An eye care composition containing ergothioneine, its preparation method and applications

Abstract

The present invention provides an eye care composition containing ergothioneine, its preparation method and application, which relates to the technical field of medicine. The eye care composition containing ergothioneine provided by the present invention comprises the following components: ergothioneine, taurine, sodium hyaluronate, plant essential oil, terephthalic boric acid, an osmotic pressure regulator and water for injection. The preparation method is to mix sodium hyaluronate with water for injection to obtain solution A; mix solution A with taurine, ergothioneine and terephthalic boric acid, and adjust the pH to 5-7.5 to obtain solution B; mix solution B with plant essential oil, add an osmotic pressure regulator to adjust the osmotic pressure to 250-350 mOsm / (Kg·H2O), filter and sterilize, and subpackage to obtain the product. Through specific components and ratios, the present invention obtains an eye care composition containing ergothioneine with high stability, anti-inflammatory effect, ability to relieve eye fatigue, eye itching and dry eyes, and good moisturizing effect.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to an eye care composition containing ergothioneine, a preparation method thereof, and an application thereof. Background Art

[0002] The frequent use of electronic products can easily cause visual fatigue, dryness, soreness and swelling in the eyes, resulting in the weakening of the eye's antioxidant defense system, leading to decreased vision, blurred vision, and further causing other eye diseases. Eye wash is added with natural active substances, and is formulated into a solution adapted to the human body with purified water or distilled water and inorganic salt substances, which can clean and maintain the eyes, play a good role in relieving eye fatigue, maintaining eye cleanliness, slowing down some eye diseases, repairing some minor physical damages in the eyes, etc. At the same time, it can stimulate the microvascular circulation of the eyes, meet the nutrient needs of the eye skin, and improve the dark circle problems caused by fatigue, staying up late, and poor sleep, and is deeply liked by consumers.

[0003] Chinese Patent CN 116889599 A discloses a blueberry eye wash and a preparation method thereof in the technical field of eye wash. Every 1000 mL of the eye wash is prepared from the following raw materials: 60 - 80 g of blueberries, 80 - 100 g of folium isatidis, 20 - 40 g of mulberry leaves, 2 - 3 g of compound vitamins, and 0.2 - 0.25 g of sodium hyaluronate. The compound vitamins are any two of vitamin B6, vitamin B12, vitamin A, and vitamin E. The eye wash is prepared by mixing blueberries, folium isatidis, mulberry leaves, compound vitamins, and sodium hyaluronate. The raw material cost is low. By adopting the local administration method, it can directly reach the action site, with a fast absorption speed and obvious use effect. However, its effect only tends to soothe dryness and does not have the effects of anti-fatigue, anti-inflammatory, etc.

[0004] Chinese Patent CN 114099551 B discloses a vision fatigue-relieving eye lubricating composition, which contains main components such as glucosylglycerol, zinc sulfate, tea polyphenols, taurine, dipotassium glycyrrhizinate, menthol, borneol, etc. It can be used in eye spray and eye wash aqueous products, which not only has the effects of relieving visual fatigue, eye itching, eye perimeter dryness, eye dryness, and improving eyesight, but also has good moisturizing effect, is natural, safe and non-irritating, and has good application prospects. However, its components are relatively complex and no stability test has been carried out.

[0005] Chinese Patent CN 114949035 A discloses an eye wash for preventing dry eye and its preparation method, which is prepared from the following raw materials: 4 - 7 mg of taurine, 3 - 7 mg of borneol, 60 - 80 g of cornflower, 80 - 100 g of folium isatidis, 20 - 40 g of mint, 20 - 30 g of vitamin A, 6 - 8 mL of honey, and 6 - 8 mL of pearl liquid. It provides a safe, effective and convenient ophthalmic pharmaceutical preparation for the clinical treatment of dry eye. The utility of this product is only for dry eye, which is relatively single and cannot meet the needs of consumers with various eye symptoms.

[0006] On the one hand, most common eye wash and care products on the market can only play a role in cleaning the eyes, and their ingredients are mostly chemical drugs, which have great irritation to the eyes and will produce side effects after long-term use. On the other hand, eye washes or eye drops with functions such as anti-inflammatory, anti-allergic or vasoconstriction usually use drugs such as antibiotics, which have strong side effects and will cause discomfort to normal people and have strong irritation to the eyeballs. Moreover, the efficacy of existing eye washes is usually relatively single, only effective for a certain eye symptom and cannot meet the needs of consumers. Therefore, it is urgent to develop an eye wash with natural and mild ingredients, multiple functions and good stability.

