A ternary eutectic solvent containing bosonicine, its preparation method and application

Abstract

This invention relates to the field of eutectic solvent technology, specifically to a ternary eutectic solvent containing PHOXIN, its preparation method, and its applications. In this ternary eutectic solvent containing PHOXIN, the invention proposes for the first time the preparation of PHOXIN into a ternary eutectic solvent. This eutectic solvent can promote the skin penetration of PHOXIN and improve its bioavailability. Simultaneously, this ternary eutectic solvent can also be used as a solvent to dissolve other active ingredients, thereby expanding its applications in skincare products, cosmetics, and topical pharmaceuticals.

Description

Technical Field

[0001] This invention relates to the field of eutectic solvent technology, and in particular to a ternary eutectic solvent containing bosonicine, its preparation method, and its application. Background Technology

[0002] Ionic liquids (ILs) were discovered by Paul Walden in 1914 while exploring alternatives to nitroglycerin used in explosives. Ionic liquids are defined as molten salts primarily composed of a mixture of organic cations and organic or inorganic anions. The melting point of ionic liquids is approximately 100°C, lower than the melting points of their individual components. Deep eutectic solvents (DES) are organic solvents with thermodynamic properties similar to ILs. DES consist of two main components: hydrogen bond donors (HBDs), such as acids, amines, or alcohols, and hydrogen bond acceptors (HBAs), such as tetraalkylammonium, quaternary ammonium salts, or phosphonium salts. DES can be prepared by heating a mixture of HBAs and HBDs until a homogeneous, transparent solution is formed, or by dissolving the individual components in a solvent and then evaporating the solvent. The melting point of DES is significantly lower than that of its individual components. Eutectic solvents are commonly used in the extraction of active plant components and in research on enhancing the penetration of topical drugs. For example, Chinese invention patent CN114177211A discloses a method for preparing a eutectic solvent-traditional Chinese medicine extract mixture system. This method involves mixing the target traditional Chinese medicine extract with a hydrogen bond donor and a hydrogen bond acceptor, followed by heating to obtain the eutectic solvent-traditional Chinese medicine extract mixture system. Using this mixture for transdermal delivery can improve the penetration rate of complex component clusters in traditional Chinese medicine. Chinese invention patent application CN112190540A discloses a type of nasal penetration enhancer and its application. This penetration enhancer is a low-temperature molten salt that promotes nasal absorption. It is formed by the reaction of an organic acid with at least one organic base and / or its salt to form an ionic liquid or eutectic solvent. This penetration enhancer can dissolve and absorb poorly soluble drugs and biomolecular drugs.

[0003] Pro-Xylane, full name (2S,3R,4S,5R)-2-(2-hydroxypropyl)tetrahydro-2H-pyran-3,4,5-triol, or hydroxypropyltetrahydropyrantriol for short, is a bioactive substance derived from natural xylose. It can activate the synthesis of mucopolysaccharides, promote the production of hyaluronic acid, improve the adhesion between the dermis and epidermis, promote the synthesis of collagen VII and collagen IV, regenerate damaged tissue, maintain the elasticity of the dermis, and prevent skin aging. Pro-Xylane is a water-soluble ingredient. Since the stratum corneum of human skin is lipophilic, conventional Pro-Xylane has difficulty penetrating the stratum corneum to reach the subcutaneous tissue and exert its effects. In the cosmetics industry, Pro-Xylane products are usually based on high concentrations to effectively express their efficacy. However, a large amount of Pro-Xylane is not absorbed by the skin and remains on the skin surface, where it is washed away or metabolized, resulting in low utilization and high prices for related products. Whether using eutectic solvents can effectively promote the transdermal absorption of bosonicine is a topic that has not been studied in the current technology. Summary of the Invention

[0004] To address the aforementioned technical problems, the present invention aims to provide a ternary eutectic solvent containing BOXIN, its preparation method, and its application. In this ternary eutectic solvent containing BOXIN, the present invention, for the first time, proposes to prepare BOXIN into a ternary eutectic solvent. This eutectic solvent can promote the skin penetration of BOXIN and improve its bioavailability. Simultaneously, this ternary eutectic solvent can also be used as a solvent to dissolve other active ingredients, thereby expanding its application in skincare products, cosmetics, and topical pharmaceuticals.

