阅读说明：本技术 一种载药抗黏附隐形眼镜水凝胶材料及制备方法 (Drug-loaded anti-adhesion contact lens hydrogel material and preparation method thereof ) 是由 张静 陈凌东 于 2020-12-29 设计创作，主要内容包括：本发明涉及高分子水凝胶技术领域,尤其涉及一种载药抗黏附隐形眼镜水凝胶材料及制备方法,所述载药抗黏附隐形眼镜水凝胶材料由甲基丙烯酸羟乙酯、丙烯酸、两性离子单体与疏水单体共聚后负载药物而成。本发明的载药抗黏附隐形眼镜水凝胶材料具有良好的透光度、一定的载药能力并能实现药物的缓释、稳定的脱水-补液性能、较高的含水量和稳定的保水率、优异的抗蛋白、抗细菌黏附性能；制备方法简洁、高效且环保,对设备无特殊要求,对环境友好,易于产业化。(The invention relates to the technical field of high-molecular hydrogel, in particular to a drug-loaded anti-adhesion contact lens hydrogel material and a preparation method thereof. The drug-loaded anti-adhesion contact lens hydrogel material has good transmittance and certain drug-loading capacity, can realize the slow release and stable dehydration-fluid infusion performance of the drug, has higher water content and stable water retention rate, and has excellent anti-protein and anti-bacterial adhesion performance; the preparation method is simple, efficient, environment-friendly, free of special requirements for equipment, environment-friendly and easy to industrialize.)

1. The drug-loaded anti-adhesion contact lens hydrogel material is characterized by being prepared by copolymerizing hydroxyethyl methacrylate, an acrylic monomer, a zwitterionic monomer and a hydrophobic monomer and then loading a drug.

2. The drug-loaded anti-adhesion contact lens hydrogel material as claimed in claim 1,

the zwitterionic monomer comprises sulfobetaine methyl methacrylate or carboxylic betaine methyl methacrylate;

the hydrophobic monomer comprises methyl methacrylate;

the drug is norfloxacin or samarol.

3. The drug-loaded anti-adhesion contact lens hydrogel material according to claim 1, wherein the maximum transmittance of the drug-loaded anti-adhesion contact lens hydrogel material is 95%.

4. A method for preparing a drug-loaded anti-adhesive contact lens hydrogel material according to any one of claims 1 to 3, comprising the steps of:

(1) preparing a mixed solution of hydroxyethyl methacrylate, acrylic acid, a zwitterionic monomer, a hydrophobic monomer, a cross-linking agent and an initiator;

(2) deoxidizing the mixed solution, and removing bubbles in the mixed solution to obtain a pre-solution;

(3) sealing the pre-solution in a light-transmitting mold, and carrying out ultraviolet irradiation reaction to obtain hydrogel;

(4) and (3) freeze-drying the hydrogel, and then immersing the hydrogel into a medicinal solution until the hydrogel is balanced in swelling, thereby obtaining the medicine-carrying anti-adhesion contact lens hydrogel material.

5. The preparation method according to claim 4, wherein in the step (1), the mass percentage concentration of each component in the mixed solution is as follows: 5-40 wt% of hydroxyethyl methacrylate, 5-40 wt% of zwitterionic monomer, 0.5-2.5 wt% of acrylic monomer, 2-10 wt% of hydrophobic monomer, 0.2-2 wt% of cross-linking agent and 0.2-1 wt% of initiator.

6. The method according to claim 4, wherein in the step (1), the initiator is 2-hydroxy-4- (2-hydroxyethoxy) -2-methylpropiophenone.

7. The method according to claim 4, wherein in the step (1), the crosslinking agent is polyethylene glycol dimethacrylate having a molecular weight of 550 or 750.

8. The preparation method according to claim 4, wherein in the step (2), the oxygen removal is performed by introducing nitrogen or inert gas to reduce oxygen solubility; the method for removing the bubbles in the mixed solution is ultrasonic.

9. The production method according to claim 4, wherein in the step (3): the light-transmitting mold comprises a glass mold; the ultraviolet light with the wavelength of 365nm is selected during the ultraviolet light irradiation reaction; the reaction time of the ultraviolet irradiation is 3-4 h.

