Silk fibroin aggregate-based dispersed organic active substance as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a silk fibroin aggregate-based dispersed organic active substance as well as a preparation method and application thereof. The preparation method of the silk fibroin aggregate-based dispersed organic active substance comprises the following steps: (1) dissolving an organic active substance in a first solvent to obtain a first solution; dissolving the silk fibroin aggregate in a second solvent to obtain a second solution; (2) slowly adding the second solution into the first solution, mixing and homogenizing to obtain a silk fibroin aggregate-based dispersed organic active substance; wherein the silk fibroin aggregate is prepared by mixing a silk fibroin aqueous solution and alcohol. The silk fibroin aggregate-based dispersed organic active substance prepared by the preparation method can improve the bioavailability of the organic active substance (such as vitamin C, hyaluronic acid, coenzyme Q10, nicotinamide, arbutin, kojic acid, licorice extract, centella extract, chamomile extract or green tea extract) in cosmetics.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to a silk fibroin aggregate-based dispersed organic active substance, a preparation method and an application thereof. Background Art

[0002] Organic active substances (such as vitamin C, polyphenols, carotenoids, etc.) are core ingredients in cosmetics and have important application value in scenarios such as anti-oxidation, anti-photoaging, and whitening. However, these substances have significant defects in actual formulations: due to the stratum corneum barrier and the hydrophilic and hydrophobic properties of the active ingredients themselves, less than 15% of the active ingredients in conventional formulas can be absorbed and utilized by the skin. This dual bottleneck of stability and transdermal efficiency seriously restricts the full realization of the efficacy of cosmetics.

[0003] The industry currently employs technologies such as microencapsulation, liposome encapsulation, or polymer nanocarriers to address these issues. For example, while liposome encapsulation can enhance the stability of some active ingredients, its preparation relies on additives such as surfactants and cholesterol, which can cause skin irritation. Furthermore, nanocarrier technologies based on synthetic polymers often suffer from a mismatch between the degradation rate of the carrier material and the release kinetics of the active ingredient, leading to a burst release effect or ineffective sustained release.

[0004] In addition, existing technologies generally have industrial barriers such as complex processes and high costs, and most carrier systems find it difficult to simultaneously achieve long-term protection and efficient delivery of active ingredients. In particular, when dealing with the need to co-load multiple active ingredients with significant differences in polarity, the compatibility is significantly insufficient. Summary of the Invention

[0005] The present invention provides a preparation method and application of a silk fibroin aggregate-based dispersed organic active substance. By combining the organic active substance with the silk fibroin aggregate, a complex with good dispersibility and stability is prepared, which significantly improves the bioavailability and stability of the organic active substance in cosmetics and has broad application prospects.

[0006] The present invention aims to provide a method for preparing and applying a silk fibroin aggregate-based dispersion of an organic active substance. By combining the organic active substance with silk fibroin aggregates, a composite with excellent dispersibility and stability is prepared, thereby improving the bioavailability and stability of the organic active substance in cosmetics. Currently, no effective solution has been proposed to address the problems of poor dispersion uniformity, complex preparation steps, poor stability, and low bioavailability in related technologies.

[0007] In order to achieve the above object, the present invention can adopt the following technical solutions:

[0008] In one aspect, the present invention provides a method for preparing a silk fibroin aggregate-based dispersed organic active substance, the preparation method comprising:

[0009] (1) dissolving an organic active substance in a first solvent to obtain a first solution; and dissolving a silk fibroin aggregate in a second solvent to obtain a second solution;

[0010] (2) slowly adding the second solution into the first solution and mixing them to homogenize the silk fibroin aggregate-based dispersed organic active substance;

[0011] The silk fibroin aggregate is prepared by mixing a silk fibroin aqueous solution with alcohol.

[0012] Preferably, in the above preparation method,

[0013] The organic active substance is selected from one or more combinations of vitamin C, vitamin E, hyaluronic acid, coenzyme Q10, ceramide, niacinamide, arbutin, kojic acid, licorice extract, centella asiatica extract, chamomile extract, green tea extract, grape seed extract, squalane, tocopherol, retinol, retinol derivatives, peptide active substances, phytosterols or tetrahydromagnanol; and / or

[0014] The first solvent and the second solvent are independently selected from one or more combinations of water, ethanol, propylene glycol or glycerol.