[0005] To achieve the above-mentioned technical effects, the present invention adopts the following technical solution:

[0006] In a first aspect, this aspect provides a ternary eutectic solvent containing a bosine, the ternary eutectic solvent including a bosine, and further including a hydrogen bond acceptor and a hydrogen bond donor.

[0007] Preferably, the molar ratio of the bosonic, hydrogen bond acceptor, and hydrogen bond donor is 1-3:1-2:1-6.

[0008] Most preferably, the molar ratio of the bosonic, hydrogen bond acceptor, and hydrogen bond donor is 1:2:6.

[0009] Furthermore, the hydrogen bond acceptor is any one or more of betaine, L-carnitine, and matrine.

[0010] Furthermore, the hydrogen bond donor is any one or more of glycerol, propylene glycol, xylitol, erythritol, and butylene glycol.

[0011] Preferably, the hydrogen bond acceptor is betaine and the hydrogen bond donor is xylitol.

[0012] Secondly, the present invention provides a method for preparing a ternary eutectic solvent containing Bosein, comprising the steps of:

[0013] Take appropriate molar ratios of Bosein, hydrogen bond donor, and hydrogen bond acceptor substances, and stir and melt them at a set temperature until they become a clear and transparent liquid, thus obtaining a ternary eutectic solvent containing Bosein.

[0014] Preferably, the set temperature is 40–110°C.

[0015] Thirdly, the present invention also provides the application of a ternary eutectic solvent containing bosine in any aspect of the preparation of skin care products, cosmetics or topical medicines.

[0016] Preferably, the skin care product or cosmetic product contains at least ergothioneine.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] First, this invention provides a ternary eutectic solvent containing Bosein, which solves the problem of poor transdermal performance of Bosein and exhibits good biocompatibility and high stability. Second, the ternary eutectic solvent provided by this invention, by compounding Bosein with betaine and xylitol, can further enhance the expression of Bosein and other active ingredients dissolved in the ternary eutectic solvent compared to traditional binary eutectic solvents or other similar ternary eutectic solvents. Third, the preparation method of this ternary eutectic solvent containing Bosein is simple and feasible, easy to control, has high production efficiency, and can be mass-produced, playing a significant role in the fields of cosmetics, skin care products, and topical pharmaceuticals. Attached Figure Description

[0019] Figure 1 An appearance photograph of the ternary eutectic solvent containing bosonicine prepared in Example 1 of the present invention, which is provided as Test Example 1 of the present invention;

[0020] Figure 2 The DSC spectrum of the bosonic acid monomer provided in Experimental Example 1 of this invention;

[0021] Figure 3 The DSC spectrum of the betaine monomer provided in Experimental Example 1 of this invention;

[0022] Figure 4 The DSC spectrum of the xylitol monomer provided in Experimental Example 1 of this invention;

[0023] Figure 5 The DSC spectrum of the betaine-xylitol binary DES provided in Experimental Example 1 of this invention;

[0024] Figure 6The DSC spectrum of the Bosein-betaine-xylitol ternary DES provided in Experimental Example 1 of this invention;

[0025] Figure 7 The results of the 1H NMR spectrum detection of the Bosein monomer provided in Experimental Example 1 of this invention;

[0026] Figure 8 The results of the 1H NMR spectrum detection of the betaine monomer provided in Experimental Example 1 of this invention;

[0027] Figure 9 The results of the 1H NMR spectrum detection of xylitol monomer provided in Experimental Example 1 of this invention;

[0028] Figure 10 The results of the 1H NMR spectrum detection of the betaine-xylitol binary DES provided in Experimental Example 1 of this invention;

[0029] Figure 11 The results of the 1H NMR spectrum of the Bosein-betaine-xylitol ternary DES provided in Experimental Example 1 of this invention;

[0030] Figure 12 The results of the detection of BOXER skin retention amount in Example 1, Comparative Example 1, and Comparative Example 2 of the present invention;

[0031] Figure 13 The results of the detection of the amount of BOXER in the skin in a series of BOXER ternary DES of this invention;

[0032] Figure 14 The results of ergothionein skin retention in samples 1 and 2 of this invention;

[0033] Figure 15 The results of the anti-wrinkle efficacy test for human body provided by Sample 1 and Sample 2 of this invention. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The specific embodiments listed in the present invention are only examples of the present invention, and the present invention is not limited to the specific embodiments described below.

