Carboxymethyl muscarine amino acid salts, their preparation, their compositions and applications

Abstract

This invention discloses carboxymethyl sucrose amino acid salts, their preparation, compositions, and applications. The salts are prepared from carboxymethyl sucrose and arginine, or carboxymethyl sucrose and glycine, through a chemical reaction. The resulting salts undergo a series of processes including primary alcohol precipitation, redissolution, secondary alcohol precipitation, redissolution, tertiary alcohol precipitation, soaking, pulverization, filtration, and drying to obtain carboxymethyl sucrose arginine and carboxymethyl sucrose glycine salts. A composition is prepared by mixing these two salts. Both salts and the composition significantly inhibit the activity of matrix metalloproteinase 1 and promote the expression of type I collagen, thus delaying aging by maintaining the stability of collagen morphology and structure in the extracellular matrix. The preparation method of this invention is simple and the process conditions are mild. The product has the advantages of being weakly acidic, gentle on the skin, highly safe, and having good anti-aging effects. It can be widely used in skin care cosmetics and has promising application prospects.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to carboxymethyl sucralose amino acid salts (also known as carboxymethyl trehalose amino acid salts), their preparation, compositions, and applications. More specifically, it relates to a carboxymethyl sucralose amino acid salt (carboxymethyl sucralose arginine salt and carboxymethyl sucralose glycine salt) with anti-aging effects, its preparation, compositions, and applications. Background Technology

[0002] In today's rapidly developing society, "appearance anxiety" has become a hot topic, leading to a gradual increase in the number of younger people seeking anti-aging products. Aging, also known as degeneration, is a complex and irreversible natural phenomenon. It is the process by which an organism gradually loses and degenerates its constituent substances, tissue structure, and physiological functions over time or with increasing age, eventually leading to death. Phenolic compounds, flavonoids, vitamins, and carotenoids are widely used in anti-aging cosmetics. Currently, there are few reports on the use of sugar derivatives and amino acid salts as cosmetic ingredients in anti-aging evaluations. Therefore, further research, development, and enrichment of mild and safe sugar derivatives and amino acid salts as anti-aging ingredients are necessary.

[0003] Studies have shown that carboxymethyl sucrose (CMT) can dynamically regulate skin SELS values ​​(skin smoothness, skin roughness, degree of stratum corneum exfoliation, and wrinkle degree) within 30 days of application, significantly promote the proliferation and differentiation of HSF cells, and significantly promote COL I expression in HSF cells (p<0.05). Within a certain dosage range, it significantly inhibits MMP-1 expression, indicating that CMT can delay aging by maintaining the stability of collagen morphology and structure in the extracellular matrix (ECM). However, its application is limited due to the acidic nature of its aqueous solution, which is not gentle enough on the skin.

[0004] It is well known that Japan is at the forefront of international research on amino acids. In recent years, many Japanese patents have mentioned using arginine complex salts as active ingredients in various cosmetics. For example, some skin care products containing arginine complex salts have the functions of moisturizing, wrinkle removal, improving cracked skin, and making the skin elastic and shiny. Other studies have shown that glycine at a concentration of 1mM can increase collagen production.

[0005] Therefore, developing a carboxymethyl sucrose amino acid salt with anti-aging functions to expand the range of anti-aging raw materials is of great practical significance. Summary of the Invention

[0006] Due to the aforementioned deficiencies in existing technologies, this invention provides a carboxymethyl muscarinic acid salt with anti-aging functions to expand anti-aging raw materials. Specifically, it is a carboxymethyl muscarinic acid salt with anti-aging effects (a sugar derivative amino acid salt with strong anti-aging effects, specifically carboxymethyl muscarinic acid arginine salt and carboxymethyl muscarinic acid salt), its preparation, its composition, and its application. This invention uses carboxymethyl muscarinic acid as a base material, introduces arginine and glycine, and synthesizes a novel carboxymethyl muscarinic acid salt. The aqueous solution of this type of substance is gentler on the skin, more stable in finished cosmetic products, and is expected to achieve further enhancement on the basis of the original efficacy.

