A multi-effect composition containing ergothioneine, its preparation method and application in eye products

Abstract

The application belongs to the technical field of medicine, and particularly relates to a multiple-effect composition containing ergothioneine, a preparation method thereof and application of the multiple-effect composition in eye products. The multiple-effect composition contains ergothioneine, tromethamine and sodium hyaluronate in a mass ratio of 0.5-10:1-5:0.5-3. The application provides a multiple-effect composition containing ergothioneine. The tromethamine in the composition can protect the stability of ergothioneine under light and high-temperature conditions; and ergothioneine can protect the activity of macromolecular sodium hyaluronate. The synergistic compatibility of ergothioneine, sodium hyaluronate and tromethamine can significantly improve the moisturizing effect and eye-protecting effect of the composition. The multiple-effect composition containing ergothioneine provided by the application has the effects of preventing cataract and long-acting eye moistening, and has good stability.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a multifunctional composition containing ergothioneine, its preparation method, and its application in eye products. Background Technology

[0002] Mobile phone and computer screens emit blue light. Prolonged direct viewing of electronic screens leads to the accumulation of blue light energy, which can penetrate the cornea and lens to reach the retina. The main harms to the eyes include eye strain, dry eye syndrome, and pathological damage (including myopia, macular degeneration, and cataracts). Most existing eye drops contain moisturizing, lubricating, and anti-inflammatory ingredients, which can alleviate eye strain and discomfort to some extent. Their active ingredients are mostly phosphate esters, cellulose derivatives, and antibacterial and preservative quaternary ammonium salts. However, long-term use can lead to corneal lesions such as corneal calcification in some patients, premature onset of cataracts and presbyopia, and worsening of myopia. (The last sentence is a repetition of the previous one and can be omitted.)

[0003] For example, Chinese invention patent publication number CN114748611A discloses a composition with antioxidant, moisturizing and lubricating effects and its application. The composition includes, by mass percentage: 0.01%-1% collagen, 0.1%-5% ectoine, 0.005%-0.1% ergothioneine, 0.01%-1% moisturizer, 0.01%-10% buffer, 0.01%-2% osmotic pressure regulator, 0.001%-5% surfactant and 0.05%-5% chelating agent. In addition to being used as a lubricant when wearing contact lenses, this composition can also be used as an eye drop to relieve dryness and discomfort in the eyes. However, the composition is complex and contains many chemical components, which is not conducive to long-term use in the eyes, and the long-term moisturizing effect is not good.

[0004] Another Chinese invention patent, CN119587463A, discloses an eye drop containing L-ergothioneine and its preparation method. The eye drop comprises 0.0015-15 parts by weight of L-ergothioneine, 0.001-0.01 parts by weight of antibacterial agent, 0.05-15 parts by weight of buffer, and physiological saline to make up to 100 parts by weight. The pH of the eye drop is 5.0-7.0. The eye drop containing L-ergothioneine prepared by this invention can prevent and reduce lens protein turbidity. However, the effective ingredient of the eye drop is singular and cannot meet the needs of various eye symptoms.

[0005] Another Chinese invention patent publication, CN115068595A, discloses an ophthalmic anti-radiation composition, its preparation method, and its application. This ophthalmic composition includes SOD, trehalose, and tetrahydropyrimidine or an ophthalmologically acceptable tetrahydropyrimidine derivative. The SOD constitutes 0.05%-2% by mass, the trehalose 0.1%-5% by mass, and the tetrahydropyrimidine or an ophthalmologically acceptable tetrahydropyrimidine derivative 0.1%-4% by mass. This ophthalmic composition, by combining SOD, trehalose, and tetrahydropyrimidine in a certain mass ratio, can stabilize the lacrimal glands, relieve dry eyes, protect the eyes from blue light radiation, scavenge free radicals in the eyes, and improve symptoms such as eye fatigue, redness, and inflammation caused by excessive eye strain. However, this composition contains quaternary ammonium salts with antibacterial and preservative properties. Long-term use may lead to corneal lesions such as corneal calcification in some patients, causing premature cataracts and presbyopia, and also increasing myopia.

[0006] Therefore, there is an urgent need in the art to provide an eye-use composition that provides long-lasting moisturization, better hydration, longer duration of action, and prevention of cataracts. Summary of the Invention

[0007] This invention addresses the problems existing in the prior art by providing a multifunctional composition containing ergothioneine, its preparation method, and its application in eye products.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A multifunctional composition containing ergothioneine, comprising ergothioneine, tromethamine, and sodium hyaluronate in a mass ratio of 0.5-10:1-5:0.5-3.

