent, yielding zinc-doped mesoporous SiO2 (Zn-MSN). Subsequently, silver was loaded using a vacuum impregnation method: the Zn-MSN was placed in a vacuum dryer and evacuated to -0.1℃. The pressure was maintained at MPa for 30-60 min, and then 0.15-0.20% silver nitrate solution was slowly added under negative pressure until the material was completely submerged. After restoring to normal pressure, the material was immersed for another 12-24 h to allow silver ions to be fully adsorbed into the mesoporous channels. Finally, the loaded material was filtered, washed, and dried, and then calcined at 300-400℃ for 1-3 h to decompose the silver nitrate into silver and stably immobilize it in the mesopores, thus obtaining zinc-doped mesoporous silica loaded with silver Zn-MSN (Ag). + ).

[0057] In this invention, the preparation method of HEMA-dopamine liposomes is as follows:

[0058] Accurately weigh hydrogenated lecithin, cholesterol, and dopamine in a mass ratio of 6-8:1.5-2.5:1, and dissolve them together in an appropriate amount of chloroform / methanol mixed solvent in a volume ratio of 2:1. Stir in a water bath at 40-50℃ until completely dissolved and clear to obtain a lipid mixed solution. Then, transfer the solution to a round-bottom flask and use a rotary evaporator to remove the organic solvent by rotary evaporation under a vacuum condition of -0.08 to -0.10 MPa in a water bath at 50-60℃ until a uniform and transparent lipid film forms on the flask wall. Finally, place the round-bottom flask containing the lipid film in a vacuum drying oven and dry overnight at -0.1 MPa and 25℃ for 8-12 hours to completely remove residual solvent. Liposome preparation by hydration: Hydroxypropyl methylcellulose (HPMC) and 0.1% (w / v) hydroxyethyl methacrylate (HEMA) were dissolved together in phosphate buffer at pH 7.0-7.4 to form the inner aqueous phase; the mass ratio of hydroxyethyl methacrylate to dopamine in the inner aqueous phase was controlled at 2-4:1; the above inner aqueous phase was added to a dried lipid membrane bottle, and swollen in a water bath at 40-60℃ for 5-10 min, and then hydrated by rotating at 120-180 rpm at the same temperature for 25-35 min to completely hydrate and detach the lipid membrane, forming a crude liposome suspension; finally, the obtained suspension was extruded and granulated through 0.45 μm and 0.22 μm microporous membranes to obtain HEMA-dopamine liposomes with a particle size of 80-120 nm.

[0059] The preparation process of the rigid gas permeable contact lens solution of the present invention includes the following steps:

[0060] (1) Preliminary preparation of nursing solution:

[0061] At room temperature (20-25℃), with a low stirring speed of 200-400 rpm, the prescribed amounts of poloxamer and hydroxypropyl methylcellulose (HPMC) are sequentially dispersed in 60-70% of the total solvent volume of water for injection, and stirred until completely dissolved to form a transparent gel matrix. Then, disodium edetate, propylene glycol, and a buffer (such as BIS-TRIS) are added sequentially, and stirred until completely dissolved. The pH of the system is then adjusted to 7.0-7.6 using a 5-10% w / v sodium hydroxide solution. While maintaining stirring, the prescribed amounts of zinc-doped mesoporous silica loaded with silver, β-cyclodextrin-PHMB inclusion complex, antimicrobial peptide, and HEMA-dopamine liposomes are added sequentially, ensuring each component is evenly dispersed before adding the next. Finally, the remaining water for injection is added to the total volume, and the stirring speed is increased to 800-1000 rpm, and stirring is continued for 15-20 minutes to ensure all components are evenly mixed, yielding the initial nursing solution.

[0062] (2) Dynamic gradient sterilization treatment:

[0063] At 3-5℃, for 1200-1800s -1 Primary membrane filtration at a shear rate that protects the structural integrity of liposomes is performed, followed by three stages of pulsed moist heat sterilization:

[0064] Sterilization at 105℃ for 6-10 minutes induces liposome densification, which can increase the phase transition enthalpy by 27% or even higher.

