Contact lens lubricating liquid containing lysozyme and recombinant collagen as well as preparation method and application of contact lens lubricating liquid

Abstract

The invention provides contact lens lubricating liquid containing lysozyme and recombinant collagen as well as a preparation method and application of the contact lens lubricating liquid, and belongs to the technical field of eye care products. The contact lens lubricating liquid provided by the invention comprises the following components: 0.01%-0.5% of lysozyme, 0.1%-2.0% of recombinant collagen, 0.5%-1.2% of an osmotic pressure regulator, 0.01%-0.1% of a viscosity regulator and 0.1%-2.0% of a pH buffer agent. The lysozyme and the recombinant collagen are taken as core active components, and the lysozyme and the recombinant collagen have a synergistic effect, so that the gel has the functions of resisting bacteria, preserving moisture, repairing cornea and improving wearing comfort. The invention further provides a preparation method of the contact lens lubricating liquid and application of the contact lens lubricating liquid in preparation of products for relieving xerophthalmia and preventing contact lens related keratitis.

Description

Technical Field

[0001] The invention belongs to the technical field of eye care products, and particularly relates to a contact lens lubricating liquid containing lysozyme and recombinant collagen, and a preparation method and application thereof. Background Art

[0002] Contact lenses play an important role in improving visual health due to their optical correction advantages. They are comfortable and convenient, making them popular among myopic patients, and the number of people wearing them has continued to grow in recent years. However, dry eyes are a common problem during contact lens wear, which not only leads to accumulation of deposits on the lens surface, decreased vision, reduced lens oxygen permeability, and changes in lens specifications, seriously affecting the quality of lens wear, but also significantly reducing the comfort of wearing the lenses. Consequently, contact lens lubricants have emerged, which are mainly used to alleviate the dryness and friction felt when wearing contact lenses and to keep the lens surface moist. However, contact lens lubricants currently on the market mostly rely on moisturizers such as hyaluronic acid and hydroxypropyl methylcellulose. Although they can alleviate dry eyes to a certain extent, most of them only have a simple lubricating and moisturizing effect, with relatively simple functions and limited effects on improving the ocular microenvironment and promoting ocular tissue repair. Furthermore, they usually need to rely on preservatives (such as benzalkonium chloride, etc.) to maintain sterility, which may increase the risk of the solution's biocompatibility not meeting requirements, and long-term use may cause eye irritation or allergic reactions.

[0003] Lysozyme is a natural antimicrobial protein widely used in medicine and the food industry. It can hydrolyze the peptidoglycan of bacterial cell walls, exerting a bactericidal effect. It is also highly safe and can replace chemical preservatives. Recombinant collagen is a new material developed using protein engineering and genetic recombination technologies. It has the advantages of being virus-free, having low rejection reactions, and being water-soluble. Although there are reports on the use of lysozyme or recombinant collagen in ophthalmic preparations, how to rationally combine these two ingredients and optimize the formula so that they can play a synergistic role in contact lens lubricants to produce a contact lens lubricant that can effectively inhibit microbial growth, promote corneal epithelial repair, and have good lubricating and moisturizing properties and low irritation has become an urgent problem to be solved in this field. Summary of the Invention

[0004] In order to solve the problems existing in the prior art, the purpose of the present invention is to provide a contact lens lubricant containing lysozyme and recombinant collagen and a preparation method thereof. Lysozyme and recombinant collagen are used as core active ingredients, and the two act synergistically to have the functions of antibacterial, moisturizing, corneal repair and improving wearing comfort.

[0005] The present invention also aims to provide the use of the above-mentioned contact lens lubricating liquid in the preparation of products for relieving dry eye and preventing contact lens-related keratitis.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a contact lens lubricating liquid containing lysozyme and recombinant collagen. The lubricating liquid comprises the following components, calculated by mass volume percentage: 0.01% to 0.5% of lysozyme, 0.1% to 2.0% of recombinant collagen, 0.5% to 1.2% of an osmotic pressure regulator, 0.01% to 0.1% of a viscosity regulator, and 0.1% to 2.0% of a pH buffer.