[0007] Based on this, the researchers of the present invention have developed an eye care composition containing ergothioneine and its preparation method. By optimizing the formula and ratio, an eye wash with high stability, anti-inflammatory effect, relieving visual fatigue, eye itching and eye dryness, and good moisturizing effect is obtained. Summary of the Invention

[0008] The present invention aims at the above problems and provides an eye care composition containing ergothioneine and its preparation method, which includes ergothioneine, taurine, sodium hyaluronate, plant essential oil, bacteriostatic agent and osmotic pressure regulator. Through the synergistic effect of each component and a specific ratio, an eye care composition containing ergothioneine with high stability, anti-inflammatory effect, relieving visual fatigue, eye itching and eye dryness, and good moisturizing effect is obtained.

[0009] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0010] On the one hand, the present invention provides an eye care composition containing ergothioneine, which includes the following components: ergothioneine, taurine, sodium hyaluronate, plant essential oil, terephthalic acid boric acid, osmotic pressure regulator and water for injection.

[0011] Preferably, the mass ratio of the ergothioneine, taurine and terephthalic acid boric acid is 70 - 120:190 - 210:1; further preferably, the mass ratio of the ergothioneine, taurine and terephthalic acid boric acid is 90 - 100:190 - 250:1.

[0012] Preferably, the mass ratio of taurine, sodium hyaluronate and plant essential oil is 10-25:1-10:1; more preferably, the mass ratio of taurine, sodium hyaluronate and plant essential oil is 16-20:4-7:1.

[0013] Preferably, the ergot thioneine-containing eye care composition comprises the following components in parts by weight: 0.01-1 part of ergot thioneine, 0.1-5 parts of taurine, 0.05-0.4 part of sodium hyaluronate, 0.03-0.06 part of plant essential oil, 0.001-0.004 part of terephthalic boric acid, and 0.1-0.5 part of osmotic pressure regulator, and the balance is made up to 100 parts by weight with water for injection.

[0014] Preferably, the ergot thioneine-containing eye care composition comprises the following components in parts by weight: 0.25-0.42 part of ergot thioneine, 0.6-1.2 parts of taurine, 0.2-0.4 part of sodium hyaluronate, 0.03-0.06 part of plant essential oil, 0.001-0.004 part of terephthalic boric acid, and 0.1-0.5 part of osmotic pressure regulator, and the balance is made up to 100 parts by weight with water for injection.

[0015] More preferably, the ergot thioneine-containing eye care composition comprises the following components in parts by weight: 0.3-0.4 part of ergot thioneine, 0.65-1.0 part of taurine, 0.22-0.38 part of sodium hyaluronate, 0.035-0.055 part of plant essential oil, 0.002-0.004 part of terephthalic boric acid, and 0.2-0.5 part of osmotic pressure regulator, and the balance is made up to 100 parts by weight with water for injection.

[0016] Even more preferably, the ergot thioneine-containing eye care composition comprises the following components in parts by weight: 0.3-0.35 part of ergot thioneine, 0.7-0.8 part of taurine, 0.22-0.3 part of sodium hyaluronate, 0.04-0.05 part of plant essential oil, 0.003-0.004 part of terephthalic boric acid, and 0.3-0.5 part of osmotic pressure regulator, and the balance is made up to 100 parts by weight with water for injection.

[0017] Preferably, the ergot thioneine-containing eye care composition further comprises a pH regulator.

[0018] Preferably, the pH of the ergot thioneine-containing eye care composition is 5-7.5.

[0019] Preferably, the pH regulator is selected from at least one of citrate buffer solution and borate buffer solution; more preferably, the pH regulator is citrate buffer solution.

[0020] Preferably, the plant essential oil is selected from at least one of rose essential oil, chamomile essential oil, and peppermint essential oil; further preferably, the plant essential oil is selected from at least one of chamomile essential oil and peppermint essential oil; more preferably, the plant essential oil is chamomile essential oil and peppermint essential oil.

[0021] Preferably, the mass ratio of the chamomile essential oil to the peppermint essential oil is 1-6:2-4; further preferably, the mass ratio of the chamomile essential oil to the peppermint essential oil is 4-6:2-3; more preferably, the mass ratio of the chamomile essential oil to the peppermint essential oil is 5:2.

