A kind of protein eye protection liquid and preparation process thereof

Abstract

The present invention relates to the technical field of eye medical products, and in particular to a protein eye protection liquid and a preparation process thereof. The preparation process comprises the following steps: preparing albumin nanoparticles; preparing astragalus glycoside; preparing astragalus glycoside nano-albumin; and preparing astragalus glycoside nano-albumin eye drops. The present invention couples albumin nanoparticles with astragalus glycoside, and uses nanoparticles to coat astragalus glycoside, thereby reducing the irritation of astragalus glycoside. Moreover, the astragalus glycoside coated with nanoparticles can prevent astragalus glycoside from aggregating in the eye protection liquid, thereby ensuring the stability and dispersibility of the eye protection liquid, and is slowly released in the eye protection liquid, thereby ensuring the stability of the astragalus glycoside efficacy, and being more gentle, thereby achieving the effect of eye protection.

Description

Technical Field

[0001] The invention relates to the technical field of eye medical products, and in particular to a protein eye protection solution and a preparation process thereof. Background Art

[0002] Myopia is becoming more and more common among modern people. Some people wear contact lenses for beauty. After wearing contact lenses, their eyes will experience problems such as fatigue, dryness and bacterial invasion. Therefore, eye protection solutions that can relieve eye fatigue and protect eye health have also appeared.

[0003] Most of the existing eye protection solutions use ingredients that can soothe eye cells, such as collagen, to protect the eyes. However, they do little to solve the problem of bacterial invasion. In addition, the protein components in some eye protection solutions will provide nutrients for bacteria and have a low bactericidal effect. Some eye protection solutions containing bactericidal ingredients are irritating to the eyes because they are mainly used for bactericidal purposes. Therefore, they have become objects that people refuse to choose. Therefore, there is a need for an eye protection solution that has bactericidal ingredients and is low or non-irritating to the eyes, so as to solve the deficiencies of the existing technology. Summary of the invention

[0004] In view of the deficiencies in the prior art, the object of the present invention is to provide a protein eye protection solution and a preparation process thereof.

[0005] A preparation process of a protein eye protection solution comprises the following steps:

[0006] S1: Preparation of albumin nanoparticles

[0007] Weigh bovine serum albumin, dissolve it in a PBS solution, add a composite emulsifier to obtain a solution to be emulsified, add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification, and then perform rotary evaporation, then dropwise add anhydrous ethanol under stirring until a colloidal solution is formed, add glutaraldehyde to the colloidal solution, continue stirring and solidifying, wash after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain albumin nanoparticles;

[0008] S2: Preparation of Astragalus glycosides

[0009] The enzyme solution of kaempferol substrate, sucrose, UDP, Tris-HCl buffer, MgCl, DMSO, FCT and GmSUS is reacted at 32-35°C and 200-250 r / min for 1-4 hours, methanol is added to terminate the reaction, and the supernatant is obtained by centrifugation to obtain reaction solution I, and the reaction solution I is centrifuged twice, and the obtained supernatant is added to an LH-20 dextran gel column, equilibrated and eluted with an ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside;

[0010] S3: Preparation of Astragalus glycoside nanoalbumin

[0011] Dissolve astragalus glycoside in deionized water to obtain an astragalus glycoside aqueous solution, adjust the pH of the astragalus glycoside aqueous solution to 4-5.5 with dilute hydrochloric acid, add EDC and NHS in sequence, stir and activate in the dark, mix the albumin nanoparticles with astragalus glycoside, wash after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin;

[0012] S4: Preparation of Astragalus glycoside nanoalbumin eye drops

[0013] Hydroxypropyl-β-cyclodextrin and deionized water were magnetically stirred at a constant temperature until dissolved, and mannitol was added to obtain a saturated aqueous solution. Astragalus glycoside was weighed and slowly added to the saturated aqueous solution. After ultrasound, water was added to 100 parts by volume to obtain an intermediate solution, and astragalus glycoside nanoalbumin was added and stirred at room temperature overnight. After filtering with a 0.22 μm filter membrane, astragalus glycoside nanoalbumin eye drops were obtained.

[0014] Furthermore, step S1 of preparing albumin nanoparticles comprises the following steps:

[0015] S1.1: Weigh bovine serum albumin and dissolve it in PBS solution. The solid-liquid ratio of bovine serum albumin to PBS solution is 1:(18-20). The pH of the system is adjusted to 7.5-8. A composite emulsifier with a concentration of 0.5-0.75% is added to obtain a solution to be emulsified.