10. The preparation method according to claim 4, wherein in the step (4), the drug is norfloxacin or samarol, and the concentration of the drug solution is 240-260 μ g/mL.

Technical Field

The invention relates to the technical field of high-molecular hydrogel, in particular to a drug-loaded anti-adhesion contact lens hydrogel material and a preparation method thereof.

Background

A hydrogel is a material in which hydrophilic polymers are crosslinked by physical interaction or chemical reaction to form a three-dimensional network structure, and can be sufficiently swollen in water without being dissolved. In recent years, hydrogel has been widely used in various fields, such as contact lens materials, tissue engineering, wound dressings, seawater desalination, and the like.

In practical application, the hydrogel can still keep stable performance after multiple dehydration-fluid infusion cycles, and the method has important significance. For example, the contact lens material may be dehydrated due to evaporation of water during wearing and use, and may absorb water when re-immersed in the care solution, and the liquid replacement capability may be reduced during this process, so that the properties of the contact lens material, such as shape, thickness, curvature, etc., may be changed, and the subsequent use may be affected. Therefore, it is important to prepare gel materials with controlled wetting and swelling properties.

The bioavailability of common commercial eye drops is often less than 10% due to blinking and tear clearance. After the contact lens is loaded with the drug, the contact lens can be used as a drug delivery system for treating chronic and acute diseases such as glaucoma. The hydrogel has good swelling performance, can absorb a drug solution to achieve the effect of loading the drug, increases the contact time of the drug on the surface of the cornea, and can improve the bioavailability of the drug. The drug is further combined with the contact lens material through a special acting force, so that the loading capacity and the stable release capacity of the drug can be improved.

During the wearing and using process of the contact lens, protein secreted by eyes and the adhesion of external bacteria on the surface of the material are important factors influencing the service life of the contact lens and the sanitation and safety of the eyes. Zwitterionic materials are widely used in biomedical and engineering applications because of their excellent antifouling properties. The molecular chain of the zwitterionic polymer is simultaneously provided with anionic groups and cationic groups, and the zwitterionic polymer can be strongly combined with water molecules through solvation to generate repulsive force, so that the adhesion of proteins or bacteria and cells can be effectively resisted.

Disclosure of Invention

In order to overcome the problems in the prior art, the invention provides a drug-loaded anti-adhesion contact lens hydrogel material which has stable dehydration-fluid infusion performance, excellent anti-protein and anti-bacterial adhesion performance and good light transmittance, and the hydrogel has certain drug loading capacity and can realize the slow release of drugs.

The invention also provides a preparation method of the drug-loaded anti-adhesion contact lens hydrogel material, and the method is simple, efficient and environment-friendly, has no special requirements on equipment, is environment-friendly and is easy to industrialize.

In order to achieve the purpose, the invention adopts the following technical scheme:

the drug-loaded anti-adhesion contact lens hydrogel material is prepared by copolymerizing hydroxyethyl methacrylate, an acrylic monomer, a zwitterionic monomer and a hydrophobic monomer and then loading a drug. The hydroxyl group in the hydroxyethyl methacrylate endows the hydrogel with better hydrophilicity and generates a hydrogen bond with the norfloxacin monomer to enhance the drug loading capacity, the carboxyl group in the acrylic acid and the amino group of the norfloxacin monomer generate a static acting force to endow the hydrogel with good drug loading capacity, the zwitter ion part endows the hydrogel with excellent anti-adhesion performance and improves the solubility of the hydrophobic monomer in the pre-solution, and the hydrophobic monomer is used for adjusting the swelling performance of the hydrogel.

Preferably, the zwitterionic monomer is sulfobetaine methyl methacrylate (SBMA) or carboxylic betaine methyl methacrylate (CBMA); preferably sulfobetaine methyl methacrylate (SBMA).

The hydrophobic monomer is Methyl Methacrylate (MMA);

the drug is norfloxacin or samarol.

Preferably, the maximum transmittance of the drug-loaded anti-adhesion contact lens hydrogel material is 95%, and the drug-loaded anti-adhesion contact lens hydrogel material has high water content, stable water retention rate, high drug-loading capacity and excellent anti-protein and anti-bacterial adhesion properties.