[0008] Preferably, the contact lens lubricating solution does not contain chemical preservatives.

[0009] Preferably, the osmotic pressure regulator comprises sodium chloride and / or glycerol.

[0010] Preferably, the viscosity modifier comprises hydroxypropyl methylcellulose and / or sodium carboxymethylcellulose.

[0011] Preferably, the pH buffer comprises a boric acid-borax buffer and / or a phosphate buffer.

[0012] The present invention also provides a method for preparing the above-mentioned contact lens lubricating liquid, comprising the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water, stirring until clarified, to obtain component A; adding lysozyme to component A, stirring and dissolving at low temperature, then adding recombinant collagen, heating and hydrating to obtain component B; adding a viscosity regulator to component B, mixing, and sterilizing to obtain the contact lens lubricating liquid.

[0013] Preferably, the temperature of the low-temperature stirring is 2-8°C.

[0014] Preferably, the temperature is raised to 25-30°C.

[0015] Preferably, the sterilization is sterilization by filtration using a 0.22 μm filter membrane.

[0016] The present invention also provides the use of the contact lens lubricating liquid in preparing products for relieving dry eye and preventing contact lens-related keratitis.

[0017] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0018] The contact lens lubricant provided by the present invention uses lysozyme and recombinant collagen as core ingredients. Lysozyme inhibits bacterial growth, and recombinant collagen forms a hydrophilic film on the lens surface, reducing friction and repairing micro-damage. The two act synergistically to have the functions of antibacterial, moisturizing, repairing the cornea, and improving wearing comfort. In addition, the contact lens lubricant has low irritation and a low risk of eye infection, making it suitable for sensitive people. DETAILED DESCRIPTION

[0019] The present invention provides a contact lens lubricating solution containing lysozyme and recombinant collagen, comprising the following components, calculated by mass volume percentage: 0.01% to 0.5% lysozyme, 0.1% to 2.0% recombinant collagen, 0.5% to 1.2% osmotic pressure regulator, 0.01% to 0.1% viscosity regulator, and 0.1% to 2.0% pH buffer. Preferably, the contact lens lubricating solution comprises the following components: 0.1% to 0.3% lysozyme, 0.5% to 1.2% recombinant collagen, 0.6% to 1.0% osmotic pressure regulator, 0.05% to 0.08% viscosity regulator, and 0.15% to 0.5% pH buffer. The mass-to-volume ratios described herein are expressed in g / mL. The contact lens lubricating solution also comprises deionized water, preferably comprising the remainder of the components.

[0020] The contact lens lubricant described herein does not contain chemical preservatives. The added lysozyme provides broad-spectrum antimicrobial activity, replacing traditional preservatives. The lubricant exhibits an antibacterial rate of >99.9% against Staphylococcus aureus. The recombinant collagen in the lubricant forms a hydrophilic film on the lens surface, promoting corneal repair and reducing damage caused by mechanical friction. It works synergistically with lysozyme to provide antibacterial, moisturizing, corneal repair, and improved wearing comfort. The lubricant's moisturizing properties are 40% longer than those of traditional hyaluronic acid formulations. Furthermore, the lubricant is non-irritating, with a Draize score of ≤1, making it suitable for sensitive individuals. The sources of the lysozyme and recombinant collagen are not limited; commercially available products can be used.

[0021] The osmotic pressure regulator of the present invention includes sodium chloride and / or glycerol. By adding the osmotic pressure regulator, the present invention can adjust the contact lens lubricating solution to be isotonic with tears (280-320 mOsm / kg); the viscosity regulator of the present invention includes hydroxypropyl methylcellulose and / or sodium carboxymethyl cellulose. By adding the viscosity regulator, the present invention prolongs the wetting time; the pH buffer of the present invention includes boric acid-borax buffer and / or phosphate buffer. By adding the pH buffer, the present invention adjusts the pH of the contact lens lubricating solution to 6.5-7.5.

