An ophthalmic liquid composition, its method of preparation and use

Abstract

The application provides an eye liquid composition and a preparation method and application thereof, and belongs to the technical field of eye care. The eye liquid composition provided by the application comprises 0.0001-0.002% of punicalagin in mass content. The application introduces a proper amount of the natural polyphenol compound punicalagin with broad-spectrum and high-efficiency antibacterial activity into the eye liquid composition, so that the dosage of the traditional chemical preservative is greatly reduced, and even the chemical preservative can be omitted, the cytotoxicity and irritability to the ocular surface are reduced, and the safety is improved. In addition, the punicalagin can be quickly and uniformly adsorbed on the surface of a contact lens, and is slowly released during wearing, so as to provide sustained antibacterial, anti-inflammatory and antioxidant protection for the ocular surface.

Description

Technical Field

[0001] This invention belongs to the field of eye care technology, specifically relating to an eye liquid composition, its preparation method, and its application. Background Technology

[0002] Eye care products, such as contact lens care products and eye washes, often use chemical preservatives as their main antibacterial ingredients. These chemical preservatives mainly include: biguanide preservatives, represented by polyhexamethylene biguanide (PHMB); and quaternary ammonium salt preservatives, represented by polyquaternium-1 (POLYQUAD). The problem with these chemical preservatives is that, at effective antibacterial concentrations, they all exhibit varying degrees of ocular surface toxicity, irritation, or allergenicity; while reducing the concentration to ensure safety often results in insufficient antibacterial efficacy.

[0003] Therefore, there is an urgent need for an ophthalmic liquid composition with strong antibacterial activity and high ocular surface safety. Summary of the Invention

[0004] The purpose of this invention is to provide an ophthalmic liquid composition, its preparation method, and its application. The ophthalmic liquid composition provided by this invention exhibits strong antibacterial activity and high safety for the ocular surface.

[0005] To achieve the above-mentioned objective, the present invention provides an ophthalmic liquid composition comprising 0.0001 to 0.002% by mass of pungent glycoside.

[0006] Preferably, it includes 0.0005 to 0.001% pungent glycoside by mass content.

[0007] Preferably, by mass content, it also includes 0.005~0.1% of a moisturizing lubricant, 0.01~0.5% of a buffer, 0.1~0.5% of an osmotic pressure regulator, and the balance being purified water.

[0008] Preferably, by mass content, it further includes 0.005~0.1% of moisturizing lubricant, 0.01~0.5% of buffer, 0.1~0.5% of osmotic pressure regulator, 0.025~0.1% of ion chelating agent, 0~0.000025% of preservative and the balance being purified water.

[0009] Preferably, when the preservative content is not 0, the mass ratio of the preservative to pungent glycoside is 1:(20~100).

[0010] Preferably, the preservative is one or more of polyhexamethylene biguanide, sorbic acid, polyquaternium salt and polyaminopropyl biguanide.

[0011] Preferably, the ion chelating agent is disodium ethylenediaminetetraacetate and / or sodium citrate.

[0012] The present invention also provides a method for preparing the ophthalmic liquid composition described above, comprising: mixing punicin and a portion of purified water under light-protected conditions, then adding a moisturizing lubricant, a buffer, an osmotic pressure regulator and the remaining purified water for a second mixing, followed by filtration to obtain the ophthalmic liquid composition; Alternatively, it may include: under light-protected conditions, first mixing pungent glycoside and a portion of purified water, then adding a moisturizing lubricant, a buffer, an osmotic pressure regulator, an ion chelating agent, a preservative, and the remaining purified water for a second mixing, followed by filtration to obtain an ophthalmic liquid composition.

[0013] The present invention also provides the application of the ophthalmic liquid composition described in the above technical solution or the ophthalmic liquid composition prepared by the preparation method described in the above technical solution in eye drops, eye rinse solutions and eye wash solutions.

[0014] The present invention also provides the application of the ophthalmic liquid composition described in the above-described technical solutions or the ophthalmic liquid composition prepared by the preparation method described in the above-described technical solutions in eyeglass preservation solutions and contact lens care solutions.