[0007] Existing research rarely mentions the formation of salts from carboxymethyl sucrose with other substances to alter the acidity or alkalinity of its aqueous solution. This invention reacts carboxymethyl sucrose with arginine and with glycine to obtain salts, and then combines the two salts in a certain proportion. The resulting carboxymethyl sucrose arginine salt, carboxymethyl sucrose glycine salt, and the combination of the two salts are used as the starting point for research and related tests on the inhibition of matrix metalloproteinase-1 (MMP-1) and the promotion of type I collagen expression, with the aim of delaying aging by maintaining the stability of collagen morphology and structure in the extracellular matrix (ECM).

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] Carboxymethyl muscarine amino acid salt, wherein the carboxymethyl muscarine amino acid salt is prepared from carboxymethyl muscarine and amino acids, and the carboxymethyl muscarine amino acid salt has the characteristics of inhibiting matrix metalloproteinase 1 (MMP-1) and promoting type I collagen expression, wherein the amino acid is arginine or glycine.

[0010] The preparation process of the above-mentioned carboxymethyl muscarine amino acid salt includes reaction, primary alcohol precipitation, resolution, secondary alcohol precipitation, resolution, tertiary alcohol precipitation, soaking, pulverization, filtration, and drying.

[0011] As a preferred technical solution:

[0012] The carboxymethyl sucrose amino acid salt described above, wherein the carboxymethyl sucrose amino acid salt is carboxymethyl sucrose arginine salt, has the following structural formula:

[0013] .

[0014] The carboxymethyl sucralose amino acid salt described above, wherein the methyl sucralose amino acid salt is a carboxymethyl sucralose glycine salt, has the following structural formula:

[0015] .

[0016] This invention provides a method for preparing the carboxymethyl muscarinic acid salt as described above, comprising the following steps:

[0017] (1) Weigh carboxymethyl muscarotene and amino acids in a molar ratio of 1:2, add deionized water to dissolve and react. The amino acid is arginine or glycine. The ratio of the sum of the mass of carboxymethyl muscarotene and amino acids to the mass of deionized water is 1:1 to 1:10. The reaction temperature is room temperature and the time is 1 to 6 hours.

[0018] (2) Slowly add 10 to 70 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at a speed of 200 to 800 rpm, let stand for 1 to 6 hours, discard the supernatant, and obtain the lower precipitate.

[0019] (3) Redissolve the precipitate obtained in step (2) with 1 to 10 times its mass of deionized water;

[0020] (4) Slowly add 10 to 70 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring while adding at a speed of 200 to 800 rpm, let stand for 1 to 6 hours, discard the supernatant, and obtain the lower precipitate;

[0021] (5) Redissolve the precipitate obtained in step (4) with 1 to 10 times its mass of deionized water;

[0022] (6) Slowly add 10 to 70 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at a speed of 200 to 800 rpm, and let stand for 12 to 24 hours to precipitate.

[0023] (7) The precipitate that has formed in the solution obtained in step (6) is crushed;

[0024] (8) Filter the solution obtained in step (7) using a microporous membrane and collect the precipitate. The pore size of the microporous membrane is 0.22 μm.

[0025] (9) The precipitate obtained in step (8) is dried under vacuum at 40~80℃ to obtain carboxymethyl muscarine amino acid salt.

[0026] The present invention also provides a carboxymethyl sucrose amino acid salt composition comprising the carboxymethyl sucrose arginine salt and the carboxymethyl sucrose glycine salt as described above, wherein the molar ratio of the carboxymethyl sucrose arginine salt to the carboxymethyl sucrose glycine salt is 1:6 to 6:1.

[0027] As a preferred technical solution:

[0028] The carboxymethyl sucrose amino acid salt composition described above, wherein the molar ratio of carboxymethyl sucrose arginine salt to carboxymethyl sucrose glycine salt is 1:3 to 3:1.