[0022] Preferably, the osmotic pressure regulator is selected from at least one of sodium chloride, potassium chloride, sorbitol, borax, and boric acid; further preferably, the osmotic pressure regulator is selected from at least one of sodium chloride and potassium chloride; more preferably, the osmotic pressure regulator is sodium chloride.

[0023] On the other hand, the present invention also provides a preparation method of the above-mentioned eye care composition containing ergothioneine, which comprises the following steps:

[0024] S1: Mix sodium hyaluronate with water for injection to obtain solution A;

[0025] S2: Mix solution A with taurine, ergothioneine, and terephthalic acid, and adjust the pH to 5-7.5 to obtain solution B;

[0026] S3: Mix solution B with the plant essential oil, add an osmotic pressure regulator to adjust the osmotic pressure to 250-350 mOsm / (Kg·H2O), filter and sterilize, and subpackage to obtain the product.

[0027] Preferably, the temperature of the mixing in S1 is 75-85°C; further preferably, the temperature of the mixing is 80°C.

[0028] Preferably, the mixing in S1 is stirring.

[0029] Preferably, the temperature of the mixing in S2 is 35-45°C; further preferably, the temperature of the mixing in S2 is 35°C.

[0030] Preferably, the mixing in S2 can be selected from at least one of stirring and ultrasonic treatment; further preferably, the mixing in S2 is ultrasonic treatment.

[0031] Preferably, the temperature of the mixing in S3 is 30-35°C; further preferably, the temperature of the mixing in S3 is 30°C.

[0032] Preferably, the mixing in S3 can be selected from at least one of stirring and ultrasonic treatment; further preferably, the mixing in S3 is stirring.

[0033] On the other hand, the present invention also provides the application of the above-mentioned eye care composition containing ergothioneine in eye care.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] 1. In the research of the present invention, it is found that ergothioneine, taurine and terephthalic boric acid have a synergistic effect in enhancing the antioxidant effect. Especially under the specific ratio of the technical solution of the present invention, the antioxidant property of the eye care composition containing ergothioneine is significantly improved;

[0036] 2. In the research of the present invention, it is found that taurine, sodium hyaluronate and plant essential oils have a synergistic effect in soothing the eyes and relieving fatigue. Especially under the specific ratio of the technical solution of the present invention, the anti-fatigue effect of the eye care composition containing ergothioneine is significantly improved;

[0037] 3. In the research of the present invention, it is found that taurine and terephthalic boric acid have a synergistic effect in enhancing the antibacterial effect. Especially under the specific ratio of the technical solution of the present invention, the antibacterial effect of the eye care composition containing ergothioneine is significantly improved;

[0038] 4. Through the specific components and ratios of the technical solution of the present invention, an eye care composition containing ergothioneine with high stability, anti-inflammatory, relieving visual fatigue, eye itching, eye dryness, and good moisturizing effect is obtained. Detailed Embodiments

[0039] In order to make the technical means, creative features, achieved purposes and functions of the present invention easy to understand, the present invention will be further clarified below with reference to specific embodiments. However, the following embodiments are only the preferred embodiments of the present invention, not all of them. Based on the embodiments in the embodiments, other embodiments obtained by those skilled in the art without creative efforts all belong to the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all ordinary commercially available products, and no specific limitation is made on their sources. The technical and scientific terms used in the embodiments have the same meanings as those commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0040] Example 1

[0041] 1) Components

[0042] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of terephthalic boric acid, and 0.44 parts of sodium chloride, and the injection water is made up to 100 parts by weight.

[0043] 2) Preparation

[0044] S1: Dissolve sodium hyaluronate and water for injection by stirring at 80 °C to obtain Solution A;

[0045] S2: Ultrasonicate Solution A with taurine, ergothioneine and terephthalic boric acid at 35 °C for 20 min, add an appropriate amount of citrate buffer solution, and adjust the pH to 5 - 7.5 to obtain Solution B;

[0046] S3: Stir Solution B and plant essential oil at 30 °C, mix evenly, add sodium chloride to adjust the osmotic pressure to 310 mOsm / (Kg·H2O), perform filtration sterilization, and subpackage to obtain the product.

[0047] Example 2

[0048] Compared with Example 1, only the components are changed, specifically:

[0049] 1) Components

[0050] By weight, it contains the following components: 0.12 parts of ergothioneine, 0.19 parts of taurine, 0.05 parts of sodium hyaluronate, 0.021 parts of chamomile essential oil, 0.009 parts of peppermint essential oil, 0.001 parts of terephthalic boric acid and 0.5 parts of sodium chloride, and water for injection makes up 100 parts by weight.