[0015] This invention provides an ophthalmic liquid composition comprising 0.0001-0.002% pungent glycoside by mass. By introducing an appropriate amount of pungent glycoside, a natural polyphenol compound with broad-spectrum and highly effective antibacterial activity, into the ophthalmic liquid composition, this invention significantly reduces the amount of traditional chemical preservatives used, and may even eliminate the need for chemical preservatives altogether. This reduces toxicity and irritation to ocular surface cells, thereby improving safety. Furthermore, pungent glycoside can be rapidly and uniformly adsorbed onto the surface of contact lenses and slowly released during wear, providing continuous antibacterial, anti-inflammatory, and antioxidant protection to the ocular surface. Attached Figure Description

[0016] Figure 1 The initial state photograph of the lens was selected when testing the lens compatibility of the contact lens solution prepared in Example 1 of this invention in a silicone hydrogel lens. Figure 2 Photographs of the state of the contact lens after immersion for 7 days in the contact lens compatibility test of the contact lens solution prepared in Example 1 of this invention in a silicone hydrogel lens. Figure 3 The adsorption kinetics curve of punicalin in the contact lens care solution prepared in Example 1 of this invention on a silicone hydrogel lens is shown. Figure 4 The release curve of pungent glycoside from silicone hydrogel lenses to artificial tears in the contact lens care solution prepared in Test Example 1 of this invention is shown. Detailed Implementation

[0017] The present invention provides an ophthalmic liquid composition comprising 0.0001 to 0.002% by mass of pungent glycoside.

[0018] The ophthalmic liquid composition provided by this invention includes 0.0001~0.002% punicalin by mass, preferably 0.0005~0.001% punicalin by mass. As one embodiment of this invention, it may contain 0.0001% punicalin by mass, or 0.0003% punicalin by mass, or 0.0005% punicalin by mass, or 0.001% punicalin by mass, or 0.002% punicalin by mass. Punicalin is the main component of pomegranate peel polyphenols, possessing broad-spectrum antibacterial activity, antioxidant and anti-inflammatory activity, and good safety. The addition of the above-mentioned punicalin by mass content in the ophthalmic liquid composition ensures the exertion of antibacterial, anti-inflammatory, and antioxidant effects, while also providing good thermal stability, suitable for storage and processing at room temperature or low temperature.

[0019] In an embodiment of the present invention, the manufacturer of pungent glycoside is MCE, with catalog number HY-N0063.

[0020] In one embodiment of the present invention, the ophthalmic liquid composition may further include, by mass content, 0.005-0.1% of a moisturizing lubricant, 0.01-0.5% of a buffer, 0.1-0.5% of an osmotic pressure regulator, and the balance being purified water.

[0021] In one embodiment of the present invention, the mass content of the moisturizing lubricant can be 0.005~0.01%, or 0.01~0.1%; the moisturizing lubricant can be any one or more of hydroxypropyl methylcellulose, trehalose, sodium hyaluronate, propylene glycol, polyethylene glycol, sodium carboxymethyl cellulose, and ectoine. The addition of the above-mentioned mass content of moisturizing lubricant can achieve the functions of lubrication, friction reduction, water reduction, and anti-drying.

[0022] In one embodiment of the present invention, the mass content of the buffer can be 0.01~0.1% or 0.1~0.5%; the buffer can be one or more of sodium dihydrogen phosphate and disodium hydrogen phosphate, tromethamine and hydrochloric acid, boric acid and borax. The addition of the above buffer can prevent corneal burns caused by excessive acidity or alkali.

[0023] In one embodiment of the present invention, the mass content of the osmotic pressure regulator can be 0.1~0.3% or 0.3~0.5%; the osmotic pressure regulator can be sodium chloride and / or potassium chloride. The addition of the above-mentioned mass content of the osmotic pressure regulator can match the physiological osmotic pressure of human tears.

[0024] In another technical solution of the present invention, as an embodiment, the ophthalmic liquid composition may further include, by mass content, 0.005~0.1% of a moisturizing lubricant, 0.01~0.5% of a buffer, 0.1~0.5% of an osmotic pressure regulator, 0.025~0.1% of an ion chelating agent, 0~0.000025% of a preservative and the balance being purified water.