[0016] S1.2: Add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification for 10-15 minutes, then transfer to a rotary evaporator for rotary evaporation for 10-15 minutes, and then add anhydrous ethanol dropwise under stirring until a colloidal solution is formed;

[0017] S1.3: Add glutaraldehyde to the colloidal solution and continue stirring to solidify for 1-2 hours. After high-speed centrifugation, wash 1-3 times, collect the precipitate, and freeze-dry to obtain albumin nanoparticles.

[0018] Furthermore, step S2 of preparing astragaloside comprises the following steps:

[0019] S2.1: 8-10 parts by volume of 1-2 mmol / L kaempferol substrate, 100-110 mmol / L sucrose, 0.2-0.3 mmol / L UDP, 100-120 mmol / L Tris-HCl buffer, 50-60 mmol / LMgCl, 5-6% DMSO, FCT and GmSUS enzyme solution were reacted at 32-35°C and 200-250 r / min for 1-4 h, 20-30 parts by volume of methanol were added to terminate the reaction, and the mixture was centrifuged at 12000-15000 r / min for 8-10 min at 0-4°C to obtain the supernatant to obtain reaction solution I;

[0020] S2.2: The reaction solution I is centrifuged twice at 12000-15000 r / min for 10-15 min, and the obtained supernatant is added to an LH-20 polysaccharide gel column, equilibrated and eluted with 45-50% ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside.

[0021] Further, step S3 prepares astragalus glycoside nano-albumin, comprising the following steps:

[0022] S3.1: Dissolve 1-5 parts by volume of astragaloside in 10-15 parts by volume of deionized water to obtain an astragaloside aqueous solution, adjust the pH of the astragaloside aqueous solution to 4-5.5 with dilute hydrochloric acid, add EDC and NHS in a ratio of 1:(1.5-2):(1.2-1.5) in sequence, and stir to activate for 1-2 hours in a dark place;

[0023] S3.2: Mix albumin nanoparticles and astragalus glycoside in a mass ratio of 1:(4-5), adjust the pH of the system to 7-7.5 with PBS solution, stir the reaction for 12-14 hours, wash 1-3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin.

[0024] Further, step S4 prepares astragalus glycoside nano-albumin eye drops, comprising the following steps:

[0025] S4.1: Place 25-30 parts by mass of hydroxypropyl-β-cyclodextrin and 40-45 parts by volume of deionized water in an environment of 60-65°C with constant temperature magnetic stirring until dissolved, and then add 0.15-0.2 parts by mass of mannitol to obtain a saturated aqueous solution;

[0026] S4.2: Weigh 0.2-1 parts by mass of astragalus glycoside, slowly add it to a saturated aqueous solution at 60-65°C, sonicate for 60-65 minutes, and add water to 100 parts by volume to obtain an intermediate solution;

[0027] S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. Filter with 0.22 μm filter membrane to obtain astragalus glycoside nano-albumin eye drops.

[0028] Furthermore, in step S1, the composite emulsifier is prepared by mixing sodium caseinate and poloxamer in a volume ratio of 2:1.

[0029] Furthermore, the pH of the Tris-HCl buffer in step S2 is 7.

[0030] Furthermore, in step S1, the volume ratio of the colloidal solution to glutaraldehyde is 1:(0.2-0.5).

[0031] Furthermore, in step S4, the volume ratio of astragalus glycoside nano-albumin to the intermediate solution is 1:(2-3).

[0032] A protein eye protection liquid is prepared by the preparation process of the protein eye protection liquid.

[0033] Compared with the prior art, the present invention has at least the following beneficial effects:

[0034] 1. The present invention couples albumin nanoparticles with astragalus glycoside and uses the nanoparticles to coat astragalus glycoside, thereby reducing the irritation of astragalus glycoside. In addition, the astragalus glycoside coated with the nanoparticles can prevent the astragalus glycoside from aggregating in the eye protection solution, thereby ensuring the stability and dispersibility of the eye protection solution, and slowly releases the astragalus glycoside in the eye protection solution, thereby ensuring the stability of the efficacy of the astragalus glycoside and being more gentle, thereby achieving the effect of protecting the eyes.