[0015] Preferably, the above preparation method satisfies one or more of the following conditions:

[0016] (i) In step (2), stirring is performed during mixing at a speed of 100 rpm to 1000 rpm for a time of 10 min to 60 min;

[0017] (ii) In step (2), the homogenization pressure is 100 bar to 500 bar, and the homogenization time is 5 min to 30 min.

[0018] Another aspect of the present invention provides a silk fibroin aggregate-based dispersed organic active substance prepared by the preparation method of the present invention.

[0019] In another aspect, the present invention provides a cosmetic comprising the silk fibroin aggregate-based dispersed organic active substance of the present invention.

[0020] Preferably, the cosmetics are lotion, essence, cream, mask, spray, foundation, lipstick, eye shadow or blush.

[0021] Preferably, the mass fraction of the silk fibroin aggregate-based dispersed organic active substance in the cosmetic is 0.1 wt%-10 wt%.

[0022] The beneficial effects of the present invention include: the silk fibroin aggregate-based dispersed organic active substances provided by the present invention can improve the bioavailability of organic active substances (such as vitamin C, hyaluronic acid, coenzyme Q10, niacinamide, arbutin, kojic acid, licorice extract, Centella asiatica extract, chamomile extract or green tea extract) in cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a digital photograph of the silk fibroin fiber aggregate dispersion emulsion of the present invention;

[0024] Figure 2 This is a scanning electron micrograph of the silk fibroin fiber aggregate dispersion emulsion of the present invention (scale bar: 100 μm);

[0025] Figure 3 is a dynamic light scattering (DLS) graph of the silk fibroin fiber aggregate dispersion emulsion of the present invention;

[0026] Figure 4 It is the Zeta potential diagram of the silk fibroin fiber aggregate dispersion emulsion of the present invention. DETAILED DESCRIPTION

[0027] The examples are provided to better illustrate the present invention, but are not intended to limit the present invention to the examples. Therefore, non-essential improvements and adjustments to the embodiments made by those skilled in the art based on the above-mentioned invention still fall within the scope of protection of the present invention.

[0028] The terms used herein are only used to describe specific embodiments and are not intended to limit the present disclosure. Unless the context has a significantly different meaning, expressions in the singular include expressions in the plural. As used herein, it should be understood that terms such as "include", "have", "comprise" and the like are intended to indicate the presence of features, numbers, operations, components, parts, elements, materials or combinations. The terms of the present invention are disclosed in the specification and are not intended to exclude the possibility that one or more other features, numbers, operations, components, parts, elements, materials or combinations thereof may exist or may be added. As used herein, " / " may be interpreted as "and" or "or", depending on the circumstances.

[0029] The present embodiment provides a method for preparing a silk fibroin aggregate-based dispersed organic active substance, the preparation method comprising: (1) dissolving the organic active substance in a first solvent to obtain a first solution; dissolving the silk fibroin aggregate in a second solvent to obtain a second solution; (2) slowly adding the second solution to the first solution, mixing, and homogenizing to obtain the silk fibroin aggregate-based dispersed organic active substance; wherein the silk fibroin aggregate is prepared by mixing a silk fibroin aqueous solution with alcohol.

[0030] In some specific examples, in the above preparation method, the organic active substance is selected from one or more combinations of vitamin C, vitamin E, hyaluronic acid, coenzyme Q10, ceramide, niacinamide, arbutin, kojic acid, licorice extract, Centella asiatica extract, chamomile extract, green tea extract, grape seed extract, squalane, tocopherol, retinol, retinol derivatives, peptide active substances, phytosterols or tetrahydromagnanol; and / or the first solvent and the second solvent are independently selected from one or more combinations of water, ethanol, propylene glycol or glycerol.

[0031] It should be noted that, in the above preparation method, the organic active substance is well known in the art; in addition, the first solvent is all solvents that can dissolve the organic active substance, such as one or more combinations of water, ethanol, propylene glycol or glycerol, and the second solvent is all solvents that can dissolve silk fibroin aggregates, such as one or more combinations of water, ethanol, propylene glycol or glycerol.