[0035] For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of this invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of this invention should be covered within its scope. Unless otherwise specified in the embodiments, conventional conditions or conditions recommended by the manufacturer should be followed. All reagents or instruments without a specified manufacturer are commercially available conventional products.

[0036] To better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art will understand that the present invention can be practiced without certain specific details. In other embodiments, methods, means, apparatus, and steps well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Unless otherwise specified, all units used in this specification are International Standard Units (SI), and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors unavoidable in industrial production.

[0038] The present invention will now be described in detail through specific embodiments. It should be noted that these embodiments are only for further illustration and should not be construed as limiting the scope of protection of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention based on the above description. All raw materials and reagents used in this invention are commercially available products.

[0039] Example 1

[0040] This embodiment provides a ternary eutectic solvent and its preparation method, specifically:

[0041] A ternary eutectic solvent containing Bosein was prepared by heating a mixture of Bosein, betaine, and xylitol in a ratio of 1 mol: 2 mol: 6 mol at 110 °C until melted and holding at that temperature for 1 hour.

[0042] Example 2

[0043] This embodiment provides a ternary eutectic solvent and its preparation method, specifically:

[0044] A ternary eutectic solvent containing Bosein was prepared by heating a mixture of Bosein, betaine, and xylitol in a ratio of 3 mol: 2 mol: 6 mol at 100 °C until melted and holding at that temperature for 1 hour.

[0045] Example 3

[0046] This embodiment provides a ternary eutectic solvent and its preparation method, specifically:

[0047] A ternary eutectic solvent containing Bosein was prepared by heating a mixture of Bosein, betaine, and xylitol in a ratio of 3 mol: 1 mol: 3 mol at 100°C until melted and holding at that temperature for 2 hours.

[0048] Example 4

[0049] This embodiment provides a ternary eutectic solvent and its preparation method, specifically:

[0050] A ternary eutectic solvent containing Bosein was prepared by heating a mixture of Bosein, betaine, and xylitol in a ratio of 1 mol: 1 mol: 1 mol at 100°C until melted and holding at that temperature for 1 hour.

[0051] Examples 5-18

[0052] Each contains 1 mole of Bosein, 2 moles of HBA, and 6 moles of HBD. The composition of HBA and HBD and the preparation temperature are shown in the table below. The preparation method is in accordance with Example 1.

[0053] Table 1. Composition and preparation temperature of Examples 5-18

[0054] Example 5 L-carnitine Xylitol 100 Example 6 matrine Xylitol 100 Example 7 betaine glycerin 60 Example 8 L-carnitine glycerin 60 Example 9 matrine glycerin 50 Example 10 betaine Propylene glycol 40 Example 11 L-carnitine Propylene glycol 60 Example 12 matrine Propylene glycol 50 Example 13 betaine Erythritol 100 Example 14 L-carnitine Erythritol 90 Example 15 matrine Erythritol 90 Example 16 betaine Butylene glycol 40 Example 17 L-carnitine Butylene glycol 50 Example 18 matrine Butylene glycol 40

[0055] Comparative Example 1

[0056] This embodiment provides a ternary eutectic solvent and its preparation method, which differs from Example 1 in that:

[0057] In this embodiment, Bosein, betaine, and xylitol were prepared in a molar ratio of 1:2:6, and the preparation method is as follows:

[0058] Weigh out Bosein, betaine, and xylitol according to the molar ratio, add them together to water, stir well, and prepare a mixed aqueous solution containing Bosein to obtain reference standard 1.