[0029] The carboxymethyl sucrose amino acid salt composition described above, wherein the molar ratio of carboxymethyl sucrose arginine salt to carboxymethyl sucrose glycine salt is 1:3, 3:1, or 1:1.

[0030] The present invention also provides the application of the carboxymethyl sucrose arginine salt, the carboxymethyl sucrose glycine salt, and the carboxymethyl sucrose amino acid salt composition as described above in inhibiting matrix metalloproteinase 1 (MMP-1) activity and promoting the expression of type I collagen.

[0031] The anti-aging activity of the combination of carboxymethyl sucrose arginine salt, carboxymethyl sucrose glycine salt, and carboxymethyl sucrose amino acid salt was characterized by the inhibition rate of matrix metalloproteinase 1 (MMP-1) and the relative expression level of type I collagen as test indicators.

[0032] Furthermore, the present invention also provides the use of the carboxymethyl sucrose arginine salt, the carboxymethyl sucrose glycine salt, and the carboxymethyl sucrose amino acid salt compositions described above in the preparation of cosmetics.

[0033] As a preferred technical solution:

[0034] As described above, the cosmetic is an aqueous solution, emulsion, spray, cream, gel, or mask.

[0035] The above technical solution is only one feasible technical solution of the present invention. The scope of protection of the present invention is not limited thereto. Those skilled in the art can reasonably adjust the specific design according to actual needs.

[0036] While some progress has been made in the study of the efficacy of carboxymethyl sucrose both domestically and internationally, its pH value tends to be acidic after dissolving in water. This invention modifies it by introducing amino acids to make it milder, more stable in cosmetic formulations, and more widely applicable dosage forms. In addition, previous research on amino acid complex salts has focused on the pharmaceutical field, where their applications are very extensive, and researchers have conducted many clinical applications and studies on their mechanisms of action. However, their application in cosmetics and health products is rare. This invention broadens the application of amino acid complex salts in other fields.

[0037] This invention tested the functionality of carboxymethyl sucrose arginine salt, carboxymethyl sucrose glycine salt, and carboxymethyl sucrose amino acid salt compositions from different perspectives (including target molecule cytotoxicity and anti-aging), filling the gap in efficacy research of sugar derivative amino acid salt compounds, thus giving sugar derivative amino acid salt compounds more application value; in addition, patch tests confirmed that they are safe and non-irritating compounds and compositions, meeting the current demand for active ingredients and effects in cosmetics.

[0038] The above invention has the following advantages or beneficial effects:

[0039] This invention is the first to synthesize carboxymethyl sucrose arginine salt and carboxymethyl sucrose glycine salt, and then combine them with carboxymethyl sucrose amino acid salt compositions. All three carboxymethyl sucrose arginine salt, carboxymethyl sucrose glycine salt, and carboxymethyl sucrose amino acid salt compositions exhibit superior anti-aging activity, showing good inhibitory effects on matrix metalloproteinase 1 (MMP-1) and promoting the expression of type I collagen, providing a material basis for the development of novel anti-aging cosmetics. The solvents and materials used in the preparation method of this invention are all industrial-grade, and the process is easy to control and repeat, reducing environmental impact and meeting the requirements of green chemistry and sustainable development. The preparation method of this invention has the potential for large-scale application, significant economic benefits, and good application prospects. Attached Figure Description

[0040] The invention, its features, shape, and advantages will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings. Like reference numerals denote like parts throughout the drawings. The drawings are not drawn to scale; their focus is on illustrating the gist of the invention.