[0025] The content and types of the moisturizing lubricant, buffer, and osmotic pressure regulator are the same as those described above, and will not be repeated here.

[0026] In one embodiment of the present invention, the mass content of the ion chelating agent can be 0.025~0.05%, or 0.05~0.1%; the ion chelating agent can be disodium ethylenediaminetetraacetate and / or sodium citrate. The ion chelating agent is used to complex calcium and magnesium ions in the system, inhibiting calcium phosphate mineral precipitation and lens protein calcification. The ion chelating agent is an excipient; high concentrations can cause slight irritation to damaged corneas, so it is generally not added to eye drops, eye rinses, or eye wash solutions.

[0027] In one embodiment of the present invention, the preservative content can be 0%, 0.00001~0.000015%, or 0.000015~0.000025% by mass; when the preservative content is not 0%, the mass ratio of the preservative to punicalin can be 1:(20~100); the preservative can be one or more of polyhexamethylene biguanide, sorbic acid, polyquaternium salt, and polyaminopropyl biguanide. The addition of preservatives at the above-mentioned mass contents can reduce toxicity and irritation to ocular surface cells, and simultaneously, through the combined action of punicalin and the preservative, maintain good antibacterial activity in eyeglass preservation solutions and contact lens care solutions. Even small amounts of the above-mentioned preservatives can cause slight irritation to damaged corneas, so they are generally not added to eye drops, eye rinses, and eye wash solutions.

[0028] The ophthalmic liquid composition provided by the present invention can achieve good antibacterial, antioxidant and anti-inflammatory activities by controlling each component and its content, while reducing toxicity and irritation to ocular surface cells and maintaining high ocular surface safety.

[0029] The present invention also provides a method for preparing the ophthalmic liquid composition described above, comprising: mixing punicin and a portion of purified water under light-protected conditions, then adding a moisturizing lubricant, a buffer, an osmotic pressure regulator and the remaining purified water for a second mixing, followed by filtration to obtain the ophthalmic liquid composition; Alternatively, it may include: under light-protected conditions, first mixing pungent glycoside and a portion of purified water, then adding a moisturizing lubricant, a buffer, an osmotic pressure regulator, an ion chelating agent, a preservative, and the remaining purified water for a second mixing, followed by filtration to obtain an ophthalmic liquid composition.

[0030] In this invention, pungent glycoside and a portion of purified water are first mixed under light-protected conditions.

[0031] Punic glycoside powder is easily decomposed by light, so the first mixing should be carried out under light-protected conditions. It is relatively stable after dissolving in water.

[0032] The present invention does not have any particular limitation on the method of the first mixing, as long as it can completely dissolve pungent glycoside in purified water.

[0033] After the first mixing, the present invention adds a moisturizing lubricant, a buffer, an osmotic pressure regulator and the remaining purified water for a second mixing and then filters, or adds a moisturizing lubricant, a buffer, an osmotic pressure regulator, an ion chelating agent, a preservative and the remaining purified water for a second mixing and then filters, to obtain an ophthalmic liquid composition.

[0034] As one embodiment of the present invention, the moisturizing lubricant, buffer, and osmotic pressure regulator may be added first, or the moisturizing lubricant, buffer, osmotic pressure regulator, ion chelating agent, and preservative may be added first, and then the container may be washed with the remaining purified water before the remaining purified water is added.

[0035] The present invention does not have any special limitations on the second mixing method, as long as it can completely dissolve and mix evenly.

[0036] In one embodiment of the present invention, the filter can be a PVDF (polyvinylidene fluoride) membrane with a pore size of 0.22 μm. This filtration can trap bacteria, mold, and spores introduced into the liquid during the mixing process; it can also remove insoluble impurities and tiny particles, ensuring uniform clarity.

[0037] The preparation method provided by this invention can obtain a sterile, uniform, and clear ophthalmic liquid composition.