[0059] Comparative Example 2

[0060] This embodiment provides a ternary eutectic solvent and its preparation method, which differs from Example 1 in that:

[0061] In this embodiment, Bosein, betaine, and xylitol were prepared in a molar ratio of 1:2:6, and the preparation method is as follows:

[0062] First, betaine-xylitol binary DES was prepared, and the preparation method is the same as in Example 1;

[0063] Then, weigh out the bosine according to the molar ratio and add it to the aforementioned betaine-xylitol binary DES. Mix it evenly at room temperature to obtain reference standard 2.

[0064] Comparative Example 3

[0065] This embodiment provides a binary eutectic solvent and its preparation method, which differs from Example 1 in that:

[0066] In this embodiment, betaine and xylitol are prepared in a molar ratio of 2:6, and the preparation method is as follows:

[0067] A binary eutectic solvent containing betaine and xylitol was prepared by heating the betaine and xylitol mixture at a ratio of 2 mol: 6 mol at 100°C until it melted and holding it at that temperature for 1 hour.

[0068] Experimental Example 1

[0069] 1.1 Appearance Inspection

[0070] The ternary eutectic solvent prepared in Example 1 was subjected to visual inspection, such as... Figure 1 As shown.

[0071] 1.2 DSC Differential Scanning Calorimeter for Melting Point Determination

[0072] For the bosonic monomer, the DSC spectrum of bosonic was obtained by heating from 0℃ to 150℃ at a rate of 10℃ / min, as shown below. Figure 2 As shown;

[0073] For betaine monomers, DSC spectra of betaine were obtained at temperatures ranging from 200℃ to 350℃ with a heating rate of 10℃ / min, as shown below. Figure 3 As shown;

[0074] For xylitol monomers, DSC spectra of xylitol were obtained by heating from 0℃ to 150℃ at a rate of 10℃ / min, as shown below. Figure 4 As shown;

[0075] The betaine-xylitol binary DES prepared in Comparative Example 3 was subjected to DSC spectroscopy at a temperature ranging from -50℃ to 100℃ at a heating rate of 10℃ / min. The results are shown below. Figure 5 As shown;

[0076] The ternary eutectic solvent of Bosein-betaine-xylitol obtained in Example 1 was used at a temperature ranging from -50°C to 100°C at a heating rate of 10°C / min to obtain the DSC spectrum of the ternary eutectic solvent, as shown below. Figure 6 As shown.

[0077] The above experimental results show that the ternary eutectic solvent containing bosonicine prepared in Example 1 is homogeneous and has a significantly reduced melting point.

[0078] 1.3 Detection by 1H NMR spectroscopy

[0079] The monomers of each component, the betaine-xylitol binary DES prepared in Comparative Example 3, and the ternary eutectic solvent containing Bosein prepared in Example 1 were analyzed by 1H NMR spectroscopy. The detection conditions were as follows:

[0080] Nuclear magnetic resonance spectrometer: Bruker, Switzerland, AVANCE NEO 400;

[0081] Frequency: 400MHz;

[0082] Injection volume: 5 mg;

[0083] Solvent: Deuterated DMSO;

[0084] The detection method is as follows:

[0085] Five mg each of Bosein monomer, betaine monomer, xylitol monomer, the betaine-xylitol binary DES prepared in Comparative Example 3, and the Bosein-betaine-xylitol ternary eutectic solvent obtained in Example 1 were dissolved in 0.6 mL of deuterated DMSO reagent and detected by proton nuclear magnetic resonance spectroscopy.

[0086] The results are as follows: Figures 7-11 As shown above, the results indicate that the hydrogen elution positions of the ternary eutectic solvent prepared in Example 1 are shifted to a certain extent compared with the individual monomers and the betaine-xylitol binary DES, proving that the ternary eutectic solvent is a new eutectic substance rather than a simple physical mixture.

[0087] 1.4 Study on the amount of Brønsted sorbitan retained in the skin

[0088] The transdermal performance of the ternary eutectic solvent prepared in Example 1 was tested using bosonicine, under the following conditions:

[0089] Liquid Chromatograph: Agilent 1260 Infinity II

[0090] Chromatographic conditions: Inertsil HPLC column (4.6 mm × 250 mm, 5 μm); mobile phase: water: acetonitrile = 60:40; flow rate: 0.5 mL / min; column temperature: 30 °C; injection volume: 2 μL.