[0041] Figure 1 The TGA chromatogram of carboxymethyl muscarin;

[0042] Figure 2 The DSC analysis chromatogram of carboxymethyl muscarin;

[0043] Figure 3 The TGA analysis chromatogram of arginine;

[0044] Figure 4 The DSC analysis spectrum of arginine;

[0045] Figure 5 The TGA chromatogram of glycine;

[0046] Figure 6 The DSC analysis spectrum of glycine;

[0047] Figure 7 TGA analysis chromatogram of carboxymethyl sucrose arginine salt;

[0048] Figure 8 DSC analysis chromatogram of carboxymethyl sucrose arginine salt;

[0049] Figure 9 TGA analysis chromatogram of carboxymethyl muscarinic acid glycinate;

[0050] Figure 10 The DSC chromatogram of carboxymethyl muscarinic acid glycine salt is shown.

[0051] Figure 11 This is a patch test image of a carboxymethyl muscarinic acid salt composition on human skin. Detailed Implementation

[0052] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but these are not intended to limit the scope of the invention.

[0053] The test methods for the compositions involved in the following examples are as follows:

[0054] Cytotoxicity assays include the following steps:

[0055] (1) Cell culture

[0056] HSF cells were cultured in DMEM / F12 (1:1) cell culture medium containing 15% fetal bovine serum and placed in a cell culture incubator at 37°C with 5% CO2. The medium was changed every 1–2 days. After digestion with 0.25% trypsin and centrifugation at 1200 rpm for 3–10 min, the supernatant was discarded, and fresh culture medium was added for passage culture.

[0057] (2) Preparation of sample solution of carboxymethyl muscarine amino acid salt composition

[0058] Dissolve and prepare 10 mg / mL carboxymethyl muscarine amino acid salt composition sample solutions in cell culture medium (the same as the culture medium in step 1), filter them through a bacterial membrane, and then dilute them to different concentrations according to the ratio.

[0059] (3) Cytotoxicity test

[0060] HSF cells were loaded at a rate of 1×10 5 Cells were seeded at a concentration of [cells / mL] in 96-well plates, with 100 μL of cell suspension added to each well. The 96-well plates were then incubated in an incubator for 24 hours (37°C, 5% CO2) to allow cell adhesion and growth. The supernatant was discarded, and 100 μL of different concentrations (5000, 2500, 1250, 625 μg / mL) of carboxymethyl muscarine amino acid salt combination solution was added to each well. The control group received 100 μL of blank cell culture medium. Six replicates were performed for each group. The 96-well plates were incubated in an incubator (37°C, 5% CO2) for another 24 hours. Then, 10 μL of CCK8 solution was added to each well. The 96-well plates were then incubated in an incubator (37°C, 5% CO2) for another 1 hour. The CCK8 kit used was purchased from Seven Innovations (Beijing) Biotechnology Co., Ltd.

[0061] (4) Data processing and analysis

[0062] The absorbance at 450 nm was measured using an ELISA reader. Cell viability was calculated using the following formula.

[0063]

[0064] The control group was tested without any sample added.

[0065] The determination of MMP-1 content in in vitro HSF cells includes the following steps:

[0066] (1) Cell inoculation

[0067] HSF cells were administered at a rate of 1.0 × 10⁻⁶. 5 The cells were seeded at a density of 100 μL / well and cultured at 37°C in a 5% CO2 cell culture incubator for about 24 hours.

[0068] (2) Preparation of carboxymethyl sucrose amino acid salt composition and carboxymethyl sucrose sample solution

[0069] The carboxymethyl sucrose amino acid salt composition and carboxymethyl sucrose were prepared by dissolving them in cell culture medium. After filtration through a bacterial membrane, they were diluted to 500 μg / mL sample solutions according to the specified ratio.

[0070] (3) Drug administration

[0071] Discard the supernatant, add 100 μL of the test sample with a concentration of 500 μg / mL to the well, and incubate at 37°C in a 5% CO2 cell culture incubator for 24 h;

[0072] (4) Collection of cell supernatant

[0073] After incubation, the cell culture supernatant was collected into a 1.5 mL sterile centrifuge tube and centrifuged at 1000×g, 4℃ for 20 min.

[0074] (5) MMP-1 inhibition rate determination

[0075] The detection was performed according to the instructions for use of the Human Matrix Metalloproteinase 1 Enzyme-Linked Immunosorbent Assay Kit, which was purchased from Jiangsu Edison Biotechnology Co., Ltd.