[0009] Preferably, the mass ratio of ergothioneine, tromethamine, and sodium hyaluronate is 5-10:1-3:0.5-2.

[0010] More preferably, the mass ratio of ergothioneine, tromethamine, and sodium hyaluronate is 5:1.5:1.

[0011] Preferably, the multifunctional composition further includes a polyol, an osmotic pressure regulator, and water for injection.

[0012] Preferably, the polyol is selected from at least one of diols, tetraols, pentaols, and hexaols.

[0013] More preferably, the polyol is selected from at least one of glycerol, propylene glycol, sorbitol, erythritol and xylitol.

[0014] More preferably, the polyol is glycerol, erythritol, or xylitol.

[0015] Preferably, the osmotic pressure regulator is sodium chloride.

[0016] Preferably, the multifunctional composition comprises, by weight percentage, 0.05%-1% ergothioneine, 0.1%-0.5% tromethorphan, 0.05%-0.3% sodium hyaluronate, 0.1%-2% polyol, 0.15%-0.85% osmotic pressure regulator, and water for injection to bring the total to 100%.

[0017] More preferably, the multifunctional composition comprises, by weight percentage, 0.5%-1% ergothioneine, 0.1%-0.3% tromethamine, 0.05%-0.2% sodium hyaluronate, 0.2%-0.3% polyol, 0.5%-0.85% osmotic pressure regulator, and water for injection to bring the total to 100%.

[0018] More preferably, the multifunctional composition comprises, by weight percentage, 0.5% ergothioneine, 0.15% tromethamine, 0.1% sodium hyaluronate, 0.25% polyol, 0.80% osmotic pressure regulator, and water for injection to bring the total to 100%.

[0019] The present invention also provides a method for preparing the above-mentioned multifunctional composition, comprising the following steps: (1) First, mix sodium hyaluronate, polyol and water for injection to obtain mixture A; (2) Then mix ergothioneine and tromethamine to obtain mixture B; (3) Finally, mix mixture A and mixture B evenly, and then add osmotic pressure regulator to adjust the osmotic pressure to 260-340 mOsm / (Kg·H2O), and the mixture is obtained.

[0020] Preferably, the mixing in step (1) needs to be heated to 50-60°C and kept at that temperature for 10-20 minutes.

[0021] Preferably, the mixing temperature in step (2) is 20-40℃, and the pH is adjusted to 6.5-7.0 after mixing.

[0022] Preferably, the mixing temperature in step (3) is 20-50°C.

[0023] The present invention also provides the application of the above-described multifunctional composition or the multifunctional composition prepared by the above-described preparation method in eye products.

[0024] Preferably, the eye product is a medicine for the prevention of cataracts.

[0025] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention provides a multifunctional composition containing ergothioneine, wherein the tromethamine in the composition can protect the stability of ergothioneine under light and high temperature conditions; ergothioneine can also protect the activity of large molecular sodium hyaluronate. By utilizing the synergistic combination of ergothioneine, sodium hyaluronate and tromethamine, the moisturizing effect and eye protection effect of the composition can be significantly improved.

[0026] (2) The multifunctional composition containing ergothioneine provided by the present invention, through specific components and proportions, has the effect of preventing cataracts, and has long-lasting eye moisturizing effect and good stability. Detailed Implementation

[0027] It is worth noting that the raw materials used in this invention are all commercially available products.

[0028] Example 1 A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.5% ergothioneine, 0.15% tromethamine, 0.1% sodium hyaluronate (molecular weight 1000 kDa), 0.25% glycerol (i.e., polyol), 0.8% sodium chloride, and water for injection to bring the total to 100%.

[0029] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, glycerin and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 30°C, and then adjust the pH to 7.00 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0030] Example 2 A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.8% ergothioneine, 0.3% tromethamine, 0.2% sodium hyaluronate (molecular weight 1000 kDa), 0.2% mannitol, 0.4% sorbitol, 0.6% sodium chloride, and water for injection to bring the total to 100%.

[0031] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, mannitol, sorbitol and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 30°C, and then adjust the pH to 6.50 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 45°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0032] Example 3 A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.05% ergothioneine, 0.1% tromethorphan, 0.3% sodium hyaluronate (molecular weight 1000 kDa), 2% xylitol, 0.5% sodium chloride, and water for injection to bring the total to 100%.