[0065] Sterilize at 121℃ for 1-3 minutes; short-time high temperature can achieve penetration sterilization.

[0066] Sterilization at 115℃ for 4-6 minutes resulted in the formation of stable nanoparticle crystals. X-ray diffraction analysis showed that Zn-MSN(Ag) + Lattice distortion rate < 1.2%;

[0067] The standard sterilization time F0 value is ≥12min to obtain rigid gas permeable contact lens care solution.

[0068] Example 1

[0069] Step 1: Preparation of the antibacterial system

[0070] (1) Take PHMB and β-cyclodextrin in a molar ratio of 1:3, dissolve them in phosphate buffer with a pH of 7.4, and sonicate them at 45°C for 30 min to obtain β-cyclodextrin-PHMB inclusion complex.

[0071] (2) Zn-doped mesoporous silica was synthesized by sol-gel method, and then vacuum impregnated in 0.15% silver nitrate solution and calcined at 350°C for 2 hours to obtain zinc-doped mesoporous silica loaded with silver.

[0072] (3) Take hydrogenated lecithin, cholesterol and dopamine in a mass ratio of 6:2:1, mix them well and make a lipid membrane; then dissolve HPMC and 0.1% HEMA in the inner aqueous phase, and control the mass ratio of hydroxyethyl methacrylate to dopamine to be 3:1, hydrate at 50℃ for 30 min to form HEMA-dopamine liposomes with a particle size of 100±15nm.

[0073] Step 2: Prepare the nursing solution

[0074] (1) Take 3.50g poloxamer, 0.18g hydroxypropyl methylcellulose, 0.05g disodium edetate, 2.00g propylene glycol, 0.15g BIS-TRIS buffer, about 0.15g sodium hydroxide, 0.01g antimicrobial peptide LL-3, 0.0015g β-cyclodextrin-PHMB inclusion complex, and 0.03g Zn-doped mesoporous SiO2 loaded with Ag. + 1.50g HEMA-dopamine liposomes.

[0075] The weighed poloxamer and hydroxypropyl methylcellulose were dispersed in approximately 700 mL of water for injection under stirring at 400 rpm until completely dissolved, forming a transparent matrix solution. Disodium edetate, propylene glycol, and BIS-TRIS buffer were added sequentially and stirred until completely dissolved. Sodium hydroxide was dissolved in a small amount of water for injection and slowly added dropwise to the solution under stirring to adjust the pH to 7.4. Zinc-doped mesoporous silica loaded with silver, β-cyclodextrin-PHMB inclusion complex, antimicrobial peptide LL-37, and HEMA-dopamine liposomes were added sequentially, stirring for at least 5 minutes after each addition to ensure uniform dispersion. The resulting solution was transferred to a 1000 mL volumetric flask and diluted to the mark with water for injection, bringing the final volume to 1.0 L. The final solution was stirred at 800-1000 rpm for 20 minutes to ensure uniform mixing, yielding the initial care solution.

[0076] (2) Place the initial nursing solution at 4°C and heat for 1500 seconds. -1 Shear rate is used for filtering.

[0077] (3) Then perform three-stage pulsed moist heat sterilization with an F0 value ≥ 12 min. The specific process is: 105℃ for 8 min, 121℃ for 2 min, and 115℃ for 5 min to obtain the rigid corneal contact lens care solution.

[0078] Test case

[0079] (1) PHMB performance test

[0080] The rigid gas permeable contact lens solution prepared in Example 1 was used to determine the effect of polyhexamethylene hydrochloride (PHMB) on microorganisms, according to ISO 14729:2001 "Microbiological requirements and testing methods for protective products for ophthalmic optical contact lenses and hygienic management methods for contact lenses". The results are shown in Table 1 below.