[0076] The determination of type I collagen content in HSF cells in vitro includes the following steps:

[0077] (1) Cell inoculation

[0078] HSF cells were administered at a rate of 1.0 × 10⁻⁶. 5 The cells were seeded at a density of 100 μL / well and cultured at 37°C in a 5% CO2 cell culture incubator for about 24 hours.

[0079] (2) Preparation of carboxymethyl sucrose amino acid salt composition and carboxymethyl sucrose sample solution

[0080] Carboxymethyl muscarinic acid salt composition and carboxymethyl muscarinic acid were prepared by dissolving them in cell culture medium, respectively. After filtration through a bacterial membrane, they were diluted to 500 μg / mL sample solutions according to the specified ratio.

[0081] (3) Drug administration

[0082] Discard the supernatant, add 100 μL of the test sample with a concentration of 500 μg / mL to the well, and incubate at 37°C in a 5% CO2 cell culture incubator for 24 h;

[0083] (4) Collection of cell supernatant

[0084] After incubation, the cell culture supernatant was collected into a 1.5 mL sterile centrifuge tube and centrifuged at 1000×g, 4℃ for 20 min.

[0085] (5) Determination of type I collagen content

[0086] The test was performed according to the instructions for use of the Human Type I Collagen Enzyme-Linked Immunosorbent Assay Kit, which was purchased from Jiangsu Edison Biotechnology Co., Ltd.

[0087] Safety patch testing includes the following steps:

[0088] Add 20 μL of the test solution to the spot tester, with the control well being a blank control (pure water). Apply the spot tester containing the test substance to the flexor side of the subject's forearm and gently press it with the palm of your hand to ensure even application to the skin for 24 hours. Observe the skin irritation and sensitization according to Table 1 at 30 min, 24 h, and 48 h after removing the spot tester, and record the results.

[0089] Table 1. Grading Criteria for Skin Reactions in Occlusive Patch Tests

[0090]

[0091] Example 1

[0092] The preparation method of carboxymethyl sucrose arginine salt includes the following steps:

[0093] (1) Weigh carboxymethyl muscarotene and arginine in a molar ratio of 1:2, add deionized water to dissolve and react. The ratio of the sum of the masses of carboxymethyl muscarotene and arginine to the mass of deionized water is 1:10. The reaction temperature is room temperature and the reaction time is 1h.

[0094] (2) Slowly add 70 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at 800 rpm, let stand for 6 hours, discard the supernatant, and obtain the lower precipitate.

[0095] (3) The precipitate obtained in step (2) is re-dissolved with 10 times its mass of deionized water;

[0096] (4) Slowly add 70 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring while adding at 800 rpm, let stand for 6 hours, discard the supernatant, and obtain the lower precipitate.

[0097] (5) Redissolve the precipitate obtained in step (4) with 10 times its mass of deionized water;

[0098] (6) Slowly add 70 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at 800 rpm, and let it stand for 24 hours to precipitate.

[0099] (7) Use a mixer to crush the precipitate that has formed in the solution obtained in step (6);

[0100] (8) Filter the solution obtained in step (7) using a 0.22 μm microporous membrane and collect the precipitate;

[0101] (9) The precipitate obtained in step (8) was vacuum dried at 40°C to obtain carboxymethyl sucrose arginine salt.

[0102] The elemental analysis of the obtained carboxymethyl sucrose arginine salt is shown in Table 2 below. The mass fractions of C, H, O, and N were 99.106%, indicating high purity. The TGA and DSC spectra of carboxymethyl sucrose arginine salt are shown below. Figure 7 and 8 As shown, the TGA and DSC spectra of carboxymethyl sucrose are as follows: Figure 1 and 2 As shown, the TGA and DSC analysis spectra of arginine are as follows: Figure 3 and 4 As shown in the figure. Combining TGA and DSC graphs, it can be analyzed that the decomposition temperature of carboxymethyl sucrose is about 337℃, the decomposition temperature of arginine is about 244℃, and the decomposition temperature of carboxymethyl sucrose arginine salt is about 280℃. This shows a significant change compared to the decomposition temperatures of carboxymethyl sucrose and arginine, indicating salt formation.