[0033] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, xylitol and water for injection at 50°C and keep warm for 20 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 40°C, and then adjust the pH to 6.80 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 50°C, and then add potassium chloride to adjust the osmotic pressure to 320 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0034] Example 4 A multifunctional composition containing ergothioneine, comprising, by weight percentage: 1% ergothioneine, 0.5% tromethorphanol, 0.05% sodium hyaluronate (molecular weight 1000 kDa), 0.1% erythritol, 0.75% sodium chloride, and water for injection to bring the total to 100%.

[0035] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, erythritol and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 20°C, and then adjust the pH to 7.00 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 20°C, and then add potassium chloride to adjust the osmotic pressure to 280 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0036] Comparative Example 1 Compared to Example 1, the only difference is that the multifunctional composition does not contain tromethamine and sodium hyaluronate.

[0037] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.75% ergothioneine, 0.25% glycerin, 0.84% ​​sodium chloride, and water for injection to bring the total to 100%.

[0038] The preparation method of this multifunctional composition is as follows: (1) First, mix glycerol and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Finally, mix mixture A and ergot thiosulfate at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0039] Comparative Example 2 Compared to Example 1, the only difference is that the multifunctional composition does not contain ergothioneine.

[0040] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.45% tromethamine, 0.3% sodium hyaluronate (molecular weight 1000 kDa), 0.25% glycerin, 0.83% sodium chloride, and water for injection to bring the total to 100%.

[0041] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, glycerin and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Finally, mix mixture A and tromethorphan at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0042] Comparative Example 3 Compared to Example 1, the only difference is that the multifunctional composition does not contain tromethamine.

[0043] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.5% ergothioneine, 0.25% sodium hyaluronate (molecular weight 1000 kDa), 0.25% glycerin, 0.8% sodium chloride, and water for injection to bring the total to 100%.

[0044] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, glycerin and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Finally, mix mixture A and ergothioneine at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0045] Comparative Example 4 Compared to Example 1, the only difference is that the multifunctional composition does not contain sodium hyaluronate.

[0046] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.5% ergothioneine, 0.25% tromethorphan, 0.25% glycerin, 0.8% sodium chloride, and water for injection to bring the total to 100%.

[0047] The preparation method of this multifunctional composition is as follows: (1) First, mix glycerol and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 30°C, and then adjust the pH to 7.00 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0048] Comparative Example 5 Compared to Example 1, the only difference is that glycerol is replaced with mannitol.

[0049] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.5% ergothioneine, 0.15% tromethamine, 0.1% sodium hyaluronate (molecular weight 1000 kDa), 0.25% mannitol, 0.82% sodium chloride, and water for injection to bring the total to 100%.

[0050] The preparation method of this multifunctional composition is as follows: (1) First, mix sodium hyaluronate, mannitol and water for injection at 60°C and keep warm for 15 min to obtain mixture A; (2) Then mix ergothioneine and tromethamine at 30°C, and then adjust the pH to 7.00 to obtain mixture B; (3) Finally, mix mixture A and mixture B at 50°C, and then add sodium chloride to adjust the osmotic pressure to 300 mOsm / (Kg·H2O) to obtain a multi-functional composition.

[0051] Comparative Example 6 Compared with Example 1, the only difference is the component ratio.

[0052] A multifunctional composition containing ergothioneine, comprising, by weight percentage: 0.5% ergothioneine, 0.8% tromethorphan, 0.4% sodium hyaluronate (molecular weight 1000 kDa), 0.3% glycerin, 0.8% sodium chloride, and water for injection to bring the total to 100%.

[0053] The preparation method of this multifunctional composition is the same as that in Example 1.

[0054] Test Example 1—High Temperature and Light Stability High temperature and light stability tests were conducted on the multifunctional compositions prepared in Examples 1-4 and Comparative Examples 1, 3, 4 and 5, respectively.

[0055] The high-temperature stability test method is as follows: The multi-functional composition was placed at a high temperature of 40±2℃ / RH 75±5% for 6 months. The results were examined at the 6th month and compared with those at day 0. The appearance of the multi-functional composition under different formulations was examined. The ergothionein content was determined by high performance liquid chromatography (chromatographic conditions: column: Agilent ZORBAX Eclipse XDB-C18 column (4.6×250 mm, 5 μm); column temperature: 25℃; mobile phase: 60% acetonitrile; detection wavelength: 254 nm; injection volume: 5 μL; flow rate: 0.65 mL / min; run time: 20 min; blank solution: 60% acetonitrile).