[0032] In some specific examples, the above preparation method satisfies one or more of the following conditions:

[0033] (i) In step (2), stirring is performed during mixing at a speed of 100 rpm to 1000 rpm (e.g., 300 rpm, 500 rpm, or 300 rpm) for 10 min to 60 min (e.g., 20 min, 30 min, 40 min, or 50 min);

[0034] (ii) In step (2), the homogenization pressure is 100 bar to 500 bar (e.g., 200 bar, 300 bar, or 400 bar, etc.), and the homogenization time is 5 min to 30 min (e.g., 10 min, 15 min, 20 min, or 25 min, etc.).

[0035] The embodiment of the present invention further provides a silk fibroin aggregate-based dispersed organic active substance prepared by the preparation method of the present invention.

[0036] It should be noted that the silk fibroin aggregate-based dispersed organic active substances prepared in the present invention can improve the bioavailability of the organic active substances in cosmetics.

[0037] An embodiment of the present invention further provides a cosmetic comprising the silk fibroin aggregate-based dispersed organic active substance of the present invention.

[0038] In some specific examples, the cosmetics are lotion, essence, cream, mask, spray, foundation, lipstick, eye shadow or blush.

[0039] In some specific examples, the mass fraction of the silk fibroin aggregate-based dispersed organic active substance in the cosmetic is 0.1 wt%-10 wt%, for example, 1 wt%, 3 wt%, 5 wt%, 7 wt% or 9 wt%.

[0040] From the above, it can be seen that compared with gelatin and collagen, silk fibroin has a unique molecular structure and excellent performance. Its molecular structure contains a large number of repetitive sequences and rich amino acid composition, which can form a close interaction with organic active substances, effectively improving the dispersibility and stability of organic active substances. Silk fibroin has good solubility and dispersibility. Through the preparation method of the present invention, it can be evenly mixed with organic active substances to form a stable complex. For example, in Example 1, the bioavailability of vitamin C in the emulsion after silk fibroin treatment was increased by 30%, which is significantly higher than the effect of gelatin and collagen treatment. In addition, silk fibroin also has good biocompatibility and moisturizing properties, which can further enhance the use effect and user experience of cosmetics. In terms of stability, silk fibroin aggregate-based dispersed organic active substances show good stability in different cosmetic formulas, are not easily affected by environmental factors, and effectively solve the stability problem of organic active substances in cosmetics.

[0041] In summary, silk fibroin has significant advantages in improving the bioavailability and stability of organic active substances, is superior to other types of protein processing methods, and can better meet the cosmetics industry's demand for the application of organic active substances.

[0042] In order to better understand the present invention, the content of the present invention is further explained below with reference to specific examples, but the content of the present invention is not limited to the following examples.

[0043] 1. Preparation of Silk Fibroin Aggregate-Based Dispersed Organic Active Substances

[0044] In the following example, the preparation method of silk fibroin aggregate (hereinafter also referred to as silk fibroin fiber aggregate dispersed emulsion) includes: dissolving 10g of silk fibroin (purchased from Suzhou Pinglan Biotechnology Co., Ltd.) in water, adding 500g of methanol and mixing evenly, standing for 30min, centrifuging at 3000rpm for 5min to obtain silk fibroin aggregate.

[0045] Preparation Example 1

[0046] (1) dissolving 0.5 g of vitamin C in 10 mL of ethanol to obtain a first solution;

[0047] (2) dissolving 1 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0048] (3) At 60°C, the second solution was slowly added to the first solution at a rate of 100 mL / min, and the mixture was stirred at a speed of 500 rpm for 30 min to obtain a mixed solution;

[0049] (4) The mixed solution was homogenized at a pressure of 300 bar for 15 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0050] Preparation Example 2

[0051] (1) dissolving 0.3 g of hyaluronic acid in 10 mL of glycerol to obtain a first solution;

[0052] (2) dissolving 1.5 g of silk fibroin aggregate in 10 mL of propylene glycol to obtain a second solution;

[0053] (3) At 70°C, the second solution was slowly added to the first solution at a rate of 50 mL / min, and the mixture was stirred at 800 rpm for 20 min to obtain a mixed solution;

[0054] (4) The mixed solution was homogenized at a pressure of 400 bar for 20 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0055] Preparation Example 3

[0056] (1) Dissolving 0.4 g of coenzyme Q10 in 10 mL of ethanol to obtain a first solution;

[0057] (2) dissolving 2 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0058] (3) At 50°C, the second solution was slowly added to the first solution at a rate of 80 mL / min, and the mixture was stirred at a speed of 600 rpm for 25 min to obtain a mixed solution;