[0035] 2. The present invention includes astragalus glycosides through β-cyclodextrin. Cyclodextrin as the outer coating of the inclusion complex is easy to dissociate and will not change the medicinal properties of the inclusion complex. In this way, the drug can enter the eyes with reduced irritation and enhanced stability. When coupled with astragalus glycosides by albumin nanoparticles, irritation can be reduced while ensuring the concentration of astragalus glycosides. The onset time of the eye protection solution can be prolonged by slow release, and the quality of the eye protection solution can be made more stable. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the present disclosure and, together with the description, further serve to explain the principles of the present disclosure and to enable those skilled in the relevant art to make and use the present disclosure.

[0037] Figure 1 The present invention is a flow chart of the preparation process of a protein eye protection solution used in an embodiment of the present invention. DETAILED DESCRIPTION

[0038] The following is a detailed description of a protein eye protection solution and its preparation process provided by the present invention in conjunction with the accompanying drawings and specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.

[0039] Embodiment 1:

[0040] A preparation process of protein eye protection solution, such as Figure 1 As shown, the following steps are included:

[0041] S1: Preparation of albumin nanoparticles

[0042] S1.1 Weigh bovine serum albumin and dissolve it in PBS solution. The solid-liquid ratio of bovine serum albumin to PBS solution is 1:20. The pH of the system is adjusted to 7.5. A composite emulsifier with a concentration of 0.75% is added. The composite emulsifier is a mixture of sodium caseinate and poloxamer in a volume ratio of 2:1 to obtain an emulsified solution.

[0043] S1.2: Add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification for 15 minutes, then transfer to a rotary evaporator for rotary evaporation for 10 minutes, and then add anhydrous ethanol dropwise under stirring until a colloidal solution is formed;

[0044] S1.3: Add glutaraldehyde to the colloidal solution and continue stirring to solidify for 1 hour. The volume ratio of the colloidal solution to glutaraldehyde is 1:0.2. After high-speed centrifugation, wash three times, collect the precipitate, and freeze-dry to obtain albumin nanoparticles.

[0045] S2: Preparation of Astragalus glycosides

[0046] S2.1: 10 parts by volume of 1 mmol / L kaempferol substrate, 100 mmol / L sucrose, 0.2 mmol / L UDP, 120 mmol / L Tris-HCl buffer (pH = 7), 60 mmol / LMgCl, 5% DMSO, FCT and GmSUS enzyme solution were reacted at 35°C and 200 r / min for 4 h, 30 parts by volume of methanol were added to terminate the reaction, and the mixture was centrifuged at 15000 r / min for 10 min at 4°C to obtain the supernatant to obtain reaction solution I;

[0047] S2.2: The reaction solution I is centrifuged twice at 15000 r / min for 15 min, and the obtained supernatant is added to an LH-20 polysaccharide gel column, equilibrated and eluted with a 50% ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside.

[0048] S3: Preparation of Astragalus glycoside nanoalbumin

[0049] S3.1: Dissolve 1 part by volume of astragaloside in 10 parts by volume of deionized water to obtain an astragaloside aqueous solution, adjust the pH of the astragaloside aqueous solution to 5.5 with dilute hydrochloric acid, add EDC and NHS in a ratio of 1:2:1.2, and activate with stirring for 2 hours in a dark place;

[0050] S3.2: Mix albumin nanoparticles and astragalus glycoside in a mass ratio of 1:5, adjust the pH of the system to 7.5 with PBS solution, stir the reaction for 12 hours, wash 3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin.

[0051] S4: Preparation of Astragalus glycoside nanoalbumin eye drops

[0052] S4.1: 25 parts by mass of hydroxypropyl-β-cyclodextrin and 45 parts by volume of deionized water are placed in an environment of 65°C and magnetically stirred until dissolved, and then 0.2 parts by mass of mannitol are added to obtain a saturated aqueous solution;

[0053] S4.2: Weigh 1 part by mass of astragalus glycoside, slowly add it into a saturated aqueous solution at 65°C, sonicate for 60 minutes, and add water to 100 parts by volume to obtain an intermediate solution;

[0054] S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. The volume ratio of astragalus glycoside nano-albumin and the intermediate solution is 1:2. After filtering with a 0.22 μm filter membrane, astragalus glycoside nano-albumin eye drops are obtained.