[0103] Table 2. Elemental analysis of methyl sucrose arginine salts

[0104]

[0105] Example 2

[0106] The preparation method of carboxymethyl sucrose arginine salt includes the following steps:

[0107] (1) Weigh carboxymethyl muscarotene and arginine in a molar ratio of 1:2, add deionized water to dissolve and react. The ratio of the sum of the masses of carboxymethyl muscarotene and arginine to the mass of deionized water is 1:1. The reaction temperature is room temperature and the reaction time is 6h.

[0108] (2) Slowly add 10 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at 200 rpm, let stand for 1 hour, discard the supernatant, and obtain the lower precipitate.

[0109] (3) Redissolve the precipitate obtained in step (2) with 1 times its mass of deionized water;

[0110] (4) Slowly add 10 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring at 200 rpm, let stand for 1 hour, discard the supernatant, and obtain the lower precipitate.

[0111] (5) Redissolve the precipitate obtained in step (4) with 1 times its mass of deionized water;

[0112] (6) Slowly add 10 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at a speed of 200 rpm, and let it stand for 12 hours to precipitate.

[0113] (7) Use a mixer to crush the precipitate that has formed in the solution obtained in step (6);

[0114] (8) Filter the solution obtained in step (7) using a 0.22 μm microporous membrane and collect the precipitate;

[0115] (9) The precipitate obtained in step (8) is dried under vacuum at 80°C to obtain carboxymethyl sucrose arginine salt.

[0116] Example 3

[0117] The preparation method of carboxymethyl muscarinic acid salt includes the following steps:

[0118] (1) Weigh carboxymethyl muscarotene and glycine in a molar ratio of 1:2, add deionized water to dissolve and react. The ratio of the sum of the masses of carboxymethyl muscarotene and glycine to the mass of deionized water is 1:10. The reaction temperature is room temperature and the reaction time is 1 h.

[0119] (2) Slowly add 70 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at 800 rpm, let stand for 6 hours, discard the supernatant, and obtain the lower precipitate.

[0120] (3) The precipitate obtained in step (2) is re-dissolved with 10 times its mass of deionized water;

[0121] (4) Slowly add 70 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring while adding at 800 rpm, let stand for 6 hours, discard the supernatant, and obtain the lower precipitate.

[0122] (5) Redissolve the precipitate obtained in step (4) with 10 times its mass of deionized water;

[0123] (6) Slowly add 70 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at 800 rpm, and let it stand for 24 hours to precipitate.

[0124] (7) Use a mixer to crush the precipitate that has formed in the solution obtained in step (6);

[0125] (8) Filter the solution obtained in step (7) using a 0.22 μm microporous membrane and collect the precipitate;

[0126] (9) The precipitate obtained in step (8) was dried under vacuum at 40°C to obtain carboxymethyl muscarinic acid salt.

[0127] The elemental analysis of the prepared carboxymethyl sucrose glycinate is shown in Table 3. The mass fractions of C, H, O, and N were 99.071%, indicating high purity. The TGA and DSC spectra of the carboxymethyl sucrose glycinate are shown in Table 3. Figure 9 and 10 As shown, the TGA and DSC spectra of carboxymethyl sucrose are as follows: Figure 1 and 2 As shown, the TGA and DSC analysis spectra of glycine are as follows: Figure 5 and 6 As shown in the figure. Combining TGA and DSC graphs, it can be analyzed that the decomposition temperature of carboxymethyl muscarotene is about 337℃, the decomposition temperature of glycine is about 250~260℃, while the decomposition temperature of carboxymethyl muscarotene glycinate is about 215~230℃. This shows a significant change compared to the decomposition temperatures of carboxymethyl muscarotene and glycine, indicating salt formation.