[0081] Table 1 Synergistic effect of triple antibacterial system

[0082]

[0083] Wherein, *p<0.01, n=6. Specifically, the experimental data are expressed as mean ± standard deviation (Mean±SD), and the number of experimental replicates n=6 (i.e., 6 independent replicates); p<0.01 indicates that the data difference between Example 1 and the corresponding traditional PHMB control group is highly statistically significant, that is, the probability of the difference being caused by random factors is less than 1%, which can fully prove that the improvement of the antibacterial efficacy of the nursing solution of the present invention is real and reliable.

[0084] As shown in Table 1 above, the Log Reduction Value is a key indicator used in the ISO 14729 international standard to quantify antimicrobial efficacy. Its value is equal to the logarithm of the reduction in the number of microorganisms (base 10). Each increase of 1 in the Log Reduction Value represents a 90% decrease in the number of surviving microorganisms, that is, a kill rate of 90%. For example, a Log Reduction Value of 4.5 represents a kill rate of 99.997%, while a Log Reduction Value of 9.6 represents an extremely high kill rate, far exceeding the level required for disinfection and sterilization.

[0085] (2) Zn-MSN(Ag) + Performance test

[0086] 2.1 Effect of Zinc Doping on the Performance of Silver Loading

[0087] Take the Zn-MSN(Ag) prepared in Example 1 + The effect of zinc doping on the performance of silver-loaded silver was measured, using pure Ag-MSN without zinc loading as a comparison. The results are shown in Table 2 below.

[0088] Table 2 Zinc doping effect on Ag + Performance impact

[0089]

[0090] Wherein, *p<0.01 vs control group, specifically refers to the zinc-doped mesoporous silica silver-loaded [Zn-MSN(Ag)] prepared in Example 1 using the method of the present invention.+ Statistical comparison of the measurement results of the [] group and the undoped pure Ag-MSN (control group) showed that p < 0.01, indicating that the difference between the two groups was highly statistically significant (i.e., the probability of the difference being caused by random factors was less than 1%), fully demonstrating the effect of zinc doping on Ag. + The performance improvement is real and significant.

[0091] Ag + Release half-life: The significantly prolonged half-life indicates that Zn 2+ Doping inhibits Ag through mesoporous confinement effect. + The synergistic effect of migration enabled Ag + The slow, controlled release of Ag, i.e., prolonging the release of Ag + A release half-life of 14.7 hours or more helps maintain a long-lasting antibacterial concentration and avoids damage caused by Ag. + Burst release-induced cytotoxicity.

[0092] Biofilm clearance rate: The clearance rate was significantly improved, indicating that Zn 2+ Ag was enhanced by inhibiting bacterial efflux pump genes (such as mexB). + The retention and accumulation of bacteria within the microorganisms more effectively disrupts the difficult-to-remove bacterial biofilm structure, demonstrating that this invention can solve the problem of biofilm formation on lens surfaces.

[0093] Cell viability: The viability was significantly improved and maintained at an extremely high level, directly confirming that zinc doping effectively reduced Ag. + The cytotoxicity of Zn is due to its cytotoxicity. 2+ The doping stabilized Ag + This slows down its release rate and avoids high concentrations of Ag. + Instant exposure to damage corneal cells significantly improves the product's biocompatibility and safety.

[0094] As can be seen from Table 2 above, the zinc doping strategy adopted in this invention is by no means a simple mixing, but rather achieves Zn doping through lattice doping. 2+ With Ag + Functional synergy: significantly enhancing Ag + While exhibiting antibacterial efficacy, especially against biofilms, it significantly reduces cytotoxicity, perfectly resolving the industry paradox mentioned in the background technology of enhanced antibacterial properties accompanied by increased cytotoxicity.

[0095] 2.2 Zn-MSN(Ag + Performance comparison test with existing zinc-containing care solutions

[0096] Take the Zn-MSN(Ag) prepared in Example 1 +The study compared the performance of mesoporous zinc oxide and silver nitrate solutions in existing nursing care products with those in cytotoxicity, and the results are shown in Table 3 below.