[0051] The remaining operations are the same as those in Example 1.

[0052] Example 3

[0053] Compared with Example 1, only the components are changed, specifically:

[0054] 1) Components

[0055] By weight, it contains the following components: 0.4 parts of ergothioneine, 1.0 parts of taurine, 0.4 parts of sodium hyaluronate, 0.043 parts of chamomile essential oil, 0.017 parts of peppermint essential oil, 0.004 parts of terephthalic boric acid and 0.1 parts of sodium chloride, and water for injection makes up 100 parts by weight.

[0056] The remaining operations are the same as those in Example 1.

[0057] Comparative Example 1

[0058] Compared with Example 1, only the components are changed, using taurine instead of ergothioneine, specifically:

[0059] By weight, it contains the following components: 1.1 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of terephthalic boric acid and 0.44 parts of sodium chloride, and water for injection makes up 100 parts by weight.

[0060] The remaining operations are the same as those in Example 1.

[0061] Comparative Example 2

[0062] Compared with Example 1, only the components are changed, and ergothioneine is replaced with terephthalic acid diboronic acid, specifically:

[0063] By weight, it contains the following components: 0.3536 parts of terephthalic acid diboronic acid, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, and 0.44 parts of sodium chloride, and the injection water is added to make up 100 parts by weight.

[0064] The remaining operations are the same as those in Example 1.

[0065] Comparative Example 3

[0066] Compared with Example 1, only the components are changed, and terephthalic acid diboronic acid is replaced with taurine, specifically:

[0067] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.7536 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, and 0.44 parts of sodium chloride, and the injection water is added to make up 100 parts by weight.

[0068] The remaining operations are the same as those in Example 1.

[0069] Comparative Example 4

[0070] Compared with Example 1, only the components are changed, and terephthalic acid diboronic acid is replaced with dipotassium glycyrrhizinate, specifically:

[0071] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of dipotassium glycyrrhizinate, and 0.44 parts of sodium chloride, and the injection water is added to make up 100 parts by weight.

[0072] The remaining operations are the same as those in Example 1.

[0073] Comparative Example 5

[0074] Compared with Example 1, only the components are changed, and terephthalic acid diboronic acid is replaced with p-hydroxybenzoate, specifically:

[0075] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of p-hydroxybenzoate, and 0.44 parts of sodium chloride, and the injection water is added to make up 100 parts by weight.

[0076] The remaining operations are the same as those in Example 1.

[0077] Comparative Example 6

[0078] Compared with Example 1, only the components are changed, and benzalkonium chloride is used instead of p-phenylenediboronic acid. Specifically:

[0079] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of benzalkonium chloride, and 0.44 parts of sodium chloride, and the balance is made up to 100 parts by weight with water for injection.

[0080] The remaining operations are the same as those in Example 1.

[0081] Comparative Example 7

[0082] Compared with Example 1, only the components are changed, and thimerosal is used instead of p-phenylenediboronic acid. Specifically:

[0083] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of thimerosal, and 0.44 parts of sodium chloride, and the balance is made up to 100 parts by weight with water for injection.

[0084] The remaining operations are the same as those in Example 1.

[0085] Comparative Example 8

[0086] Compared with Example 1, only the components are changed, and sodium hyaluronate is used instead of taurine. Specifically:

[0087] By weight, it contains the following components: 0.35 parts of ergothioneine, 1 part of sodium hyaluronate, 0.03 parts of chamomile essential oil, 0.012 parts of peppermint essential oil, 0.0036 parts of p-phenylenediboronic acid, and 0.44 parts of sodium chloride, and the balance is made up to 100 parts by weight with water for injection.

[0088] The remaining operations are the same as those in Example 1.

[0089] Comparative Example 9

[0090] Compared with Example 1, only the components are changed, and sodium hyaluronate is used instead of plant essential oil. Specifically:

[0091] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.292 parts of sodium hyaluronate, 0.0036 parts of p-phenylenediboronic acid, and 0.44 parts of sodium chloride, and the balance is made up to 100 parts by weight with water for injection.

[0092] The remaining operations are the same as those in Example 1.

[0093] Comparative Example 10

[0094] Compared with Example 1, only the components were changed, and chamomile essential oil was used instead of peppermint essential oil. Specifically:

[0095] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.042 parts of chamomile essential oil, 0.0036 parts of terephthalic boric acid, and 0.44 parts of sodium chloride, and the injection water is made up to 100 parts by weight.