[0038] This invention also provides the application of the ophthalmic liquid composition described in the above-described technical solutions or the ophthalmic liquid composition prepared by the preparation method described in the above-described technical solutions in eye drops, eye rinsing solutions, and eye wash solutions. In this invention, the ophthalmic liquid composition, when free of ion chelating agents and preservatives, is used in eye drops, eye rinsing solutions, and eye wash solutions.

[0039] This invention does not impose any particular limitation on the application described herein; any application method well-known to those skilled in the art can be used. When applied directly to the eyes as an eye wash, eye rinse, or eye cleanser, it can effectively reduce H2O2-induced oxidative damage to corneal epithelial cells and has antibacterial and anti-inflammatory effects against bacterial keratitis.

[0040] This invention also provides the application of the ophthalmic liquid composition described in the above-described technical solutions or the ophthalmic liquid composition prepared by the preparation method described in the above-described technical solutions in eyeglass preservation solutions and contact lens care solutions. In this invention, when the ophthalmic liquid composition contains an ion chelating agent (and a preservative), it is used in eyeglass preservation solutions and contact lens care solutions.

[0041] This invention does not impose any particular limitation on the application described herein; any application method well-known to those skilled in the art can be used. When applied to eyeglass preservation solutions and contact lens care solutions, it exhibits no cytotoxicity, excellent bactericidal effect, and good compatibility with lenses. The adsorption of pungent glycosides on lenses exhibits a "rapid adsorption-slow equilibrium" pattern, while the release exhibits a "burst release in the initial stage followed by a sustained release in the later stage." It can be slowly released during wear, providing continuous antibacterial, anti-inflammatory, and antioxidant protection to the ocular surface.

[0042] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0043] Example 1 Ophthalmic liquid composition: comprising 0.0005% pungent glycoside, 0.1% moisturizing lubricant (propylene glycol), 0.34% buffer (0.3% boric acid and 0.04% borax), 0.25% osmotic pressure regulator (sodium chloride), 0.025% ion chelating agent (disodium ethylenediaminetetraacetate), 0.000025% preservative (polyhexamethylene biguanide), and the balance purified water; Preparation method of ophthalmic liquid composition: Under light-protected conditions, pungent glycoside and a portion of purified water (accounting for 80% of the total purified water) are mixed to completely dissolve the pungent glycoside. Sodium chloride, boric acid, borax, disodium EDTA, propylene glycol, and polyhexamethylene biguanide are added and stirred until completely dissolved. The container is washed with the remaining purified water, and then the remaining purified water is added. The mixture is filtered using a PVDF (polyvinylidene fluoride) membrane with a pore size of 0.22 μm.

[0044] Example 2 The ophthalmic liquid composition differs from Example 1 only in that the amount of preservative (polyhexamethylene biguanide) is replaced with "0.000025%" instead of "0.000015%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0045] Example 3 The ophthalmic liquid composition differs from Example 1 only in that the amount of punicin is replaced by "0.0005%" to "0.001%" and the amount of preservative (polyhexamethylene biguanide) is replaced by "0.000025%" to "0.00001%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0046] Example 4 The ophthalmic liquid composition differs from Example 1 only in that the amount of punicin is replaced with "0.0005%" to "0.002%" and the preservative (polyhexamethylene biguanide) is removed; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0047] Example 5 Ophthalmic liquid composition: consisting of 0.0001% pungent glycoside, 0.005% moisturizing lubricant (sodium hyaluronate), 0.1422% buffer (0.1188% anhydrous disodium hydrogen phosphate and 0.0234% sodium dihydrogen phosphate monohydrate), 0.415% osmotic pressure regulator (sodium chloride), and the balance being purified water; Preparation method of ophthalmic liquid composition: Under light-protected conditions, pungent glycoside and a portion of purified water (accounting for 80% of the total purified water) are mixed to completely dissolve the pungent glycoside. Sodium chloride, anhydrous disodium hydrogen phosphate, sodium dihydrogen phosphate monohydrate, and sodium hyaluronate are added and stirred until completely dissolved. The container is washed with the remaining purified water, and then the remaining purified water is added. The mixture is filtered using a PVDF (polyvinylidene fluoride) membrane with a pore size of 0.22 μm.