[0055] Embodiment 2:

[0056] A preparation process of protein eye protection solution, such as Figure 1 As shown, the following steps are included:

[0057] S1: Preparation of albumin nanoparticles

[0058] S1.1 Weigh bovine serum albumin and dissolve it in PBS solution. The solid-liquid ratio of bovine serum albumin to PBS solution is 1:18. The pH of the system is adjusted to 7.5. A composite emulsifier with a concentration of 0.75% is added. The composite emulsifier is a mixture of sodium caseinate and poloxamer in a volume ratio of 2:1 to obtain an emulsified solution.

[0059] S1.2: Add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification for 15 minutes, then transfer to a rotary evaporator for rotary evaporation for 10 minutes, and then add anhydrous ethanol dropwise under stirring until a colloidal solution is formed;

[0060] S1.3: Add glutaraldehyde to the colloidal solution and continue stirring to solidify for 1 hour. The volume ratio of the colloidal solution to glutaraldehyde is 1:0.2. After high-speed centrifugation, wash three times, collect the precipitate, and freeze-dry to obtain albumin nanoparticles.

[0061] S2: Preparation of Astragalus glycosides

[0062] S2.1: 8 volumes of each of 1mmol / L kaempferol substrate, 100mmol / L sucrose, 0.2mmol / L UDP, 120mmol / L Tris-HCl buffer (pH=7), 60mmol / LMgCl, 5% DMSO, FCT and GmSUS enzyme solution were reacted at 35℃, 200r / min for 4h, 20 volumes of methanol were added to terminate the reaction, and the mixture was centrifuged at 15000r / min for 10min at 4℃ to obtain the supernatant to obtain reaction solution I;

[0063] S2.2: The reaction solution I is centrifuged twice at 15000 r / min for 15 min, and the obtained supernatant is added to an LH-20 polysaccharide gel column, equilibrated and eluted with a 50% ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside.

[0064] S3: Preparation of Astragalus glycoside nanoalbumin

[0065] S3.1: Dissolve 5 parts by volume of astragalus glycoside in 15 parts by volume of deionized water to obtain an astragalus glycoside aqueous solution, adjust the pH of the astragalus glycoside aqueous solution to 5.5 with dilute hydrochloric acid, add EDC and NHS in a ratio of 1:1.5:1.5, and activate with stirring for 2 hours in a dark place;

[0066] S3.2: Mix albumin nanoparticles and astragalus glycoside in a mass ratio of 1:4, adjust the pH of the system to 7.5 with PBS solution, stir the reaction for 12 hours, wash 3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin.

[0067] S4: Preparation of Astragalus glycoside nanoalbumin eye drops

[0068] S4.1: 30 parts by mass of hydroxypropyl-β-cyclodextrin and 40 parts by volume of deionized water are placed in an environment of 65°C and magnetically stirred until dissolved, and then 0.2 parts by mass of mannitol are added to obtain a saturated aqueous solution;

[0069] S4.2: Weigh 0.2 parts by mass of astragalus glycoside, slowly add it into a saturated aqueous solution at 65°C, sonicate for 60 minutes, and add water to 100 parts by volume to obtain an intermediate solution;

[0070] S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. The volume ratio of astragalus glycoside nano-albumin and the intermediate solution is 1:2.5. Filter with a 0.22 μm filter membrane to obtain astragalus glycoside nano-albumin eye drops.

[0071] Embodiment 3:

[0072] A preparation process of protein eye protection solution, such as Figure 1 As shown, the following steps are included:

[0073] S1: Preparation of albumin nanoparticles

[0074] S1.1 Weigh bovine serum albumin and dissolve it in PBS solution. The solid-liquid ratio of bovine serum albumin to PBS solution is 1:20. The pH of the system is adjusted to 8. A composite emulsifier with a concentration of 0.75% is added. The composite emulsifier is a mixture of sodium caseinate and poloxamer in a volume ratio of 2:1 to obtain an emulsified solution.

[0075] S1.2: Add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification for 10 minutes, then transfer to a rotary evaporator for rotary evaporation for 10 minutes, and then add anhydrous ethanol dropwise under stirring until a colloidal solution is formed;

[0076] S1.3: Add glutaraldehyde to the colloidal solution and continue stirring to solidify for 2 hours. The volume ratio of the colloidal solution to glutaraldehyde is 1:0.2. After high-speed centrifugation, wash three times, collect the precipitate, and freeze-dry to obtain albumin nanoparticles.