[0091] The detection method is as follows:

[0092] (1) Establishment of the Bosonic standard curve:

[0093] A suitable amount of Bosein was weighed into a brown volumetric flask, dissolved in pure water, and diluted to volume to prepare solutions with concentration gradients of 1, 2, 5, 10, 20, 50, 100, 200, 500, and 1000 μg / mL. These solutions were then detected by HPLC. The results showed that the linear fitting equation was y = 13.86x + 1.7621 (R² = 1), exhibiting good linearity in the range of 1–1000 μg / mL.

[0094] (2) Study on skin retention

[0095] Before the experiment, a pigskin model with a thickness of 300±50 μm was prepared using a skin grafting scalpel. This model was then cut into small circular pieces the size of the receiving pool and placed in an equal volume of physiological saline. Phosphate-buffered saline (pH=7.4) was used as the receiving medium. During the experiment, the skin model was fixed between the release pool and the receiving pool, with the stratum corneum side facing the release pool and the dermis side facing the receiving pool, ensuring close contact between the skin and the receiving solution, and preventing air bubbles from forming. Subsequently, a certain amount of the samples from Example 1, Comparative Example 1, and Comparative Example 2 were added to the skin surface, respectively. The temperature of the receiving pool was maintained at 37±0.5℃, and a magnetic magnet was placed inside the receiving pool, rotating at 300 rpm throughout the experiment. Each group had three replicates.

[0096] Six hours after the in vitro percutaneous treatment, the skin was removed, and any residual solution on the surface was rinsed off with pure water. The skin from the transdermal site was then minced, homogenized with 1.5 ml of PBS, and sonicated for 1 hour to extract Brønsted skin. The homogenate was transferred to a centrifuge tube, vortexed, and centrifuged at 12000 rpm for 10 min. The residue was extracted again with 1 ml of PBS. The supernatants from both extractions were combined, mixed, and the supernatant was filtered through a 0.22 μm filter membrane. The sample was analyzed using the high-performance liquid chromatography (HPLC) method described above to determine the amount of Brønsted skin retained per unit area. The results are recorded in Table 2. Figure 12 .

[0097] Table 2. Statistical results of the average amount of BPosein retained per unit area of ​​skin.

[0098]

[0099] The results showed that the amount of Bosein retained on the skin in the ternary eutectic solvent containing Bosein was 2.78 times that of Comparative Example 1 and 1.45 times that of Comparative Example 2. The ternary eutectic solvent in this invention has a penetration-enhancing effect.

[0100] 1.5 Comparative Study on the Retention Amount of Pro-Xylane in Skin of Various Pro-Xylane Trivalent DES

[0101] Various types of ternary DES containing bosoxane were prepared according to Examples 1 and 5-18. Based on the skin retention study method in section 1.4(2), a comparative study of the skin retention of bosoxane DES was conducted. The experimental results are shown in Table 3 and... Figure 13 .

[0102] Table 3. Statistical results of the average cumulative skin retention of PVXin in various ternary DES containing PVXin.

[0103]

[0104] The results showed that the skin retention of the ternary eutectic solvent of bosine-xylitol-betaine was higher than that of other ternary eutectic solvents prepared by HBD and HBA.

[0105] Experimental Example 2

[0106] This experiment aims to investigate the effect of the ternary eutectic solvent of bosine-xylitol-betaine prepared in Example 1 on the skin penetration rate of ergothioneine. The experimental method is as follows:

[0107] 2.1 Establishment of the ergothioneine standard curve

[0108] (1) The liquid phase analysis method is as follows:

[0109] Liquid Chromatograph: Agilent 1260 Infinity II

[0110] Chromatographic conditions: Hypersil ODS C18 column (250 mm × 4.6 mm, 5 μm); mobile phase: acetonitrile-water (3:97); flow rate: 1.0 mL / min; column temperature: 30 ℃; detection wavelength: 254 nm; injection volume: 20 μL.