[0048] Example 6 The ophthalmic liquid composition differs from that of Example 5 only in that the amount of pungent glycoside is replaced with "0.0003%" instead of "0.0001%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0049] Example 7 The ophthalmic liquid composition differs from that of Example 5 only in that the amount of pungent glycoside is replaced with "0.0005%" instead of "0.0001%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0050] Example 8 The ophthalmic liquid composition differs from that of Example 5 only in that the amount of pungent glycoside is replaced with "0.001%" instead of "0.001%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0051] Comparative Example 1 The ophthalmic liquid composition differs from Example 1 only in that pungent glycoside is removed and the amount of preservative (polyhexamethylene biguanide) is replaced with "0.04%" instead of "0.000025%"; The preparation method of the ophthalmic liquid composition is the same as in Example 1.

[0052] The ophthalmic liquid compositions prepared in Examples 1-8 and Comparative Example 1 were subjected to osmotic pressure testing using an instrument (Shanghai Medical University FM-8P freezing point osmoremeter); the ophthalmic liquid compositions prepared in Examples 1-8 and Comparative Example 1 were subjected to pH testing using an instrument (Mettler Toledo pH meter); the ophthalmic liquid compositions prepared in Examples 1-8 and Comparative Example 1 were subjected to cytotoxicity testing, specifically by culturing L929 mouse fibroblasts in a 37°C, 5% CO2 incubator until the logarithmic growth phase, and then adding 1.0 × 10⁻⁶ cells. 5 Cells were seeded at a concentration of [number] cells / mL into 96-well plates, with 100 μL of cell culture added to each well. The plates were incubated at 37°C with 5% CO2 for 24 hours. After complete cell adhesion, the original culture medium was discarded, and 100 μL of the culture medium for the ophthalmic liquid composition was added. Dimethyl sulfone (DMSO) was used as a positive control, and commercially available contact lens solution (Haichang Hydrating-SPA 60 mL) was used as a negative control. The plates were incubated for another 24 hours. After incubation, the supernatant was discarded, and 20 μL of thiazolyl blue (MTT) was added to each well. The plates were incubated for another 4 hours. After incubation, the supernatant was discarded, and 100 μL of LDMSO was added to each well. The absorbance (OD) of each well was measured using a microplate reader at a wavelength of 490-630 nm. Cell proliferation rate (RGR) = OD(sample) / OD(blank).

[0053] The physicochemical parameters (osmolarity, pH) and cytotoxicity test results of the obtained ophthalmic liquid composition are shown in Table 1.

[0054] Table 1. Physicochemical parameters (osmolarity, pH) and cytotoxicity test results of ophthalmic liquid compositions.

[0055] As can be seen from Table 1, the osmotic pressure of the ophthalmic liquid composition of the present invention is between 290 and 313 mOsm / kg, and the pH is between 7.20 and 7.24, which meets the requirements of YY0719.2-2022 standard. The RGR value of the ophthalmic liquid composition of the present invention is 85.66% to 91.59%, and it is non-cytotoxic (according to the requirements of YY0719.7-2011 standard, if the RGR value is greater than 80%, it indicates that the sample is non-cytotoxic).