[0056] The light stability test method is as follows: The multifunctional composition is placed under light exposure of 4500±500 lx for 15 days, and the relative retention rate of the formulation under extremely destructive conditions is observed. This method is only for rapid screening of formulations; in practical applications, a light-proof container must be used.

[0057] The high-temperature stability data are shown in Table 1, and the light stability data are shown in Table 2. As can be seen from Tables 1 and 2, the ergothioneine in Examples 1-4 has a retention rate of >97% at 40℃ and 6 months. Under extreme destructive conditions, the ergothioneine retention rate is >60%. In Comparative Examples 1 and 3, the degradation of ergothioneine is accelerated due to the lack of tromethamine. Sodium hyaluronate in Comparative Example 4 can improve the stability of the formulation.

[0058] Table 1 High Temperature Stability Test Data

[0059] Table 2. Light stability test data

[0060] Test Example 2—Protecting the activity of macromolecular HA Sodium hyaluronate (HA) is a key ingredient in ophthalmic products. As a core component of artificial tears, it utilizes its powerful moisturizing and lubricating abilities to relieve dry eye discomfort. However, HA is susceptible to molecular weight degradation under the influence of free radicals, leading to a decrease in the overall system viscosity and a decline in its moisturizing activity, thus diminishing its eye-protecting function. Ergothioneine, as a strong antioxidant, can effectively maintain the molecular weight of HA and the viscosity stability of the system, thereby preserving the activity of HA.

[0061] By accelerating experimental design, using Cu 2+The protective effects of HA dosage and ergothioneine dosage at different dosages were tested by comparing Examples 1-3 and Comparative Example 2, and the oxidative degradation of HA by ascorbic acid. The experimental methods are as follows: A 16 mM (2.818 g / L) ascorbic acid solution and a 0.1 mM (0.017 g / L) copper chloride solution were prepared simultaneously with the multifunctional compositions of Examples 1-3 and Comparative Example 2. After mixing, HA with a molecular weight of 1000 kDa was added. After stirring for 5 hours, the viscosity change of the system was tested, and the viscosity retention rate of the system was calculated. The calculation formula is as follows: System viscosity retention rate (%) = viscosity after 5 h ÷ initial viscosity × 100%.

[0062] The experimental results are shown in Table 3 below.

[0063] Table 3 Viscosity Change

[0064] Test Example 3—Synergistic Moisturizing Composition Soft contact lenses were immersed in the compositions prepared in Example 1 and Comparative Examples 1-4 for 24 hours, with the solution changed three times during the process to ensure that the lenses absorbed the composition. The lenses were then removed from the composition and placed on a dry, clean, absorbent cloth free of lint. The cloth was folded over the lenses, and the surface moisture was gently absorbed with fingers. The lenses were then placed in a constant-weight glass weighing dish, and the initial weight and the time immediately after drying were recorded. The weighing dish was placed in a desiccator with 44% humidity (the operating environment humidity was controlled at 44±5%). After 3 hours, the lenses were removed and weighed, and the moisture retention rate was calculated using the following formula.

[0065]

[0066] In the formula: W0 is the initial weight of the lens, g; Wn is the weight of the lens after 3 hours, g.

[0067] The data results are shown in Table 4.

[0068] Table 4. Results of Moisturizing Rate Test

[0069] Test Example 4—Pharmacodynamic Test Pharmacodynamic tests for the prevention of cataracts were conducted on the multifunctional compositions prepared in Example 1 and Comparative Examples 1-6, respectively. The experimental methods are as follows: One hundred Wistar rats, aged 12 days and weighing 18-20 g, were used, with half being male and half female. The rats were randomly divided into 10 groups (model group, positive drug group, Example 1 group, and comparative groups 1-6). Except for the blank control group, the rats in the other groups were injected intraperitoneally with 0.01 L / kg of 0.05% sodium selenite solution to induce the model.

[0070] Three days before modeling, after the suckling mice opened their eyes, they were given the corresponding drug preparations. The positive control group was given 0.5% pyrenoxine sodium eye drops, while the Example 1 group and Comparative Examples 1-6 groups were given the corresponding combinations. Administration continued until the 7th day after modeling, twice daily at a dose of 15 g / (kg·d). The blank control group and the model group were given an equal volume of physiological saline. One day before and one day after the start of administration, the mice were dilated with 1% atropine sulfate solution. In a dark room, the opacity of the lens in both eyes was observed using a handheld slit lamp, and the lens opacity was compared.