[0111] (2) Ergothioneine standard curve:

[0112] An appropriate amount of ergothioneine was weighed into a brown volumetric flask, dissolved in pure water, and diluted to volume to prepare solutions with concentration gradients of 1, 2, 5, 10, 20, 50, 100, 200, 500, and 1000 μg / mL. These solutions were then detected by HPLC. The results showed that the linear fitting equation was y = 7.0563x + 1.1693 (R² = 0.9999), exhibiting good linearity in the range of 1–1000 μg / mL.

[0113] 2.2 Preparation of two groups of samples before the experiment

[0114] Sample 1: Ergothioneine and the eutectic solvent of Example 1 were mixed at a molar ratio of ergothioneine to Boseine of 1:2 to prepare a Boseine-betaine-xylitol ternary DES solution containing ergothioneine.

[0115] Sample 2: Ergothioneine and the solvent of Comparative Example 1 were mixed at a molar ratio of ergothioneine to bosine of 1:2 to prepare a mixed aqueous solution containing ergothioneine, bosine, betaine and xylitol.

[0116] 2.3 Study on ergothioneine retention in the skin

[0117] Before the experiment, a pigskin model with a thickness of 300±50 μm was prepared using a skin grafting scalpel. This model was then cut into small circular pieces the size of the receiving pool and placed in an equal volume of physiological saline. Phosphate-buffered saline (pH=7.4) was used as the receiving medium. During the experiment, the skin model was fixed between the release pool and the receiving pool, with the stratum corneum side facing the release pool and the dermis side facing the receiving pool, ensuring close contact between the skin and the receiving solution, without any air bubbles. Subsequently, a certain amount of Sample 1 and Sample 2 were added to the skin surface, respectively. The temperature of the receiving pool was maintained at 37±0.5℃, and a magnetic magnet was placed inside the receiving pool, rotating at 300 rpm throughout the experiment. Each group had three replicates.

[0118] Six hours after the in vitro percutaneous treatment, the skin was removed, and any residual solution on the surface was rinsed off with pure water. The skin from the transdermal site was then minced, homogenized with 1.5 ml of PBS, and sonicated for 1 hour to extract ergothionein from the skin. The homogenate was transferred to a centrifuge tube, vortexed, and centrifuged at 12000 rpm for 10 min. The residue was extracted again with 1 ml of PBS, and the supernatants from both extractions were combined, mixed, and the supernatant was filtered through a 0.22 μm filter membrane. The sample was analyzed using the high-performance liquid chromatography (HPLC) method described above to determine the amount of ergothionein retained per unit area of ​​skin. The results are recorded in Table 4. Figure 14 .

[0119] Table 4. Statistical results of the average amount of ergothionein retained per unit area of ​​skin.

[0120] <![CDATA[Skin retention amount (μg / cm 2 )]]> 255.94 75.84 Standard deviation (SD) 35.13 16.32

[0121] The results showed that in the ternary eutectic solvent of ergothioneine and bosine, the skin retention of ergothioneine was 3.37 times that of the solution group, indicating that the ternary eutectic solvent in this invention has a significant effect on promoting the penetration of ergothioneine.

[0122] 2.4 Cellular Anti-wrinkle Efficacy Test

[0123] Select test groups: Sample 1 and Sample 2.

[0124] Cellular experiments were conducted using the following methods:

[0125] (1) Cell seeding: at an appropriate seeding density (8 x 10⁻⁶ cells / year). 4 Cells were seeded into 24 wells and incubated overnight in an incubator (37°C, 5% CO2).

[0126] (2) Experimental grouping: The experiment was set up with a blank control group, a positive control group and a sample group. The sample group was set with 3 concentration gradients.

[0127] (3) Solution preparation: Prepare working solutions of different concentrations of test substance according to the test concentration setting table.

[0128] (4) Drug administration: Drug administration was carried out when the cell deposition rate in the 24-well plate reached 40%-60%. 1 mL of cell culture medium was added to each well of the blank control group; 1 mL of culture medium containing 100 μg / mL vitamin C and 7 μg / mL vitamin E was added to each well of the positive control group; and 1 mL of culture medium containing the corresponding concentration of the test substance was added to each well of the sample group.

[0129] (5) Radiation: 24 hours after drug administration, the total dose received by the positive control group and the sample group was 9 J / cm². 2 The control group was placed under UVA radiation, while the blank control group was placed in the same environment (UVA radiation dose of 0 J / cm²). 2 ).