[0077] S2: Preparation of Astragalus glycosides

[0078] S2.1: 10 parts by volume of 1 mmol / L kaempferol substrate, 100 mmol / L sucrose, 0.2 mmol / L UDP, 120 mmol / L Tris-HCl buffer (pH=7), 60 mmol / LMgCl, 5% DMSO, FCT and GmSUS enzyme solution were reacted at 35°C and 200 r / min for 1 h, 30 parts by volume of methanol were added to terminate the reaction, and the mixture was centrifuged at 12000 r / min for 10 min at 0°C to obtain the supernatant to obtain reaction solution I;

[0079] S2.2: The reaction solution I is centrifuged twice at 12000 r / min for 15 min, and the obtained supernatant is added to an LH-20 polysaccharide gel column, equilibrated and eluted with a 50% ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside.

[0080] S3: Preparation of Astragalus glycoside nanoalbumin

[0081] S3.1: Dissolve 1 part by volume of astragaloside in 10 parts by volume of deionized water to obtain an astragaloside aqueous solution, adjust the pH of the astragaloside aqueous solution to 5.5 with dilute hydrochloric acid, add EDC and NHS in a ratio of 1:2:1.2, and activate with stirring for 1 hour in a dark place;

[0082] S3.2: Mix albumin nanoparticles and astragalus glycoside in a mass ratio of 1:5, adjust the pH of the system to 7.5 with PBS solution, stir the reaction for 14 hours, wash 3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin.

[0083] S4: Preparation of Astragalus glycoside nanoalbumin eye drops

[0084] S4.1: 25 parts by mass of hydroxypropyl-β-cyclodextrin and 45 parts by volume of deionized water are placed in an environment of 60°C and magnetically stirred until dissolved, and then 0.2 parts by mass of mannitol are added to obtain a saturated aqueous solution;

[0085] S4.2: Weigh 1 part by mass of astragalus glycoside, slowly add it into a saturated aqueous solution at 60°C, sonicate for 65 minutes, and add water to 100 parts by volume to obtain an intermediate solution;

[0086] S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. The volume ratio of astragalus glycoside nano-albumin and the intermediate solution is 1:2. After filtering with a 0.22 μm filter membrane, astragalus glycoside nano-albumin eye drops are obtained.

[0087] Comparative Example 1:

[0088] Compared with Example 1, the difference of Comparative Example 1 is that, in step S3, albumin nanoparticles are not added, but hydroxypropyl-β-cyclodextrin is used, specifically: "S3.1: dissolving 1 part by volume of astragalus glycoside in 10 parts by volume of deionized water to obtain an astragalus glycoside aqueous solution, adjusting the pH of the astragalus glycoside aqueous solution to 5.5 with dilute hydrochloric acid, adding EDC and NHS in a ratio of 1:2:1.2 in sequence, and stirring and activating for 2 hours in a dark place;

[0089] S3.2: Mix hydroxypropyl-β-cyclodextrin and astragalus glycoside in a mass ratio of 1:5, adjust the pH of the system to 7.5 with PBS solution, stir the reaction for 12 hours, wash 3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside cyclodextrin inclusion complex. Replace the astragalus glycoside nanoalbumin in step S4 with the astragalus glycoside cyclodextrin inclusion complex, and the prepared eye drops are recorded as Comparative Example 1.

[0090] Comparative Example 2:

[0091] Compared with Example 1, the difference of Comparative Example 2 is that hydroxypropyl-β-cyclodextrin is not added in step S4, but all water is added, specifically: "S4.1: 45 parts by volume of deionized water are placed in an environment of 65°C and stirred with a constant temperature magnetic force until dissolved, and then 0.2 parts by mass of mannitol are added to obtain a saturated aqueous solution;

[0092] S4.2: Weigh 1 part by mass of astragalus glycoside, slowly add it into a saturated aqueous solution at 65°C, sonicate for 60 minutes, and add water to 100 parts by volume to obtain an intermediate solution;

[0093] S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. The volume ratio of astragalus glycoside nano-albumin to the intermediate solution is 1:2. After filtering with a 0.22 μm filter membrane, astragalus glycoside nano-albumin eye drops are obtained. The prepared eye drops are recorded as Comparative Example 2.