[0059] (4) The mixed solution was homogenized at a pressure of 350 bar for 18 minutes to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0060] Preparation Example 4

[0061] (1) dissolving 0.6 g of nicotinamide in 10 mL of propylene glycol to obtain a first solution;

[0062] (2) dissolving 1.2 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0063] (3) At 80°C, the second solution was slowly added to the first solution at a rate of 120 mL / min, and the mixture was stirred at a speed of 700 rpm for 22 min to obtain a mixed solution;

[0064] The mixed solution was homogenized at a pressure of 450 bar for 25 minutes to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0065] Preparation Example 5

[0066] (1) dissolving 0.7 g of arbutin in 10 mL of glycerol to obtain a first solution;

[0067] (2) dissolving 1.8 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0068] (3) At 45°C, the second solution was slowly added to the first solution at a rate of 90 mL / min, and the mixture was stirred at a speed of 550 rpm for 28 min to obtain a mixed solution;

[0069] (4) The mixed solution was homogenized at a pressure of 320 bar for 12 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0070] Preparation Example 6

[0071] (1) dissolving 0.8 g of kojic acid in 10 mL of ethanol to obtain a first solution;

[0072] (2) dissolving 2.2 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0073] (3) At 75°C, the second solution was slowly added to the first solution at a rate of 110 mL / min, and the mixture was stirred at a speed of 650 rpm for 24 min to obtain a mixed solution;

[0074] (4) The mixed solution was homogenized at a pressure of 420 bar for 28 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0075] Preparation Example 7

[0076] (1) dissolving 0.9 g of licorice extract in 10 mL of propylene glycol to obtain a first solution;

[0077] (2) dissolving 2.5 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0078] (3) At 65°C, the second solution was slowly added to the first solution at a rate of 130 mL / min, and the mixture was stirred at a speed of 750 rpm for 26 min to obtain a mixed solution;

[0079] (4) The mixed solution was homogenized at a pressure of 480 bar for 30 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0080] Preparation Example 8

[0081] (1) dissolving 1.0 g of Centella asiatica extract in 10 mL of glycerol to obtain a first solution;

[0082] (2) dissolving 3.0 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0083] (3) At 55°C, the second solution was slowly added to the first solution at a rate of 140 mL / min, and the mixture was stirred at a speed of 850 rpm for 20 min to obtain a mixed solution;

[0084] (4) The mixed solution was homogenized at a pressure of 380 bar for 15 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0085] Preparation Example 9

[0086] (1) dissolving 1.1 g of chamomile extract in 10 mL of ethanol to obtain a first solution;

[0087] (2) dissolving 3.5 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0088] (3) At 85°C, the second solution was slowly added to the first solution at a rate of 150 mL / min, and the mixture was stirred at 900 rpm for 18 min to obtain a mixed solution;

[0089] (4) The mixed solution was homogenized at a pressure of 410 bar and a time of 22 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0090] Preparation Example 10

[0091] (1) dissolving 1.2 g of green tea extract in 10 mL of propylene glycol to obtain a first solution;

[0092] (2) dissolving 4.0 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0093] (3) At 40°C, the second solution was slowly added to the first solution at a rate of 160 mL / min, and the mixture was stirred at a speed of 500 rpm for 30 min to obtain a mixed solution;

[0094] (4) The mixed solution was homogenized at a pressure of 360 bar for 20 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0095] Preparation Example 11

[0096] (1) dissolving 1.2 g of retinol in 10 mL of propylene glycol to obtain a first solution;

[0097] (2) dissolving 4.0 g of silk fibroin aggregate in 10 mL of water to obtain a second solution;

[0098] (3) At 40°C, the second solution was slowly added to the first solution at a rate of 160 mL / min, and the mixture was stirred at a speed of 500 rpm for 30 min to obtain a mixed solution;

[0099] (4) The mixed solution was homogenized at a pressure of 360 bar for 20 min to obtain a silk fibroin aggregate-based dispersed organic active substance.

[0100] Preparation Comparative Example 1

[0101] (1) dissolving 0.5 g of vitamin C in 10 mL of ethanol to obtain a first solution;

[0102] (2) Dissolve 1 g of gelatin in 10 mL of water to obtain a second solution;

[0103] (3) At 60°C, the second solution was slowly added to the first solution at a rate of 100 mL / min, and the mixture was stirred at a speed of 500 rpm for 30 min to obtain a mixed solution;

[0104] (4) The mixed solution was homogenized at a pressure of 300 bar for 15 min to obtain a gelatin-based dispersed organic active substance.