[0130] (6) Collect cell supernatant: After incubation for 24 hours, collect the cell culture supernatant in an EP tube and freeze it at -80°C.

[0131] (7) Collagen I detection: The content of Collagen I was detected and analyzed according to the instructions of the ELISA kit.

[0132] The test results are shown in Table 5.

[0133] Table 5. Statistical results of the mean relative content of Collagen I.

[0134]

[0135] The results show that, compared with sample 2, sample 1 showed a 24% increase in collagen content, indicating that the ternary eutectic solvent prepared by using BOXER-betaine-xylitol can enable BOXER and ergothionein to have a synergistic effect, which is more conducive to the anti-wrinkle effects of BOXER and ergothionein.

[0136] 2.5 Human Anti-wrinkle Efficacy Test

[0137] Sample 1 and Sample 2 were tested for their anti-wrinkle efficacy on human bodies.

[0138] Thirty-three people aged 30-60 were randomly selected, including 18 men and 15 women. Sample 1 was applied to the corners of their eyes every 2 days. The state of wrinkles at the corners of the eyes was recorded before and 28 days after application using a three-dimensional imaging system.

[0139] Thirty-three people aged 30-60 were randomly selected, including 14 men and 19 women. Sample 2 was applied to the corners of their eyes every 2 days. The wrinkles at the corners of their eyes were recorded before and 28 days after application using a three-dimensional imaging system.

[0140] Specific results are as follows Figure 15 As shown, Sample 1 showed significantly better improvement in crow's feet wrinkles than Sample 2. The results indicate that the addition of ergothioneine to the ternary eutectic solvent of Boseine-betaine-xylitol significantly improved the bioavailability of ergothioneine and Boseine compared to a physical mixture of ergothioneine, Boseine, betaine, and xylitol. This ternary eutectic solvent effectively enhanced the skin penetration of ergothioneine, promoted the bioavailability of Boseine and ergothioneine, and significantly improved their synergistic anti-wrinkle effect.

[0141] The above experimental results show that the ternary eutectic solvent containing serotonin provided by the present invention has great application value in the field of skin care products. In this system, not only can the penetration-enhancing effect of serotonin itself be achieved, but it can also further promote the transdermal absorption of ergothioneine active ingredient and synergistically enhance the effect with ergothioneine to achieve very significant anti-wrinkle effects on cells and the human body.

[0142] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention. Technical aspects, shapes, and structures not described in detail in this invention are all well-known technologies.

Claims

1. A ternary eutectic solvent containing bosonicine, characterized in that: It includes Bosein, hydrogen bond acceptor and hydrogen bond donor; the hydrogen bond acceptor is any one or more of betaine, L-carnitine and matrine; the hydrogen bond donor is any one or more of glycerol, propylene glycol, xylitol, erythritol and butylene glycol; the molar ratio of Bosein, hydrogen bond acceptor and hydrogen bond donor is 1-3:1-2:1-6.

2. The ternary eutectic solvent containing bosonicine as described in claim 1, characterized in that: The molar ratio of the bosonic acid, hydrogen bond acceptor, and hydrogen bond donor is 1:2:

6.

3. The ternary eutectic solvent containing bosonicine as described in claim 1, characterized in that: The hydrogen bond acceptor is betaine, and the hydrogen bond donor is xylitol.

4. The method for preparing a ternary eutectic solvent containing Bosein as described in claim 1, characterized in that, Including the following steps: Take appropriate molar ratios of Bosein, hydrogen bond donor, and hydrogen bond acceptor substances, and stir and melt them at a set temperature until they become a clear and transparent liquid, thus obtaining a ternary eutectic solvent containing Bosein.

5. The method for preparing a ternary eutectic solvent containing bosonicine as described in claim 4, characterized in that: The set temperature is 40–110°C.

6. The application of the ternary eutectic solvent containing bosine as described in any one of claims 1 to 3 in any aspect of the preparation of skin care products, cosmetics or topical medicines.

7. The application as described in claim 6, characterized in that: The skin care or cosmetic product contains at least ergothioneine.

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