[0071] For evaluation criteria, please refer to: "Effects of Zinc Supplementation on LDH and CuZn-SOD in Blood and Lens of Cataract Rats", where "-" indicates a transparent lens; "+" indicates a cloudy lens nucleus but a clear cortex; and "*" indicates a cloudy lens nucleus and cortex.

[0072] The data results are shown in Table 5. As can be seen from Table 5, the multifunctional composition of Example 1 of the present invention has a very significant preventive effect on cataracts.

[0073] Table 5 Test Results

[0074] Test Example 5—Evaluation of Usage Effectiveness The efficacy of the multi-functional compositions prepared in Examples 1-4 and Comparative Examples 1-6 was evaluated using the following methods: a single-center, randomized, open-label, parallel, and self-controlled study was conducted. Inclusion criteria included: subjective symptoms such as dry eyes, foreign body sensation, burning sensation, fatigue, discomfort, redness, and fluctuating vision; a score of ≥7 on the Chinese Dry Eye Questionnaire; and signed informed consent. A total of 50 subjects were enrolled and randomly divided into 10 groups (5 subjects per group). Each group used 0.3 mL of the multi-functional compositions prepared in Examples 1-4 and Comparative Examples 1-6 as eye drops once daily, morning and evening, for 14 consecutive days.

[0075] Qualified subjects underwent two visits during the enrollment period (Day 0) and the exit period (Day 14). At each visit, the Dry Eye Questionnaire, the Ocular Surface Disease Index (OSDI) score, and the general survey version of the Visual Fatigue Scale were completed. Fluorescein staining of tear film breakup time and corneal staining were also performed. Changes in the scores of the Dry Eye Questionnaire, the Ocular Surface Disease Index (OSDI) score, and the general survey version of the Visual Fatigue Scale were compared across the different visit periods. The rate of increase in fluorescein-stained tear film breakup time compared to Day 0 and the incidence of adverse reactions during the trial period were calculated. The results are shown in Table 6.

[0076] Table 6. Results of Therapeutic Effect Evaluation

[0077] Note: Compared to Day0, * indicates p < 0.05, and *** indicates p < 0.001.

[0078] This study included 50 subjects. Except for control group 6, the remaining groups experienced no adverse events after use, and the adverse event rate was 0%. No subjects withdrew from the study due to adverse events. The comprehensive safety analysis results indicate that the multi-functional composition containing ergothioneine of this invention has good overall safety.

[0079] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A multifunctional composition containing ergothioneine, characterized in that, The multifunctional composition comprises, by weight percentage: 0.05%-1% ergothioneine, 0.1%-0.5% tromethamine, 0.05%-0.3% sodium hyaluronate, 0.1%-2% polyol, 0.15%-0.85% osmotic pressure regulator, and water for injection to bring the total to 100%. The mass ratio of ergothioneine, tromethamine, and sodium hyaluronate is 5-10:1-3:0.5-2; The polyol is glycerol or a combination of mannitol and sorbitol.

2. The multifunctional composition according to claim 1, characterized in that, The osmotic pressure regulator is sodium chloride.

3. A method for preparing a multifunctional composition as described in claim 1 or 2, characterized in that, Includes the following steps: (1) First, mix sodium hyaluronate, polyol and water for injection to obtain mixture A; (2) Then mix ergothioneine and tromethamine to obtain mixture B; (3) Finally, mix mixture A and mixture B evenly, and then add osmotic pressure regulator to adjust the osmotic pressure to 260-340 mOsm / (Kg·H2O), and the mixture is obtained.

4. The preparation method according to claim 3, characterized in that, The mixing in step (1) needs to be heated to 50-60℃ and kept warm for 10-20 min; the mixing temperature in step (2) is 20-40℃, and the pH is adjusted to 6.5-7.8 after mixing; the mixing temperature in step (3) is 20-50℃.

5. The use of a multifunctional composition as described in claim 1 or 2, or a multifunctional composition prepared by the preparation method as described in claim 3 or 4, in the preparation of eye products.

6. The application according to claim 5, characterized in that, The eye product in question is a medication for the prevention of cataracts.

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