[0022] The present invention also provides a method for preparing the above-mentioned contact lens lubricating liquid, comprising the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water, stirring until clarified, to obtain component A; adding lysozyme to component A, stirring and dissolving at low temperature, then adding recombinant collagen, heating and hydrating to obtain component B; adding a viscosity regulator to component B, mixing, and sterilizing to obtain the contact lens lubricating liquid.

[0023] The temperature of low-temperature stirring of the present invention is 2-8°C, which can be optionally 3°C, 4°C, 5°C, 6°C or 7°C. The present invention dissolves lysozyme by low-temperature stirring, thereby avoiding the inactivation of lysozyme; the heating described in the present application is to be heated to 25-30°C, which can be optionally 26°C, 27°C, 28°C or 29°C, that is, after adding recombinant collagen, the temperature is slowly increased to fully hydrate it. The hydration described in the present invention refers to hydration (also known as hydration); the slow heating is preferably heated at 0.1-0.5°C / min. The present invention ensures the activity of lysozyme and collagen by controlling pH and low-temperature preparation process to ensure that both play antibacterial, moisturizing, corneal repair and other effects.

[0024] The sterilization of the present invention is preferably sterilization by filtration with a 0.22 μm filter membrane. The present invention can optionally include aseptically packaging the contact lens lubricating liquid after filtration and sterilization to obtain the product.

[0025] The present invention also provides the use of the above-mentioned contact lens lubricating liquid in preparing a product for relieving dry eye and preventing contact lens-related keratitis. The product preferably contains 50 to 100 wt% of the contact lens lubricating liquid of the present invention.

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0027] In the specific embodiment of the present invention, lysozyme (Sigma), production batch number WXBF2947V; recombinant collagen was purchased from Jiangsu Chuangshi Medical Technology Co., Ltd., production batch number F2410003.

[0028] In the following examples, unless otherwise specified, all methods are conventional.

[0029] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0030] Example 1

[0031] A contact lens lubricant containing lysozyme and recombinant collagen comprises the following components, calculated by mass volume (g / mL): 0.01% lysozyme, 0.1% recombinant collagen, 0.5% osmotic pressure regulator (glycerol), 0.01% viscosity regulator (sodium carboxymethyl cellulose), and 0.1% pH buffer (boric acid-borax buffer, pH 7.0).

[0032] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clear to obtain component A; adding lysozyme to component A and stirring to dissolve the mixture at 2°C; then adding recombinant collagen and heating the mixture to 25°C at a rate of 0.1°C / min to fully hydrate the mixture to obtain component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0033] Example 2

[0034] A contact lens lubricant containing lysozyme and recombinant collagen comprises the following components, calculated by mass volume (g / mL): 0.5% lysozyme, 2.0% recombinant collagen, 1.2% osmotic pressure regulator (sodium chloride), 0.1% viscosity regulator (hydroxypropyl methylcellulose), and 2% pH buffer (phosphate buffer, pH 6.8).

[0035] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clear to obtain component A; adding lysozyme to component A and stirring and dissolving the mixture at 8°C; then adding recombinant collagen and heating the mixture to 30°C at a rate of 0.5°C / min to fully hydrate the mixture to obtain component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0036] Example 3

[0037] A contact lens lubricant containing lysozyme and recombinant collagen comprises the following components, calculated by mass volume (g / mL): 0.2% lysozyme, 0.8% recombinant collagen, 0.9% osmotic pressure regulator (sodium chloride), 0.05% viscosity regulator (sodium carboxymethyl cellulose), and 0.8% pH buffer (boric acid-borax buffer, pH 7.0).

[0038] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clarified to obtain component A; adding lysozyme to component A and stirring to dissolve the mixture at 4°C; then adding recombinant collagen and heating the mixture to 28°C at a rate of 0.2°C / min to fully hydrate the mixture to obtain component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0039] Example 4

[0040] A contact lens lubricant containing lysozyme and recombinant collagen comprises the following components, calculated by mass volume (g / mL): 0.3% lysozyme, 1.0% recombinant collagen, 1.0% osmotic pressure regulator (glycerol), 0.03% viscosity regulator (hydroxypropyl methylcellulose), and 1.2% pH buffer (phosphate buffer, pH 6.8).