[0056] Bactericidal experiments were conducted on the ophthalmic liquid compositions prepared in Examples 1-4 and Comparative Example 1. The bacteria used included Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli; the fungi used included Candida albicans and Aspergillus niger. All test bacteria were cultured on agar slant medium. Each culture was collected with sterile PBS, washed by centrifugation, and the optical density of the suspension was measured spectrophotometrically to estimate the approximate concentration of each bacterial culture. 10 mL of the ophthalmic liquid composition sample was added to each sample tube, and the bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli) and fungi (Candida albicans and Aspergillus niger) were inoculated into the sample tubes separately to achieve a bacterial concentration of 1.0 × 10⁻⁶. 5 CFU / mL ~1.0×10 6 Between CFU / mL, inoculate 3 tubes for each sample and each bacterial species, ensuring the inoculated volume does not exceed 1% of the sample volume. Mix thoroughly to ensure complete dispersion of the inoculated bacteria. Place the sample tubes in an incubator for cultivation. The incubation temperature is 35℃ for bacteria and 25℃ for fungi. On day 14, take 1.0 mL from each tube to determine the viable bacterial count. Take the average value of data from the same group. After sampling on day 14, re-inoculate each sample with bacterial suspension to achieve a bacterial concentration of 1.0 × 10⁻⁶. 4 CFU / mL ~ 1.0 × 10 5 CFU / mL, on day 28, take 1.0 mL from each tube to determine the viable bacterial count. According to the YY0719.4-2009 standard, for bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli), the bactericidal rate of each test bacterium should be no less than 99.9% on day 14; after re-inoculation on day 14, the concentration of each test bacterium should be further reduced on day 28, with a bactericidal rate no less than 99.9%. For fungi (Candida albicans and Aspergillus niger), on day 14, the number of recovered bacteria per milliliter for each test bacterium should remain at or below its initial concentration; after re-inoculation on day 14, the concentration of each test bacterium should remain at or below the concentration of each test bacterium after re-inoculation on day 28.

[0057] The bactericidal test results of the obtained ophthalmic liquid composition are shown in Table 2.

[0058] Table 2. Results of the bactericidal test of ophthalmic liquid compositions.

[0059] As can be seen from Table 2, the ophthalmic liquid composition of this invention showed no growth of common harmful microorganisms after 14 and 28 days of use after opening, which meets the requirements of YY0719.4-2009 standard; and the sterilization rate is comparable to, or even better than, Comparative Example 1 which only added chemical preservatives.

[0060] Lens compatibility tests were conducted on the ophthalmic liquid composition prepared in Example 1. Three experiments were performed: lens coloring, lens parameter influence, and adsorption and release. The lenses used in all three experiments were Haichang O2Balance Clear Oxygen Series silicone hydrogel contact lenses (material: PDMS1 / 2, water content 47%).

[0061] Lens staining experiment: Lenses were soaked in the ophthalmic liquid composition prepared in Example 1 for 7 days. The initial state of the lenses is shown in the photographs below. Figure 1 As shown in the photo, the condition of the lens after 7 days of soaking is as follows. Figure 2 As shown, the lens is uniformly stained with the natural polyphenol compound punicin, which can be observed with the naked eye.

[0062] Experiment on the influence of lens parameters: After soaking the lenses in the ophthalmic liquid composition prepared in Example 1 for a certain period of time (7 days, 14 days and 30 days), (the blank control group was "9% physiological saline" instead of "ophthalmic liquid composition"), the water content, oxygen permeability, base curve radius and diameter parameters of the lenses were tested. The obtained lens parameters are shown in Table 3.

[0063] Table 3. Lens parameters after soaking in ophthalmic liquid composition.

[0064] As can be seen from Table 3, the adsorption and release of pungent glycosides in the ophthalmic liquid composition have no adverse effects on key parameters such as the water content, oxygen permeability, diameter and light transmittance of the lens, indicating that the ophthalmic liquid composition of the present invention has good physical compatibility with contact lenses.

[0065] Adsorption and release experiments: The lens was immersed in a pungent glycoside-containing care solution, and the amount of pungent glycoside adsorbed by the lens was measured to obtain an adsorption kinetic curve; then the lens was placed in artificial tears, and the amount released was measured to obtain a release curve.

[0066] The specific method for determining the adsorption kinetic curve is as follows: The lens was placed in the ophthalmic liquid composition prepared in Example 1 of this invention, containing 0.0005% punicalin, and incubated at 37°C with constant temperature shaking. Samples were taken at 0.5, 1, 2, 4, 6, 8, 12, 24, and 48 hours, and the residual concentration of punicalin in the solution was determined by high performance liquid chromatography (HPLC). The adsorption amount of punicalin by the lens was calculated. Chromatographic conditions: C18 column (4.6 × 250 mm, 5 μm), mobile phase methanol-0.1% phosphoric acid aqueous solution (40:60), flow rate 1.0 mL / min, detection wavelength 378 nm. The obtained adsorption kinetic curve is shown below. Figure 3 As shown in Table 4, the adsorption kinetics data of pungent glycoside on silicone hydrogel lenses are presented.