[0096] The remaining operations are the same as those in Example 1.

[0097] Comparative Example 11

[0098] Compared with Example 1, only the components were changed, and the plant essential oil was removed. Specifically:

[0099] By weight, it contains the following components: 0.35 parts of ergothioneine, 0.75 parts of taurine, 0.25 parts of sodium hyaluronate, 0.0036 parts of terephthalic boric acid, and 0.44 parts of sodium chloride, and the injection water is made up to 100 parts by weight.

[0100] The remaining operations are the same as those in Example 1.

[0101] Comparative Example 12

[0102] Compared with Example 1, only the component ratio was changed. Specifically:

[0103] By weight, it contains the following components: 0.1 part of ergothioneine, 1.5 parts of taurine, 0.05 part of sodium hyaluronate, 0.11 part of chamomile essential oil, 0.01 part of peppermint essential oil, 0.0056 parts of terephthalic boric acid, and 0.060 parts of sodium chloride, and the injection water is made up to 100 parts by weight.

[0104] The remaining operations are the same as those in Example 1.

[0105] Comparative Example 13

[0106] Compared with Example 1, only the preparation method was changed. Specifically:

[0107] The components are the same as those in Example 1;

[0108] Preparation:

[0109] Stir and mix ergothioneine, taurine, sodium hyaluronate, terephthalic boric acid, chamomile essential oil, peppermint essential oil and injection water, add an appropriate amount of citrate buffer solution, adjust the pH to 5 - 7.5, add sodium chloride to adjust the osmotic pressure to 310 mOsm / (Kg·H2O), filter and sterilize, and subpackage to obtain.

[0110] Test Example 1

[0111] In vitro antioxidant experiment

[0112] Excessive production of ROS free radicals caused by hypoxia during daily wearing of contact lenses, especially the accumulation of superoxide anions (O 2 ). Now, antioxidant tests are carried out on the ergothioneine-containing eye care compositions prepared in Examples 1-3 and Comparative Examples 1-13. Superoxide anions are determined by the pyrogallol autoxidation method, the hydroxyl radical scavenging experiment adopts the Fenton colorimetric method, and the DPPH radical scavenging method is the most widely used method for indirectly detecting the antioxidant capacity of the tested substances. The antioxidant performance of the tested substances is judged by detecting the change in absorbance. The results are shown in Tables 1-3.

[0113] Table 1. O 2 Inhibition rate results

[0114] Example <![CDATA[O 2 Inhibition rate / %]]> Hydroxyl radical scavenging rate / % DPPH inhibition rate / % Example 1 99.5 65.4 99.4 Example 2 99.2 63.7 99.0 Example 3 99.3 62.5 99.1 Comparative Example 1 34.5 24.3 43.6 Comparative Example 2 33.6 20.9 40.9 Comparative Example 3 52.1 30.2 64.3 Comparative Example 4 38.2 22.1 42.2 Comparative Example 5 37.4 20.7 44.7 Comparative Example 6 31.9 25.7 41.6 Comparative Example 7 35.2 21.4 40.2 Comparative Example 8 47.1 29.8 63.7 Comparative Example 9 54.8 32.5 63.1 Comparative Example 10 61.5 34.7 67.2 Comparative Example 11 52.4 33.6 61.8 Comparative Example 12 31.6 20.3 43.4 Comparative Example 13 82.9 58.5 79.5

[0115] As shown in Table 1, the ergothioneine-containing eye care composition prepared by the technical solution of the present invention has good antioxidant effects, significantly superior to the comparative examples, and ergothioneine, taurine and terephthalic boric acid have a synergistic effect in enhancing the antioxidant effect. Especially at the specific ratio of the technical solution of the present invention, the antioxidant property of the ergothioneine-containing eye care composition is significantly improved.

[0116] Test Example 2

[0117] Cytotoxicity test

[0118] The corneal stromal cell suspension with a concentration of 1×10 5 cells / mL was added to a 96-well plate at 100 μL / well, and cultured in an incubator containing 5% CO 2 , 37 °C for 24 hours, and then the culture medium was aspirated. Then, the ergothioneine-containing eye care compositions prepared in Examples 1-3 and Comparative Examples 1-13 were added to the 96-well plate at a ratio of 1:10 with the complete culture medium, and the cells were cultured for another 24 h, and then cultured in the incubator for another 24 hours. Positive (10% DMSO), negative (complete cell culture medium) control groups and the ergothioneine-containing eye care compositions prepared in Examples 1-3 and Comparative Examples 1-13 were all set with 6 replicates. Add 20 μL / well of MTT and incubate for another 3 h. Then add 100 μL of DMSO, shake for 10 min, and measure the absorbance at a wavelength of 570 nm on an enzyme-linked immunosorbent assay (ELISA) reader. The cell viability was calculated as follows:

[0119] Cell viability = (OD value of the test substance well - OD value of the blank control well) / OD value of the negative control well.