[0128] Table 3. Elemental analysis of methyl muscarinic acid salts

[0129]

[0130] Example 4

[0131] The preparation method of carboxymethyl muscarinic acid salt includes the following steps:

[0132] (1) Weigh carboxymethyl muscarotene and glycine in a molar ratio of 1:2, add deionized water to dissolve and react. The ratio of the sum of the masses of carboxymethyl muscarotene and glycine to the mass of deionized water is 1:1. The reaction temperature is room temperature and the reaction time is 6h.

[0133] (2) Slowly add 10 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at 200 rpm, let stand for 1 hour, discard the supernatant, and obtain the lower precipitate.

[0134] (3) Redissolve the precipitate obtained in step (2) with 1 times its mass of deionized water;

[0135] (4) Slowly add 10 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring at 200 rpm, let stand for 1 hour, discard the supernatant, and obtain the lower precipitate.

[0136] (5) Redissolve the precipitate obtained in step (4) with 1 times its mass of deionized water;

[0137] (6) Slowly add 10 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at a speed of 200 rpm, and let it stand for 12 hours to precipitate.

[0138] (7) Use a mixer to crush the precipitate that has formed in the solution obtained in step (6);

[0139] (8) Filter the solution obtained in step (7) using a 0.22 μm microporous membrane and collect the precipitate;

[0140] (9) The precipitate obtained in step (8) is dried under vacuum at 80°C to obtain carboxymethyl muscarinic acid salt.

[0141] Example 5

[0142] The preparation method of the carboxymethyl muscarinic acid salt composition is as follows: the carboxymethyl muscarinic arginine salt obtained in Example 1 and the carboxymethyl muscarinic glycine salt obtained in Example 3 are mixed and pulverized according to a certain molar ratio, and then mixed evenly to obtain the carboxymethyl muscarinic acid salt composition.

[0143] Compositions 1-15 and comparative examples 1-3 were prepared using the above method according to the proportions in Table 4.

[0144] Table 4 Composition of each composition and comparative formulation

[0145]

[0146] The cytotoxicity test results of compositions 1-15 and comparative examples 1-3 are shown in Table 5.

[0147] Table 5. Cytotoxicity test results of compositions 1-15 and comparative examples 1-3

[0148]

[0149]

[0150]

[0151] The MMP-1 inhibition rates of the blank control group, compositions 1-15, and comparative examples 1-3 are shown in Table 6.

[0152] Table 6. MMP-1 inhibition rates of compositions 1-15 and comparative examples 1-3

[0153]

[0154] The relative expression levels of COL I in the blank control group, compositions 1-15, and comparative examples 1-3 are shown in Table 7.

[0155] Table 7. Relative expression levels of COL I in compositions 1-15 and comparative examples 1-3

[0156]

[0157] By observing the performance comparison of the composition and the comparative example, it can be seen that the carboxymethyl muscarinic acid salt composition is significantly better than single carboxymethyl muscarinic acid salts and carboxymethyl muscarin.

[0158] Human skin patch tests were performed on compositions 10, 14, and 15. The results are shown in Table 8. Figure 11 Human skin patch test results showed no adverse skin reactions in 30 participants.

[0159] Table 8 Results of Cosmetic Human Skin Patch Test

[0160]

[0161] Those skilled in the art should understand that variations can be implemented by combining existing technology with the above embodiments, which will not be elaborated here. Such variations do not affect the essence of the present invention, and will not be elaborated here either.

[0162] The preferred embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and the devices and structures not described in detail should be understood as being implemented in a conventional manner in the art. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention using the methods and techniques disclosed above, or modify them into equivalent embodiments with equivalent changes, without departing from the scope of the present invention. This does not affect the essential content of the present invention. Therefore, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the present invention's technical solutions still fall within the protection scope of the present invention.