[0067] Table 4 Adsorption kinetics data of pungent glycosides on silicone hydrogel lenses

[0068] From Table 4 and Figure 3 It can be seen that the adsorption of pungent glycosides on silicone hydrogel lenses exhibits a "rapid adsorption-slow equilibrium" characteristic; the adsorption rate is fastest within the first 2 hours, reaching 61.7% of the equilibrium adsorption capacity; after 8 hours, the adsorption tends to slow down; and adsorption equilibrium is reached at 48 hours, with an equilibrium adsorption capacity of 12.8 ± 0.1 μg / lens. The adsorption kinetic curve conforms to the pseudo-second-order kinetic model (R0). 2 =0.998), indicating that the adsorption process is dominated by chemisorption (hydrogen bonding, electrostatic interaction).

[0069] The specific method for determining the release curve is as follows: After adsorption equilibrium, the lens is removed, the surface is quickly rinsed with PBS, and transferred to a 24-well plate containing 2 mL of artificial tears. The plate is then incubated at 37°C with constant temperature shaking. Samples are taken at 0.5, 1, 2, 4, 6, 8, 12, and 24 hours, and an equal volume of fresh artificial tears is added. The concentration of punicin in the released solution is determined by HPLC. The release amount, cumulative release amount, and release rate at different time points are calculated, and the resulting release curve is shown below. Figure 4 As shown in Table 5, the release data of pungent glycosides on the silicone hydrogel lens are presented.

[0070] Table 5. Release data of pungent glycosides on silicone hydrogel lenses.

[0071] from Figure 4As shown in Table 5, the release of punicalin from the lens into artificial tears exhibits a two-stage characteristic: an initial burst release followed by a sustained release. The cumulative release rate reached 35.9±2.3% within the first hour, mainly due to the rapid desorption of punicalin physically adsorbed on the lens surface. Subsequently, a slow release phase ensued, with the cumulative release rate reaching 81.3±3.9% at 8 hours, 90.6±4.7% at 12 hours, and 96.9±4.7% at 24 hours. Kinetic model fitting of the release curves showed that the release curves conformed to the Higuchi kinetic model (R0). 2 =0.991), the release rate is linearly related to the square root of time, proving that the release of pungent glycoside from the lens into the tear film is mainly through diffusion mechanism. This sustained-release property can provide long-lasting antibacterial, anti-inflammatory and antioxidant protection for the ocular surface during wear.

[0072] The antioxidant properties of the ophthalmic liquid compositions prepared in Examples 1, 3-4 were tested. The specific testing method was as follows: human corneal epithelial cells in the logarithmic growth phase were taken and analyzed at a concentration of 1 × 10⁻⁶ cells per well. 4 Cells were seeded in 96-well plates and cultured for 24 hours. A blank control group, an H2O2 model control group (H2O2 concentration of 300 μmol / L), an example group, and a positive control group (vitamin C 100 μmol / L) were established. After pretreatment with the ophthalmic liquid composition prepared in Examples 1, 3, and 4 for 12 hours, 2 mL of 300 μmol / L H2O2 was added, and the cells were cultured for another 24 hours. The blank control group, H2O2 model control group, and positive control group were prepared by replacing the ophthalmic liquid composition with 9% physiological saline, 300 μmol / L H2O2, and 100 μmol / L vitamin C. 10 μL of LCK-8 reagent was added to each well, and after incubation for 2 hours, the OD value at 450 nm was measured using a microplate reader, and cell viability was calculated. The cell viability results after H2O2 oxidative damage are shown in Table 6.

[0073] Table 6. Cell viability results after H2O2 oxidative damage.

[0074] As shown in Table 6, compared with the model group, the cell survival rates of Examples 1, 3, and 4 of this invention increased in a concentration-dependent manner, reaching 68.7%, 74.2%, and 80.6%, respectively. Among them, the 0.002% punicalin group showed a 28.3% increase in cell survival rate compared with the model control. These results indicate that punicalin can effectively alleviate H2O2-induced oxidative damage to corneal epithelial cells and improve cell survival rate.