[0094] Eye irritation evaluation

[0095] The eye irritation of the comparative examples and the examples to rabbits was examined by the Draize eye irritation test, and the eye conditions of the rabbits after continuous administration were observed, as shown in Table 1.

[0096] Table 1: Evaluation of rabbit eye irritation

[0097]

[0098] It can be seen that Comparative Example 1 does not add albumin nanoparticles, which is irritating to the rabbit's eyes to a certain extent, causing the rabbit's eyes to become congested after the drops are dripped, while the embodiments do not have any congestion and no obvious secretions, proving that the rabbit's eyes adapt well to the embodiments and have low irritation. Comparative Example 2 does not add hydroxypropyl-β-cyclodextrin eye drops, so that astragalus glycosides exist freely in the solution and are easy to aggregate, which greatly increases the irritation to the rabbit's eyes. Therefore, obvious secretions and congestion appear in the rabbit's eyes, which is irritating to a certain extent.

[0099] In vitro release rate

[0100] 10 mL of the comparative example and the example were added to 100 mL of artificial tears respectively, and the mixture was shaken at a constant temperature of 37°C in a shaker in the dark. After sampling 3 mL every 1 h, an equal volume of isothermal blank artificial tears was added. The fluorescence intensity of the samples was measured by a fluorescence spectrophotometer (Ex = 466 nm, Em = 490 nm), and the cumulative release rate was calculated as shown in Table 2.

[0101] Table 2: Time and release rate chart

[0102]

[0103] It can be seen that the 5h release rate of the embodiment is 8.7-8.9%, and in the subsequent 10-60h, the release rates of the embodiment are 38%, 56%, 61% and 68%, which are all lower than the release rate of the comparative example, and at 40-60h, the release rate of the comparative example rises rapidly, from 63% to 78% and from 64% to 82%, respectively. This is because albumin nanoparticles are not used to couple astragalus glycoside in Comparative Example 1, which reduces the stability of astragalus glycoside, and cyclodextrin-encapsulated astragalus glycoside is more easily dissociated, thereby reducing the release rate. In Comparative Example 2, deionized water is used instead of cyclodextrin to carry astragalus glycoside, and the inclusion rate is further reduced. At 60h, astragalus glycoside is greatly released and agglomerated, so that the release rate of Comparative Example 2 is as high as 82%.

[0104] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A preparation process of a protein eye protection solution, characterized in that: The steps include: S1: Preparation of albumin nanoparticles Weigh bovine serum albumin, dissolve it in a PBS solution, add a composite emulsifier to obtain a solution to be emulsified, add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification, and then perform rotary evaporation, then dropwise add anhydrous ethanol under stirring until a colloidal solution is formed, add glutaraldehyde to the colloidal solution, continue stirring and solidifying, wash after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain albumin nanoparticles; S2: Preparation of Astragalus glycosides The enzyme solution of kaempferol substrate, sucrose, UDP, Tris-HCl buffer, MgCl, DMSO, FCT and GmSUS is reacted at 32-35°C and 200-250 r / min for 1-4 hours, methanol is added to terminate the reaction, and the supernatant is obtained by centrifugation to obtain reaction solution I, and the reaction solution I is centrifuged twice, and the obtained supernatant is added to an LH-20 dextran gel column, equilibrated and eluted with an ethanol solution, and the eluate is collected and freeze-dried to obtain astragaloside; The specific steps include: S2.1: 8-10 parts by volume of 1-2 mmol / L kaempferol substrate, 100-110 mmol / L sucrose, 0.2-0.3 mmol / L UDP, 100-120 mmol / L Tris-HCl buffer, 50-60 mmol / LMgCl, 5-6% DMSO, FCT and GmSUS enzyme solution were reacted at 32-35°C and 200-250 r / min for 1-4 h, 20-30 parts by volume of methanol were added to terminate the reaction, and the mixture was centrifuged at 12000-15000 r / min for 8-10 min at 0-4°C to obtain the supernatant to obtain reaction solution I; S2.2: centrifuge the reaction solution I twice, at 12000-15000 r / min for 10-15 min, add the obtained supernatant to an LH-20 polysaccharide gel column, balance and elute with 45-50% ethanol solution, collect the eluate, freeze-dry, and obtain astragaloside; S3: Preparation of Astragalus glycoside nanoalbumin Dissolve astragalus glycoside in deionized water to obtain an astragalus glycoside aqueous solution, adjust the pH of the astragalus glycoside aqueous solution to 4-5.5 with dilute hydrochloric acid, add EDC and NHS in sequence, stir and activate in the dark, mix the albumin nanoparticles with astragalus glycoside, wash after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin; S4: Preparation of Astragalus glycoside nanoalbumin eye drops Hydroxypropyl-β-cyclodextrin and deionized water were magnetically stirred at a constant temperature until dissolved, and mannitol was added to obtain a saturated aqueous solution. Astragalus glycoside was weighed and slowly added to the saturated aqueous solution. After ultrasound, water was added to 100 parts by volume to obtain an intermediate solution, and astragalus glycoside nanoalbumin was added and stirred at room temperature overnight. After filtering with a 0.22 μm filter membrane, astragalus glycoside nanoalbumin eye drops were obtained.