[0120] The results are shown in Table 2.

[0121] Table 2. Results of cytotoxicity test

[0122]

[0123] From the data in Table 2, it can be seen that the survival rate of corneal stromal cells of all products at 24 h is above 99%, and the survival rate of corneal stromal cells at 72 h is above 98%, indicating that the eye care compositions containing ergothioneine prepared by the formulations of the examples and the comparative examples are safe and non-toxic to corneal stromal cells.

[0124] Test Example 3

[0125] Antibacterial effect

[0126] Using Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans as test bacteria, the antibacterial effects of the eye care compositions containing ergothioneine prepared in Examples 1-3 and Comparative Examples 1-13 against the 4 strains were tested.

[0127] Prepare the bacterial suspension for the experiment. Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and Candida albicans are cultured in agar medium. Add an appropriate amount of 0.9% sterile sodium chloride solution to elute the culture on the agar surface, and transfer the bacterial suspension to a sterile test tube. Dilute it with 0.9% sterile sodium chloride solution to make a bacterial suspension with about 108 cfu of bacteria per 1 mL. Take 0.5 mL of the test bacterial suspension in a test tube, then add 0.5 mL of organic interfering substance (3% bovine serum albumin solution), mix well, place it in a water bath at 20°C ± 1°C for 5 min, and then use a sterile pipette to respectively pipette 4.0 mL of the eye care compositions containing ergothioneine prepared in Examples 1-3 and Comparative Examples 1-13 into it, mix quickly and start timing immediately.

[0128] After the test bacteria interact with the samples of each example and comparative example for each predetermined time, respectively pipette 0.5 mL of the test bacteria and sample mixture into 4.5 mL of sterilized neutralizer, and mix well. After the test bacteria and sample mixture in each tube act with the neutralizer for 10 min, respectively pipette 1.0 mL of the sample solution, and determine the number of viable bacteria according to the viable bacteria culture counting method. It is sufficient to inoculate 2 petri dishes with each tube of sample solution. If there are more colonies growing on the petri dish, serial 10-fold dilutions can be carried out and then viable bacteria culture counting can be performed. At the same time, use the dilution solution instead of the sample for parallel tests as a control.

[0129] All test samples are cultured in an incubator at 37°C. Observe the final results after culturing the bacterial propagules for 48 h; observe the final results after culturing the bacterial spores for 72 h. The test is repeated 3 times, calculate the number of viable bacteria in each group, and then calculate the antibacterial rate according to the following formula:

[0130] Bacteriostatic rate = (number of viable bacteria in the control group - number of viable bacteria in the test group) / number of viable bacteria in the control group × 100%. The results are shown in Table 3 below.

[0131] Table 3. Bacteriostatic effect

[0132]

[0133]

[0134] As shown in Table 3, the eye care composition containing ergothioneine prepared by the technical solution of the present invention has a good bacteriostatic effect, which is significantly better than that of the comparative example. Moreover, taurine and terephthalic boric acid have a synergistic effect in enhancing the antioxidant effect. Especially under the specific ratio of the technical solution of the present invention, the bacteriostatic effect of the eye care composition containing ergothioneine is significantly improved.

[0135] Test Example 4

[0136] In vivo experiment:

[0137] 1. Retention time experiment

[0138] Add 2% sodium fluorescein to the eye care compositions containing ergothioneine prepared in Examples 1 - 3 and Comparative Examples 1 - 13, and mix well. Using New Zealand white rabbits as experimental subjects, randomly divide them into 16 groups, with 6 rabbits in each group. There are no significant differences in the age and weight of the rabbits in each group. Pull open one eyelid of the rabbit's eye, and drip 30 μL of the above different eye care compositions containing ergothioneine into the rabbit's eye respectively, and manually close the rabbit's eye for 10 s. Then, irradiate the eye with an ultraviolet lamp every 2 min, observe the intensity of the continuous fluorescent layer on the corneal surface, and record the time when the continuous fluorescent layer disappears as the eye retention time. Measure 3 times continuously, and take the average value as the eye retention time. The results are shown in Table 4.