A preparation method of a drug-loaded anti-adhesion contact lens hydrogel material comprises the following steps:

(1) preparing a mixed solution of hydroxyethyl methacrylate, acrylic acid, a zwitterionic monomer, a hydrophobic monomer, a cross-linking agent and an initiator;

(2) deoxidizing the mixed solution, and removing bubbles in the mixed solution to obtain a pre-solution;

(3) sealing the pre-solution in a light-transmitting mold, and carrying out ultraviolet irradiation reaction to obtain hydrogel;

(4) and (3) freeze-drying the hydrogel, and then immersing the hydrogel into a medicinal solution until the hydrogel is balanced in swelling, thereby obtaining the medicine-carrying anti-adhesion contact lens hydrogel material.

Preferably, in the step (1), the mass percentage concentration of each component in the mixed solution is as follows: 5-40 wt% of hydroxyethyl methacrylate, 5-40 wt% of zwitterionic monomer, 0.5-2.5 wt% of acrylic monomer, 2-10 wt% of hydrophobic monomer, 0.2-2 wt% of cross-linking agent and 0.2-1 wt% of initiator. Dissolving hydroxyethyl methacrylate, acrylic acid, a zwitterionic monomer, a hydrophobic monomer, a cross-linking agent and an initiator in a solvent, and then uniformly stirring; the solvent includes water.

Preferably, in the step (1), the initiator is 2-hydroxy-4- (2-hydroxyethoxy) -2-methyl propiophenone (I2959), and the I2959 endows the hydrogel monomer with a higher initiation rate.

Preferably, in the step (1), the crosslinking agent is polyethylene glycol dimethacrylate with molecular weight of 550 or 750; polyethylene glycol dimethacrylate having a molecular weight of 550 is preferred.

Preferably, in the step (2), the oxygen removal is performed by introducing nitrogen or inert gas to reduce oxygen solubility; the method for removing the bubbles in the mixed solution is ultrasonic.

Preferably, in step (3): the light-transmitting mold comprises a glass mold; the ultraviolet light with the wavelength of 365nm is selected during the ultraviolet light irradiation reaction; the duration of the ultraviolet irradiation reaction is 3-4 hours, preferably 3 hours.

Preferably, in the step (4), the drug is norfloxacin or samarol, and the concentration of the drug solution is 240-260 μ g/mL, and more preferably 250 μ g/mL.

Therefore, the invention has the following beneficial effects:

(1) the drug-loaded anti-adhesion contact lens hydrogel material has good transmittance and certain drug-loading capacity, can realize the slow release and stable dehydration-fluid infusion performance of the drug, has higher water content and stable water retention rate, and has excellent anti-protein and anti-bacterial adhesion performance;

(2) the preparation method is simple, efficient, environment-friendly, free of special requirements for equipment, environment-friendly and easy to industrialize.

Drawings

FIG. 1 shows the transmittance of hydrogels prepared in examples 4 to 7 of the present invention;

FIG. 2 shows the swelling properties of the hydrogels prepared in examples 4 to 7 of the present invention;

FIG. 3 is a graph showing the mass change rate of the hydrogel repeatedly dehydrated and swollen (swollen in deionized water) in examples 4 to 7 of the present invention;

FIG. 4 is a graph showing the water content and water retention of hydrogels obtained in examples 4-7 of the present invention (A is the water content of the hydrogels obtained in examples 4-7 at room temperature (swollen in deionized water), B is the water retention of the hydrogels obtained in examples 4-7 at room temperature (swollen in deionized water), C is the water content of the hydrogels obtained in examples 4-7 at room temperature (swollen in a commercially available care solution), and D is the water retention of the hydrogels obtained in examples 4-7 at room temperature (swollen in a commercially available care solution);

FIG. 5 shows the cell compatibility of the hydrogel prepared in example 4 of the present invention;

FIG. 6 shows the drug loading of the hydrogels prepared in examples 4 to 7 of the present invention;

FIG. 7 is a drug release curve of the hydrogel prepared in examples 4 to 7 of the present invention;

FIG. 8 shows the protein adhesion resistance of the hydrogels prepared in examples 4 to 7 of the present invention to commercially available contact lenses;

FIG. 9 shows the anti-E.coli adhesion of the hydrogels prepared in examples 4-7 of the present invention and commercially available contact lenses.