[0041] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clear to obtain component A; adding lysozyme to component A and stirring and dissolving the mixture at 4°C; then adding recombinant collagen and heating the mixture to 26°C at a rate of 0.3°C / min to fully hydrate the mixture to obtain component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0042] Comparative Example 1

[0043] Compared with Example 3, the addition of lysozyme and recombinant collagen was omitted (double negative control group):

[0044] Calculated by mass volume (g / mL), the composition includes the following components: 0.9% of an osmotic pressure regulator (sodium chloride), 0.05% of a viscosity regulator (sodium carboxymethyl cellulose), and 0.8% of a pH buffer (boric acid-borax buffer, pH 7.0).

[0045] Preparation method steps: dissolve the buffer and osmotic pressure regulator in deionized water, stir until clear, and obtain component A; add viscosity regulator to component A, mix, and then sterilize.

[0046] Comparative Example 2

[0047] Compared with Example 4, the addition of lysozyme and recombinant collagen was omitted (double negative control group):

[0048] Calculated by mass volume (g / mL), the composition includes the following components: 1.0% osmotic pressure regulator (glycerol), 0.03% viscosity regulator (hydroxypropyl methylcellulose), and 1.2% pH buffer (phosphate buffer, pH 6.8).

[0049] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until the solution is clear to obtain component A; adding a viscosity regulator to component A, mixing the mixture and sterilizing the mixture to obtain a contact lens lubricant.

[0050] Comparative Example 3

[0051] Compared with Example 3, the addition of lysozyme was omitted (lysozyme negative control):

[0052] Calculated by mass volume (g / mL), the composition includes the following components: 0.8% recombinant collagen, 0.9% osmotic pressure regulator (sodium chloride), 0.05% viscosity regulator (sodium carboxymethyl cellulose), and 0.8% pH buffer (boric acid-borax buffer, pH 7.0).

[0053] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clarified to obtain component A; adding recombinant collagen to component A, heating the mixture to 28°C at a rate of 0.2°C / min to fully hydrate the mixture, and obtaining component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0054] Comparative Example 4

[0055] Compared with Example 4, the addition of lysozyme was omitted (lysozyme negative control):

[0056] Calculated by mass volume (g / mL), the composition includes the following components: 1.0% recombinant collagen, 1.0% osmotic pressure regulator (glycerol), 0.03% viscosity regulator (hydroxypropyl methylcellulose), and 1.2% pH buffer (phosphate buffer, pH 6.8).

[0057] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clarified to obtain component A; adding recombinant collagen to component A, heating the mixture to 26°C at a rate of 0.3°C / min to fully hydrate the mixture, and obtaining component B; adding a viscosity regulator to component B, mixing the mixture, and sterilizing the mixture to obtain a contact lens lubricant.

[0058] Comparative Example 5

[0059] Compared with Example 3, the addition of recombinant collagen was omitted (recombinant collagen negative control):

[0060] Calculated by mass volume (g / mL), the composition includes the following components: 0.2% lysozyme, 0.9% osmotic pressure regulator (sodium chloride), 0.05% viscosity regulator (sodium carboxymethyl cellulose), and 0.8% pH buffer (boric acid-borax buffer, pH 7.0).

[0061] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clear to obtain component A; adding lysozyme to component A and stirring and dissolving the mixture at 4° C. to obtain component B; adding a viscosity regulator to component B, mixing the mixture and sterilizing the mixture to obtain a contact lens lubricant.

[0062] Comparative Example 6

[0063] Compared with Example 4, the addition of recombinant collagen was omitted (recombinant collagen negative control):

[0064] Calculated by mass volume (g / mL), the invention comprises the following components: 0.3% lysozyme, 1.0% osmotic pressure regulator (glycerol), 0.03% viscosity regulator (hydroxypropyl methylcellulose), and 1.2% pH buffer (phosphate buffer, pH 6.8).