[0139] Table 4. Eye retention time

[0140]

[0141] From the data in Table 4, it can be seen that compared with the eye care composition containing ergothioneine prepared in the comparative example, the retention time of the eye care composition containing ergothioneine prepared by the technical solution of the present invention is significantly increased, indicating that the specific components, ratio and preparation method of the technical solution of the present invention improve the bioavailability of the eye care composition containing ergothioneine.

[0142] 2. Animal experiment for dry eye

[0143] 1. Experimental materials

[0144] 1.1 Animals:

[0145] 95 SPF-grade male BALB / c mice, 6 - 8 weeks old, with body weight of (21 ± 3) g. After the animals were transferred to the animal house, they were adaptively fed for 1 week with free access to food and water. The temperature was (24 ± 2) °C and the humidity was 47%.

[0146] 1.2 Method for establishing an animal model of dry eye

[0147] Five mice were taken as the blank group and were injected with normal saline daily. The remaining 90 mice were modeled by instilling 0.2% benzalkonium chloride solution into the eyes, 5 μL / eye each time, once a day, to establish a benzalkonium chloride-induced dry eye mouse model. After 6 weeks of instillation, the tear secretion volume and tear breakup time (BUT) were detected to judge the success of the model establishment in the mice, and the unqualified mice were excluded.

[0148] Modeling results of 90 mice: 85 mice were successfully modeled and 5 mice failed to be modeled. The 85 successfully modeled mice were randomly divided into groups of 5 each, with a total of 17 groups (16 experimental groups, including Examples 1 - 3 and Comparative Examples 1 - 13), and one of them was the model group.

[0149] 1.3 Administration method:

[0150] In the blank group, 0.2 mL of sterilized normal saline was instilled into the left and right eyes of each mouse twice a day.

[0151] In experimental groups 1 - 16 (corresponding to Examples 1 - 3 and Comparative Examples 1 - 13 respectively), 0.2 mL of the eye care composition containing ergothioneine prepared in Examples 1 - 3 and Comparative Examples 1 - 13 was instilled into the left and right eyes of each mouse twice a day.

[0152] In the model group, 0.2 mL of sterilized normal saline was instilled into the left and right eyes of each mouse twice a day.

[0153] For 28 consecutive days, the observation indicators were recorded.

[0154] 1.4 Observation indicators:

[0155] 1) Tear secretion volume

[0156] After modeling, the tear secretion volume of the mice was detected using a tear secretion test strip (Schirmer I test, SIT) to judge whether the model was established. After drug intervention, the tear secretion volume of the mice was detected again using a tear secretion test strip to judge the effect of the intervention drug on the tear secretion of the mice.

[0157] 2) Measurement of tear breakup time

[0158] After modeling, the BUT of the mice was observed by fluorescein staining to judge whether the model was established. After drug intervention, the BUT was detected again to judge the effect of the intervention drug on the tear secretion of the mice.

[0159] 3) Detection of the expression levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor α (TNF-α) in the corneal tissues of mice in each group by immunofluorescence method: After 4 weeks of drug administration, the mice were overdosed with anesthesia and sacrificed. After enucleating the eyeballs, they were quickly fixed in 4% paraformaldehyde for later immunofluorescence detection. The fixed eyeballs were taken out, dehydrated by an automatic paraffin dehydrator, then embedded, sectioned, dewaxed, and baked. The antigens were repaired by heat-induced epitope retrieval method. After blocking with 5% BSA and washing with PBS, the primary antibody and the secondary antibody labeled with Alexa Fluor 488 were added dropwise. Finally, it was observed under a light microscope, and the average optical density values in 4 random fields of view were calculated for statistics.

[0160] 1.5 Statistical analysis:

[0161] Analysis and processing were performed using SPSS 16.0 statistical software. The data were expressed as (x±s), and one-way analysis of variance was used for comparison between groups. The results are shown in Table 5.

[0162] Table 5. Results of animal experiments on dry eye

[0163]

[0164] Note: Test Examples 1-3 correspond to Examples 1-3, and Test Examples 4-16 correspond to Comparative Examples 4-16;

[0165] Compared with the model group, **P<0.01; *P<0.05.