[0097] Table 3 Comparison of Zn-MSN with existing zinc-containing technologies

[0098]

[0099] Wherein, *p<0.01 vs control group, specifically refers to the zinc-doped mesoporous silica silver-loaded [Zn-MSN(Ag)] prepared in Example 1 using the method of the present invention. + The measurement results of the [] group were statistically compared with those of the mesoporous zinc oxide control group and the zinc nitrate solution control group, respectively. p < 0.01 indicates that the differences between the present invention group and any control group were highly statistically significant, meaning the probability of the difference being caused by random factors was less than 1%, which sufficiently proves the Zn-MSN(Ag) of the present invention. + This technology is significantly superior to the two existing zinc-containing technologies in key performance indicators.

[0100] Zn 2+ Release rate: Zn of the present invention 2+ The release rate was significantly lower than that of the two control groups. This indicates that Zn was released through lattice doping. 2+ Zn was fixed in the SiO2 framework to realize 2+ The slow, controlled release fundamentally solves the problem of Zn mentioned in the background technology. 2+ The industry challenge of sudden release leading to corneal cytotoxicity. In contrast, mesoporous zinc oxide (Zn) 2+ The release is still relatively rapid, while the Zn in the zinc nitrate solution... 2+ These are free ions, which have the fastest release rate and the greatest potential toxicity.

[0101] Ag + Bioavailability: This invention successfully achieved Ag + Highly efficient loading capacity, while the two existing zinc-containing technologies completely lack the ability to load Ag. + This demonstrates the uniqueness and inventiveness of the zinc-doped mesoporous silica silver-supported structure created in this invention, which is not a simple combination of existing technologies. This structure enables the product to simultaneously exert the properties of Zn. 2+ With Ag + Synergistic antibacterial effect.

[0102] Corneal cell survival rate: The corneal cell survival rate of this invention was significantly higher than that of the two control groups, reaching as high as 96.3%, demonstrating its excellent biocompatibility and extremely low cytotoxicity. This is attributed to the combined effects of Zn. 2+Its sustained-release properties prevent burst ion release, and Ag + High-efficiency load and controllable release. In contrast, existing technologies suffer from high Zn content. 2+ Released too quickly or unable to avoid Ag + The toxicity of the cells leads to low cell survival rates.

[0103] As can be seen from Table 3 above, the Zn-MSN(Ag) developed in this invention... + This technology surpasses existing zinc-containing technologies in core performance. It doesn't simply mix zinc and silver; instead, through innovative material structure design, it simultaneously achieves the desired performance for both zinc and silver. 2+ Sustained release, Ag + With three major improvements—high loading capacity and significantly reduced cytotoxicity—a highly efficient and safe multifunctional antibacterial agent has been successfully developed, providing a better solution for rigid gas permeable contact lens care solutions.

[0104] 2.3 Flow cytometry was used to detect fluorescently labeled PHMB.

[0105] Take a suspension of Staphylococcus aureus (ATCC 6538) in the logarithmic growth phase (1×10⁻⁶). 8 The bacterial culture was divided into two groups (CFU / mL). The experimental group was pretreated with 0.0005% (w / v) of the antimicrobial peptide LL-37 for 10 min, while the control group was treated with buffer. Both groups were then incubated with FITC-labeled β-CD / PHMB inclusion complex (final concentration 0.0005%) for 30 min in the dark, followed by centrifugation and washing to remove any fluorescent markers that did not enter the bacterial cells. The mean fluorescence intensity (MFI) of 10,000 bacteria was detected by flow cytometry.

[0106] The results showed that the MFI of the control group was 52.3 ± 4.1, while the MFI of the experimental group (LL-37 pretreatment) was 220.5 ± 12.8. The MFI of the experimental group was 4.22 times that of the control group. Calculations showed that the membrane perforation effect of LL-37 increased the efficiency of β-CD / PHMB entering the bacterial cells by approximately 3.2 times.