Claims

1. A carboxymethyl sucrose amino acid salt, characterized in that, The carboxymethyl sucrose amino acid salt is prepared from carboxymethyl sucrose and amino acids, and the carboxymethyl sucrose amino acid salt has the characteristics of inhibiting matrix metalloproteinase 1 and promoting the expression of type I collagen. The amino acid is arginine or glycine.

2. The carboxymethyl sucrose amino acid salt according to claim 1, characterized in that, The carboxymethyl sucrose amino acid salt is carboxymethyl sucrose arginine salt, and its structural formula is as follows: 。 3. The carboxymethyl sucrose amino acid salt according to claim 1, characterized in that, The carboxymethyl muscarinic acid salt is carboxymethyl muscarinic glycine salt, and its structural formula is as follows: 。 4. The method for preparing carboxymethyl sucrose amino acid salt according to any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Weigh carboxymethyl muscarotene and amino acids in a molar ratio of 1:2, add deionized water to dissolve and react. The amino acid is arginine or glycine. The ratio of the sum of the mass of carboxymethyl muscarotene and amino acids to the mass of deionized water is 1:1 to 1:

10. The reaction temperature is room temperature and the time is 1 to 6 hours. (2) Slowly add 10 to 70 times the volume of anhydrous ethanol to the reaction solution obtained in step (1), stirring while adding at a speed of 200 to 800 rpm, let stand for 1 to 6 hours, discard the supernatant, and obtain the lower precipitate. (3) Redissolve the precipitate obtained in step (2) with 1 to 10 times its mass of deionized water; (4) Slowly add 10 to 70 times the volume of anhydrous ethanol to the solution obtained in step (3), stirring while adding at a speed of 200 to 800 rpm, let stand for 1 to 6 hours, discard the supernatant, and obtain the lower precipitate; (5) Redissolve the precipitate obtained in step (4) with 1 to 10 times its mass of deionized water; (6) Slowly add 10 to 70 times the volume of anhydrous ethanol to the solution obtained in step (5), stirring while adding at a speed of 200 to 800 rpm, and let stand for 12 to 24 hours to precipitate. (7) The precipitate that has formed in the solution obtained in step (6) is crushed; (8) Filter the solution obtained in step (7) using a microporous membrane and collect the precipitate. The pore size of the microporous membrane is 0.22 μm. (9) The precipitate obtained in step (8) is dried under vacuum at 40~80℃ to obtain carboxymethyl muscarine amino acid salt.

5. A carboxymethyl sucrose amino acid salt composition, characterized in that, It includes the carboxymethyl sucrose arginine salt as described in claim 2 and the carboxymethyl sucrose glycine salt as described in claim 3, wherein the molar ratio of the carboxymethyl sucrose arginine salt to the carboxymethyl sucrose glycine salt is 1:6 to 6:

1.

6. The carboxymethyl sucrose amino acid salt composition according to claim 5, characterized in that, The molar ratio of carboxymethyl sucrose arginine salt to carboxymethyl sucrose glycine salt is 1:3 to 3:

1.

7. The carboxymethyl sucrose amino acid salt composition according to claim 5, characterized in that, The molar ratio of carboxymethyl sucrose arginine salt to carboxymethyl sucrose glycine salt is 1:3, 3:1, or 1:

1.

8. The use of the carboxymethyl sucrose arginine salt as described in claim 2, the carboxymethyl sucrose glycine salt as described in claim 3, and the carboxymethyl sucrose amino acid salt composition as described in any one of claims 5 to 7 in inhibiting matrix metalloproteinase 1 activity and promoting the expression of type I collagen.

9. The use of the carboxymethyl sucrose arginine salt as described in claim 2, the carboxymethyl sucrose glycine salt as described in claim 3, and the carboxymethyl sucrose amino acid salt composition as described in any one of claims 5 to 7 in the preparation of cosmetics.

10. The application according to claim 9, characterized in that, The cosmetics are aqueous solutions, emulsions, sprays, creams, gels, or masks.

Images