2. The preparation process of a protein eye protection solution according to claim 1, characterized in that: Step S1: preparing albumin nanoparticles, comprising the following steps: S1.1: Weigh bovine serum albumin and dissolve it in PBS solution. The solid-liquid ratio of bovine serum albumin to PBS solution is 1:(18-20). Adjust the pH of the system to 7.5-8, add a composite emulsifier with a concentration of 0.5-0.75%, and obtain an emulsified solution. S1.2: Add the solution to be emulsified into an ultrasonic machine for ultrasonic emulsification for 10-15 minutes, then transfer to a rotary evaporator for rotary evaporation for 10-15 minutes, and then add anhydrous ethanol dropwise under stirring until a colloidal solution is formed; S1.3: Add glutaraldehyde to the colloidal solution and continue stirring to solidify for 1-2 hours. After high-speed centrifugation, wash 1-3 times, collect the precipitate, and freeze-dry to obtain albumin nanoparticles.

3. The preparation process of a protein eye protection solution according to claim 2, characterized in that: Step S3 prepares astragalus glycoside nano-albumin, comprising the following steps: S3.1: Dissolve 1-5 parts by volume of astragaloside in 10-15 parts by volume of deionized water to obtain an astragaloside aqueous solution, adjust the pH of the astragaloside aqueous solution to 4-5.5 with dilute hydrochloric acid, add EDC and NHS in a ratio of 1: (1.5-2): (1.2-1.5) in sequence, and stir to activate for 1-2 hours in a dark place; S3.2: Mix albumin nanoparticles and astragalus glycoside in a mass ratio of 1: (4-5), adjust the pH of the system to 7-7.5 with PBS solution, stir the reaction for 12-14 hours, wash 1-3 times after high-speed centrifugation, collect the precipitate, and freeze-dry to obtain astragalus glycoside nanoalbumin.

4. The preparation process of a protein eye protection solution according to claim 3, characterized in that: Step S4 prepares astragalus glycoside nano-albumin eye drops, comprising the following steps: S4.1: Place 25-30 parts by mass of hydroxypropyl-β-cyclodextrin and 40-45 parts by volume of deionized water in an environment of 60-65°C with constant temperature magnetic stirring until dissolved, and then add 0.15-0.2 parts by mass of mannitol to obtain a saturated aqueous solution; S4.2: Weigh 0.2-1 parts by mass of astragalus glycoside, slowly add it to a saturated aqueous solution at 60-65°C, sonicate for 60-65 minutes, and add water to 100 parts by volume to obtain an intermediate solution; S4.3: Add astragalus glycoside nano-albumin and stir at room temperature overnight. Filter with 0.22 μm filter membrane to obtain astragalus glycoside nano-albumin eye drops.

5. The preparation process of a protein eye protection solution according to claim 4, characterized in that: In step S1, the composite emulsifier is prepared by mixing sodium caseinate and poloxamer in a volume ratio of 2:

1.

6. The preparation process of a protein eye protection solution according to claim 5, characterized in that: The pH of the Tris-HCl buffer in step S2 is 7.

7. The preparation process of a protein eye protection solution according to claim 6, characterized in that: The volume ratio of the colloidal solution to glutaraldehyde in step S1 is 1:(0.2-0.5).

8. The preparation process of a protein eye protection solution according to claim 7, characterized in that: In step S4, the volume ratio of astragalus glycoside nanoalbumin to the intermediate solution is 1:(2-3).

9. A protein eye protection liquid, which is prepared by the preparation process of the protein eye protection liquid according to any one of claims 1 to 8.

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