[0107] 2.4 The relative expression level of the mexB gene was detected using qPCR technology.

[0108] Take a suspension of *Pseudomonas aeruginosa* (ATCC 27853) in the logarithmic growth phase and dilute it with 0.002% (w / v) pure Ag-MSN and Zn-MSN (Ag... + The bacteria were treated for 2 hours. After treatment, total bacterial RNA was extracted using an RNA extraction kit and reverse transcribed into cDNA. The relative expression level of the mexB gene was detected using qPCR with 16S rRNA as an internal reference gene.

[0109] Primer sequences:

[0110] mexB-F:5'-ATCGGCTACTTCGAGCAGAC-3',

[0111] mexB-R:5'-CGTCTTCAGCATAGCCAGGT-3'.

[0112] The results showed that, compared with the untreated group (expression level set at 100%), the expression level of the mexB gene in the pure Ag-MSN treatment group was 105.4% ± 8.7%, which was not significantly different from the control group (p>0.05). Meanwhile, the expression level of Zn-MSN (Ag... + The expression level of mexB gene in the treatment group decreased to 22.1%±3.5%, which is a decrease of 77.9% (p<0.01).

[0113] 2.5 Measurement of polydopamine coating using atomic force microscopy

[0114] Rigid contact lens material sheets were fixed onto the sample stage of an atomic force microscope (AFM). Modified AFM probes were used to simulate unmodified liposomes and polydopamine-modified HEMA-liposomes, respectively. In a simulated tear environment, the adhesion force and adhesion work during probe separation from the material surface were measured using AFM force curve techniques.

[0115] The results showed that the average adhesion work of unmodified liposomes was -22.4 ± 3.1 mJ / m 2 The average adhesion work of polydopamine-modified HEMA liposomes was -38.6 ± 2.8 mJ / m. 2 The larger the absolute value of the adhesion work (negative value), the stronger the adsorption energy, proving that the polydopamine coating significantly improves the adsorption energy of liposomes, thus achieving the long-lasting lubrication mentioned above.

[0116] 2.6 PHMB degradation rate was determined by HPLC coupled with dynamic light scattering.

[0117] Samples of the care solutions that underwent gradient sterilization and conventional sterilization at 121°C were taken. The characteristic peak area of ​​PHMB was detected using HPLC with a C18 column. The residual rate was calculated using an unsterilized initial care solution as a control: Degradation rate = (1 - Residual rate) × 100%.

[0118] Liposome encapsulation efficiency (dynamic light scattering DLS): The particle size distribution of HEMA-dopamine liposomes in the care solution before and after sterilization was measured using a dynamic light scattering instrument. The encapsulation efficiency after sterilization was calculated by measuring the content of free HPMC.

[0119] The results showed that the residual rate of PHMB after gradient sterilization was 96.8%±1.2%, which is a degradation rate of 3.2% (<3.5%), while the residual rate of PHMB after traditional sterilization was 82.5%±3.5%. The encapsulation rate of liposomes after gradient sterilization was 93.5%±1.8% (>92%), while the encapsulation rate after traditional sterilization was 75.4%±4.2%. The data prove that the gradient sterilization process can effectively sterilize while greatly protecting the stability of active ingredients.

[0120] 2.7 PHMB cytotoxicity was detected using the CCK-8 assay.

[0121] The median lethal concentration (LD50) of different concentrations of free PHMB and β-CD / PHMB inclusion complex against human corneal epithelial cells (HCECs) was determined using the CCK-8 assay. Eye irritation (rabbit eye test): Following the Draize test criteria, free PHMB (0.005%) and the PHMB-containing solution of this invention were instilled into the eyes of New Zealand rabbits. The reactions of the conjunctiva, cornea, and iris were observed and scored at 1 h, 24 h, 48 h, and 72 h.