[0075] Anti-inflammatory experiments were conducted on the ophthalmic liquid compositions prepared in Examples 6-8. Specifically, a Pseudomonas aeruginosa (ATCC 19660) keratitis model was established using 6-8 week old female Swiss albino mice. Mice were divided into a model control group (no treatment), an example group, a commercially available control group (commercially available antibacterial eye drops), and a normal blank control group. Drug administration began 1 hour after modeling, four times daily for three consecutive days. Clinical scores were assessed on days 1, 2, and 3 post-infection. The criteria for clinical scoring are shown in Table 7, and the effect of different time points before and after treatment on bacterial keratitis in mice is shown in Table 8.

[0076] Table 7 Clinical Scoring Criteria

[0077] Table 8. Effects of treatment on mice with bacterial keratitis at different time points before and after treatment (scoring table)

[0078] As can be seen from Tables 7 and 8, compared with the model control group, the clinical scores of Examples 6-8 were significantly reduced, the corneal bacterial load was reduced, and the level of pro-inflammatory factors decreased in a concentration-dependent manner, demonstrating dual antibacterial and anti-inflammatory effects on bacterial keratitis.

[0079] As can be seen from the above embodiments, the ophthalmic liquid composition provided by the present invention is non-cytotoxic, has excellent bactericidal effect, good compatibility with lenses, exhibits "rapid adsorption-slow equilibrium" adsorption of pungent glycosides and "initial burst release-later sustained release" characteristics, can effectively reduce H2O2-induced oxidative damage to corneal epithelial cells and has antibacterial and anti-inflammatory effects on bacterial keratitis.

[0080] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An ophthalmic liquid composition, characterized in that, It includes punicein with a mass content of 0.0001~0.002%.

2. The ophthalmic liquid composition according to claim 1, characterized in that, It contains 0.0005 to 0.001% pungent glycosides by mass content.

3. The ophthalmic liquid composition according to claim 1 or 2, characterized in that, By mass content, it also includes 0.005-0.1% moisturizing lubricant, 0.01-0.5% buffer, 0.1-0.5% osmotic pressure regulator and the balance purified water.

4. The ophthalmic liquid composition according to claim 1 or 2, characterized in that, By mass content, it also includes 0.005~0.1% moisturizing lubricant, 0.01~0.5% buffer, 0.1~0.5% osmotic pressure regulator, 0.025~0.1% ion chelating agent, 0~0.000025% preservative and the balance purified water.

5. The ophthalmic liquid composition according to claim 4, characterized in that, When the content of the preservative is not 0, the mass ratio of the preservative to pungent glycoside is 1:(20~100).

6. The ophthalmic liquid composition according to claim 4, characterized in that, The preservative is one or more of polyhexamethylene biguanide, sorbic acid, polyquaternium salt, and polyaminopropyl biguanide.

7. The ophthalmic liquid composition according to claim 4, characterized in that, The ion chelating agent is disodium ethylenediaminetetraacetate and / or sodium citrate.

8. A method for preparing the ophthalmic liquid composition according to any one of claims 1 to 7, comprising: Under light-protected conditions, punicin and a portion of purified water were first mixed, then moisturizing lubricant, buffer, osmotic pressure regulator and the remaining purified water were added for a second mixing and then filtered to obtain an ophthalmic liquid composition. Alternatively, it may include: under light-protected conditions, first mixing pungent glycoside and a portion of purified water, then adding a moisturizing lubricant, a buffer, an osmotic pressure regulator, an ion chelating agent, a preservative, and the remaining purified water for a second mixing, followed by filtration to obtain an ophthalmic liquid composition.

9. The use of the ophthalmic liquid composition of claim 3 or the ophthalmic liquid composition prepared by the preparation method of claim 8 in eye drops, eye rinses and eye washes.

10. The use of the ophthalmic liquid composition according to any one of claims 4 to 7 or the ophthalmic liquid composition prepared by the preparation method according to claim 8 in eyeglass preservation solutions and contact lens care solutions.

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