[0166] From the data in Table 5, it can be seen that compared with the eye care composition containing ergothioneine prepared in the comparative example, the anti-inflammatory and moisturizing properties of the eye care composition containing ergothioneine prepared by the technical solution of the present invention are significantly improved, indicating that the specific components, ratios, and preparation methods of the technical solution of the present invention improve the anti-inflammatory performance and soothing performance of the eye care composition containing ergothioneine, etc.

[0167] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than a limitation on the protection scope of the present invention. Any simple modification or equivalent replacement of the technical solution of the present invention by those of ordinary skill in the art does not depart from the essence and scope of the technical solution of the present invention.

Claims

1. An eye care composition containing ergothioneine, characterized in that, By weight parts, it comprises the following components: ergothioneine 0.01 - 1 part, taurine 0.1 - 5 parts, sodium hyaluronate 0.05 - 0.4 parts, plant essential oil 0.03 - 0.06 parts, terephthalic diboric acid 0.001 - 0.004 parts and an osmotic pressure regulator 0.1 - 0.5 parts, and the balance is made up to 100 weight parts with injection water; the mass ratio of the ergothioneine, taurine and terephthalic diboric acid is 70 - 120:190 - 250:

1.

2. The ergothioneine-containing eye care composition according to claim 1, characterized in that, The mass ratio of the taurine, sodium hyaluronate and plant essential oil is 10 - 25:1 - 10:

1.

3. The ergothioneine-containing eye care composition according to claim 1, wherein By weight parts, it comprises the following components: ergothioneine 0.25 - 0.42 parts, taurine 0.6 - 1.2 parts, sodium hyaluronate 0.2 - 0.4 parts, plant essential oil 0.03 - 0.06 parts, terephthalic diboric acid 0.001 - 0.004 parts and an osmotic pressure regulator 0.1 - 0.5 parts, and the balance is made up to 100 weight parts with injection water.

4. The ergothioneine-containing eye care composition according to claim 3, wherein By weight parts, it comprises the following components: ergothioneine 0.3 - 0.35 parts, taurine 0.7 - 0.8 parts, sodium hyaluronate 0.22 - 0.3 parts, plant essential oil 0.04 - 0.05 parts, terephthalic diboric acid 0.003 - 0.004 parts and an osmotic pressure regulator 0.3 - 0.5 parts, and the balance is made up to 100 weight parts with injection water.

5. The eye care composition containing ergothioneine according to claim 1, characterized in that, The described eye care composition containing ergothioneine further comprises a pH regulator.

6. The ergothioneine-containing eye care composition according to claim 5, wherein The pH of the described eye care composition containing ergothioneine is 5 - 7.

5.

7. The ergothioneine-containing eye care composition according to claim 5, characterized in that, The described pH regulator is selected from at least one of citrate buffer solution and borate buffer solution.

8. The eye care composition containing ergothioneine according to claim 1, characterized in that, The described plant essential oil is selected from at least one of rose essential oil, chamomile essential oil and peppermint essential oil.

9. The ergothioneine-containing eye care composition according to claim 8, characterized in that, The described plant essential oil is selected from at least one of chamomile essential oil and peppermint essential oil.

10. The ergothioneine-containing eye care composition according to claim 9, characterized in that, The described plant essential oil is chamomile essential oil and peppermint essential oil.

11. The ergothioneine-containing eye care composition according to claim 10, characterized in that, The mass ratio of the described chamomile essential oil and peppermint essential oil is 1 - 6:2 - 4.

12. The ergothioneine-containing eye care composition according to claim 1, wherein The described osmotic pressure regulator is selected from at least one of sodium chloride, potassium chloride, sorbitol, borax and boric acid.

13. A method for preparing the eye care composition containing ergothioneine according to any one of claims 1-12, characterized in that, It comprises the following steps: S1: Mix sodium hyaluronate with injection water to obtain solution A; S2: Mix solution A with taurine, ergothioneine and terephthalic diboric acid, and adjust the pH to 5 - 7.5 to obtain solution B; S3: Mix solution B with plant essential oil, add an osmotic pressure regulator to adjust the osmotic pressure to 250 - 350 mOsm / (Kg·H2O), carry out filtration and sterilization, and subpackage to obtain the product.

14. According to the preparation method described in claim 13, wherein the temperature of the mixing in S1 is 75 - 85 °C; the temperature of the mixing in S2 is 35 - 45 °C; the temperature of the mixing in S3 is 30 - 35 °C.

15. Use of the eye care composition containing ergothioneine according to any one of claims 1 - 12 in the preparation of eye care products.

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