[0122] The results showed that the LD50 of free PHMB was 0.005%, while the LD50 of the β-CD / PHMB inclusion complex increased to 0.032%, and the cytotoxicity decreased to 1 / 6.4 of the original formulation. The Draize score of the free PHMB group was 1.9, which is considered mild irritation. The Draize score of the care solution group of this invention was 0.3, which is within the non-irritating range, confirming that the inclusion technology significantly reduced irritation.

[0123] The above are merely preferred embodiments of the present invention and are not intended to limit the present 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 rigid gas permeable contact lens care solution, characterized in that, It includes a multiple antibacterial system; the multiple antibacterial system includes antimicrobial peptides, β-cyclodextrin-PHMB inclusion complex, zinc-doped mesoporous silica loaded with silver and HEMA-dopamine liposomes; in the zinc-doped mesoporous silica loaded with silver, the mesoporous silica serves as the framework material, zinc is doped into the mesoporous silica framework material in the form of lattice doping, and silver is loaded into the mesoporous channels.

2. The rigid gas permeable contact lens solution according to claim 1, characterized in that, The method for preparing the zinc-doped mesoporous silica loaded with silver is as follows: zinc-doped mesoporous silica is synthesized by sol-gel method, and then loaded with silver nitrate solution by vacuum impregnation; then calcination is performed to obtain the zinc-doped mesoporous silica loaded with silver.

3. The rigid gas permeable contact lens solution according to claim 2, characterized in that, In the method for preparing zinc-doped mesoporous silica loaded with silver, the calcination temperature is 300-400℃ and the calcination time is 1-3h.

4. The rigid gas permeable contact lens solution according to claim 1, characterized in that, In the zinc-doped mesoporous silica loaded with silver, the zinc content is 1.2-1.8% by mass and the silver loading is 0.1-0.2 wt%.

5. The rigid gas permeable contact lens solution according to claim 1, characterized in that, In the zinc-doped mesoporous silicon dioxide loaded with silver, the mesopore diameter is 4-6 nm.

6. The rigid gas permeable contact lens solution according to claim 1, characterized in that, In the β-cyclodextrin-PHMB inclusion complex, the molar ratio of β-cyclodextrin to PHMB is 1:2-4, and the mass ratio of PHMB to antimicrobial peptide is 0.4-1.2:

1.

7. The rigid gas permeable contact lens solution according to claim 1, characterized in that, In the HEMA-dopamine liposomes, the mass ratio of HEMA to dopamine is 2-4:1, and the particle size of the HEMA-dopamine liposomes is 80-120 nm.

8. A preparation process for the rigid gas permeable contact lens care solution according to any one of claims 1 to 7, characterized in that, The process includes the following steps: S1 Preliminary preparation of the contact lens solution: Take raw materials containing a multi-antibacterial system, mix them well, and obtain the preliminary contact lens solution; S2 Filtration treatment: Filter the preliminary contact lens solution through a membrane filter; S3 Gradient sterilization treatment: Perform the following three-stage pulsed moist heat sterilization in sequence: sterilize at 105℃ for 6-10 min, sterilize at 121℃ for 1-3 min, and sterilize at 115℃ for 4-6 min to obtain the rigid gas permeable contact lens solution.

9. The preparation process of the rigid gas permeable contact lens solution according to claim 8, characterized in that, In step S1, the raw materials, by mass concentration, include: 0.01-0.70% poloxamer; 0.01-0.35% hydroxypropyl methylcellulose; 0.01-0.10% disodium edetate; 0.10-0.30% propylene glycol; 0.05-0.30% buffer; 0.0005-0.0020% antimicrobial peptide; 0.0001-0.0020% β-cyclodextrin-PHMB inclusion complex; 0.001-0.005% zinc-doped mesoporous silica loaded with silver; 0.05-0.30% HEMA-dopamine liposomes; the balance being sodium hydroxide and solvent.

10. The preparation process of the rigid gas permeable contact lens solution according to claim 8, characterized in that, In step S2, during membrane filtration, the temperature is controlled at 3-5℃ and the shear rate is 1200-1800 s. -1 .

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