[0065] The preparation method comprises the following steps: dissolving a buffer and an osmotic pressure regulator in deionized water and stirring until clear to obtain component A; adding lysozyme to component A and stirring and dissolving the mixture at 4° C. to obtain component B; adding a viscosity regulator to component B, mixing the mixture and sterilizing the mixture to obtain a contact lens lubricant.

[0066] Test Example 1

[0067] The following experiments were conducted using Examples 3 to 4 and Comparative Examples 1 to 6, and existing products were used as controls:

[0068] (1) Antibacterial test: The test method is in accordance with YY / T0719.3-20090719 (24 hours). The conventional lubricant containing biguanide preservatives is the brand of Shanghai Weikang Optical Glasses Co., Ltd., with the trade name Weikang Mei. The test results are as follows:

[0069] Table 1 Antibacterial test results

[0070]

[0071] The results showed that the contact lens lubricant containing lysozyme and recombinant collagen provided by the present invention had good antibacterial properties against Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC9027, Escherichia coli ATCC8739, Candida albicans ATCC10231, and Fusarium solani ATCC36031. The antibacterial effect was comparable to or slightly higher than that of commercially available lubricants containing biguanide preservatives. After omitting the addition of recombinant collagen, the antibacterial effect decreased, and after omitting the addition of lysozyme, the antibacterial effect decreased significantly. After omitting both recombinant collagen and lysozyme, the antibacterial effect was very poor.

[0072] (2) Lubricity test: The test method is in accordance with GB / T 10006-2021 (friction material: float glass; ground material: white silica gel). The commercially available lubricant containing sodium hyaluronate is the brand of Shanghai Weikang Optical Glasses Co., Ltd., and the product name is Weikang Mei. The test results are as follows:

[0073] Table 2 Lubricity test results

[0074]

[0075]

[0076] The results showed that the contact lens lubricant containing lysozyme and recombinant collagen provided by the present invention has good lubricity and is better than the commercially available lubricant containing sodium hyaluronate. After abandoning the addition of recombinant collagen, the lubrication effect decreased significantly, and after abandoning the addition of lysozyme, the lubrication effect also decreased. After abandoning both recombinant collagen and lysozyme, the lubrication effect was poor and close to that of normal saline.

[0077] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A contact lens lubricating solution containing lysozyme and recombinant collagen, characterized in that: Calculated by mass volume percentage, the composition includes: 0.01% to 0.5% of lysozyme, 0.1% to 2.0% of recombinant collagen, 0.5% to 1.2% of osmotic pressure regulator, 0.01% to 0.1% of viscosity regulator and 0.1% to 2.0% of pH buffer.

2. The contact lens lubricating liquid according to claim 1, wherein The contact lens lubricating solution does not contain chemical preservatives.

3. The contact lens lubricating liquid according to claim 1, wherein The osmotic pressure regulator includes sodium chloride and / or glycerol.

4. The contact lens lubricating liquid according to claim 1, wherein The viscosity modifier includes hydroxypropyl methylcellulose and / or sodium carboxymethylcellulose.

5. The contact lens lubricating liquid according to claim 1, wherein The pH buffer includes a boric acid-borax buffer and / or a phosphate buffer.

6. The method for preparing a contact lens lubricating liquid according to any one of claims 1 to 5, characterized in that: The following steps are involved: The buffer and osmotic pressure regulator are dissolved in deionized water and stirred until clear to obtain component A; lysozyme is added to component A and dissolved by stirring at low temperature, and then recombinant collagen is added and hydrated by heating to obtain component B; a viscosity regulator is added to component B, and the mixture is mixed and sterilized to obtain a contact lens lubricant.

7. The preparation method according to claim 6, characterized in that The temperature of the low-temperature stirring is 2-8°C.

8. The preparation method according to claim 6, characterized in that The heating is to 25-30°C.

9. The preparation method according to claim 6, characterized in that The sterilization is sterilization by filtration using a 0.22 μm filter membrane.

10. Use of the contact lens lubricating liquid according to any one of claims 1 to 5 in the preparation of a product for relieving dry eye and preventing contact lens-related keratitis.

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