Detailed Description

The technical solution of the present invention is further specifically described below by using specific embodiments and with reference to the accompanying drawings.

In the present invention, all the equipment and materials are commercially available or commonly used in the art, and the methods in the following examples are conventional in the art unless otherwise specified.

Examples 1 to 9

A preparation method of a novel medicine-carrying anti-adhesion hydrogel capable of being used as a contact lens comprises the following preparation steps:

1) preparing a mixed solution of hydroxyethyl methacrylate, acrylic acid, a zwitterionic monomer, a hydrophobic monomer, a cross-linking agent and an initiator;

2) deoxidizing the mixed solution, and removing bubbles in the mixed solution by ultrasonic waves to obtain a pre-solution;

3) sealing the pre-solution in a light-transmitting glass mold, and carrying out ultraviolet irradiation reaction for 5 hours at the wavelength of 365nm to prepare hydrogel;

4) and (3) freeze-drying the hydrogel, and then immersing the hydrogel into a drug solution until the swelling is balanced to obtain the novel drug-loaded anti-adhesion hydrogel.

Wherein, the initiator in the step 1) is I2959, the cross-linking agent is PEGDMA (Mn 550), and all solvents of the mixed solution are deionized water; the zwitterionic monomer is SBMA, the hydrophobic monomer is MMA, and the drug is norfloxacin.

Step 1) the ingredients are shown in table 1 below.

Table 1 ingredient table for examples 1-9.

The hydrogel is prepared according to the ingredients in table 1 and the preparation parameters in table 2, and the hydrogel prepared in the step (3) in the embodiment 1-9 is subjected to performance test after the preparation is finished.

The performance test comprises the following aspects:

(1) and (3) testing the transmittance: a circular hydrogel sample having a diameter of 2.5cm was prepared, and the light transmittance of the sample was measured using an ultraviolet/visible/near-infrared spectrophotometer.

(2) Dehydration-fluid infusion test: placing the hydrogel sheet sample in an oven at 50 ℃ for 1h, weighing and recording as W₁(ii) a The weighed hydrogel sample was then placed in deionized water to swelling equilibrium and weighed as W₂. The above steps are repeated for seven cycles. The original hydrogel sample mass is W₀. Each sample was tested in triplicate and the average was taken. The hydrogel mass change rate was calculated according to the following equation.

(3) Testing the water content and the water retention rate: placing the hydrogel sample at room temperature, weighing and recording the hydrogel sample after being placed for different time as W_tThe completely dried hydrogel sample was weighed as W_d. The hydrogel mass at swelling equilibrium is recorded as W_s. The water content and water retention of the hydrogel were determined according to the following formulas.

(4) Drug loading and drug release test: the lyophilized hydrogel was soaked in 250. mu.g/mL norfloxacin solution and protected from light overnight. The absorbance of the drug solution before and after the soaking at 273nm was measured with a microplate reader (DG5033A), and the drug loading of the hydrogel was determined. Soaking the norfloxacin-loaded hydrogel into 10mL of phosphate buffer solution, placing the norfloxacin-loaded hydrogel in a water bath kettle at 37 ℃, taking out 1mL of liquid at intervals, and measuring the absorbance at 273nm by using an enzyme-labeling instrument to obtain the drug release curve of the hydrogel.

(5) And (3) testing the protein adhesion resistance: the swollen and sterilized hydrogel sheet-like sample was placed in a 24-well plate, 1mL of goat anti-human globulin (HRP-IgG) labeled with horseradish peroxidase was added to each well, and after 12 hours at 37 ℃ in an incubator, a portion of the sample was sonicated in PBS buffer for 1 min. The treated sample was placed in a new 24-well plate, 1mL of 0.1M citrate-phosphate buffer (pH 5) containing 1 μ g/mL o-phenylenediamine and 0.03% hydrogen peroxide was added to the well, and after 15min 2mL of 2M H was added₂SO₄The reaction was terminated. The supernatant was used to determine the Optical Density (OD) at 492nm using a microplate reader, and the results were the average of three experiments. Another portion of the sample was sonicated in a commercial contact lens solution for 1min, and fresh protein solution was added again, and the above procedure was repeated five times.

(6) And (3) testing antibacterial performance: placing the swollen and sterilized hydrogel sheet sample in a 12-hole plate, adding 1mL of escherichia coli bacterial liquid with Optical Density (OD) of 0.1 into each hole, then placing the 12-hole plate in a shaking table, setting the temperature at 37 ℃ and the rotating speed at 120rpm, and co-culturing the sample and the bacterial liquid for 24 h. After the co-culture is finished, taking out the sample from the bacterial liquid, carrying out ultrasonic treatment in a PBS buffer solution for 1min, dyeing for 10min in a dark environment by using a dye, and observing the adhesion condition of bacteria on the surface of the sample by using an inverted fluorescence microscope.

(7) In vitro cytotoxicity experiments: l929 cells were seeded in 96-well plates at a density of 5X 10 per well⁴And cultured at 37 ℃ for 24 hours. Then, the cell culture medium was replaced with 200 μ L of hydrogel percolate with different concentrations and the cells were cultured for further 24 h. Subsequently, the medium was removed and 200. mu.L of MTT solution (0.5mg/mL) was added. After 4h of reaction, the medium was replaced with 150. mu.L of dimethyl sulfoxide. Finally, the well plate was placed in a microplate reader (DG5033A) and the absorbance of the solution in the well was measured at 492 nm. The results were defined as the average percentage of cell viability relative to untreated cells and were calculated as follows

Obtaining OD in the Presence of hydrogel leachate_treatedObtaining OD in the absence of hydrogel leachate_controlAnd obtaining the OD in the absence of hydrogel permeant and cells_blank. OD values were measured four times on separate parallel samples.

The transmittance of the hydrogels prepared in examples 4-7 is shown in FIG. 1, where the hydrogel prepared according to the formulation described in example 7 has the highest transmittance, greater than 95% in the visible wavelength.

The swelling curves of the hydrogels prepared in examples 4 to 7 are shown in fig. 2, and the swelling ratios of the hydrogels prepared by the formulations described in examples 4 to 7 are 19.95%, 58.76%, 87.42%, and 121.24%, respectively.

The mass change rate of the hydrogel prepared in the examples 4 to 7 after repeated dehydration and fluid infusion in deionized water is shown in fig. 3, and the hydrogel prepared by the formulations in the examples 4 to 7 has stable dehydration and fluid infusion capability.

The water content and the water retention of the hydrogel prepared in the embodiments 4 to 7 are shown in fig. 4, the water content and the water retention of the hydrogel increase with the increase of the SBMA content, and the hydrogel prepared by the formula in the embodiment 7 has the highest water content and the strongest water retention capacity.

The cell compatibility of the hydrogel prepared in example 4 is shown in FIG. 5, and the cell survival rate is higher than 85% after the hydrogel is cultured with different concentrations of hydrogel leaching solution for different time.

The drug loading capacity of the hydrogel prepared in examples 4 to 7 is shown in fig. 6, wherein the hydrogel prepared in example 7 has the highest drug loading capacity of 11.66 mg/mL.

The release curves of the hydrogels prepared in examples 4 to 7 are shown in FIG. 7, and the hydrogels prepared in examples 4 to 7 all have stable release capacity.

The protein adhesion resistance of the hydrogels obtained in examples 4 to 7 is shown in fig. 8, and the hydrogel obtained in example 7 has the lowest protein adsorption rate, which is lower than that of the surface of the commercially available contact lens in all 7 cycles.

The adhesion of the hydrogels prepared in examples 4 to 7 to E.coli is shown in FIG. 9, and the hydrogels prepared in examples 4 to 7 all have excellent adhesion to E.coli.

A large number of experimental results show that the medicine-carrying anti-adhesion hydrogel prepared by the technical scheme of the invention really has good transmittance and certain medicine-carrying capacity, can realize slow release of medicines, stable dehydration-fluid infusion performance, higher water content, stable water retention rate and excellent anti-protein and anti-bacterial adhesion performance, and can be used as an ideal material for preparing contact lenses.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and other variations and modifications may be made without departing from the spirit of the invention as set forth in the claims.