[0105] Preparation Comparative Example 2

[0106] (1) dissolving 0.3 g of hyaluronic acid in 10 mL of glycerol to obtain a first solution;

[0107] (2) dissolving 1.5 g of collagen in 10 mL of water to obtain a second solution;

[0108] (3) At 70°C, the second solution was slowly added to the first solution at a rate of 50 mL / min, and the mixture was stirred at 800 rpm for 20 min to obtain a mixed solution;

[0109] (4) The mixed solution was homogenized at a pressure of 400 bar for 20 min to obtain a collagen-based dispersed organic active substance.

[0110] 2. Characterization of Silk Fibroin Aggregates

[0111] Digital photos of the silk fibroin aggregates of the present invention are as follows: Figure 1 shown.

[0112] The scanning electron micrograph (scale bar: 100 μm) of the silk fibroin aggregate of the present invention is as follows Figure 2 shown.

[0113] The dynamic light scattering (DLS) diagram of the silk fibroin aggregates of the present invention is as follows: Figure 3 shown.

[0114] The Zeta potential diagram of the silk fibroin aggregate and the silk fibroin aggregate-based dispersed organic active substance of the present invention is as follows: Figure 4 As shown, SF is a silk fibroin aqueous solution, SFN is a silk fibroin aggregate, Sample 1 refers to the silk fibroin aggregate-based dispersed organic active substance (hyaluronic acid) prepared in Example 1, and Sample 2 is the silk fibroin aggregate-based dispersed organic active substance (retinol) prepared in Example 11.

[0115] 3. Application of Silk Fibroin Aggregate-Based Dispersed Organic Active Substances

[0116] In the following example, the stability test method is as follows:

[0117] (1) Physical stability test

[0118] Centrifugal stability test: Place cosmetic samples such as lotions or essences in a centrifuge and centrifuge at a certain speed (3000 rpm) for a certain period of time (30 minutes) to observe whether the sample shows any signs of stratification or precipitation. If the sample remains homogeneous after centrifugation, it indicates good physical stability.

[0119] Long-term storage stability test: The samples were placed under different temperature (4°C, 25°C, 40°C) and humidity (20%, 40%, 60%) conditions, and their appearance, texture, color, etc. were regularly observed. The observation period was 6 months. If the samples did not show abnormal phenomena such as deterioration, delamination, color change, etc. during storage, they were considered to have good long-term stability.

[0120] Thermal stability test: Place the sample in a high temperature environment (45℃-60℃) for a certain period of time (2 weeks), then cool it to room temperature and observe whether it becomes thinner, thicker, or changes color. If the sample can still maintain its original state after high temperature treatment, it means that its thermal stability is good.

[0121] (2) Chemical stability test

[0122] pH stability test: Under different storage conditions, the pH value of the sample is regularly measured. If the pH value of the sample remains within a certain range during storage (the change does not exceed ±0.5), it indicates that its chemical stability is good;

[0123] Ingredient stability test: The changes in the content of key ingredients (such as vitamin C, hyaluronic acid, etc.) in the sample are determined by high-performance liquid chromatography (HPLC) analysis. If the decrease in the content of key ingredients during storage is within an acceptable range (such as no more than 10%), the sample is considered to have good chemical stability.

[0124] In the following example, the moisturizing effect is tested as follows:

[0125] (1) Skin moisture content test

[0126] Use a skin moisture tester (such as Corneometer CM825) to measure the skin moisture content of volunteers before and after using lotion or essence. The inner side of the volunteer's forearm is usually selected as the test site, and tests are conducted before use of the product and at time points such as 0.5 hours, 1 hour, 2 hours, and 4 hours after use. If the skin moisture content increases significantly after use of the product and remains at a high level for a certain period of time, it indicates that the product has a good moisturizing effect.

[0127] (2) Skin stratum corneum hydration test

[0128] A skin stratum corneum hydration tester (such as Tewameter TM300) is used to measure the hydration level of the skin's stratum corneum. The transepidermal water loss (TEWL) value on the skin surface is measured to indirectly reflect the skin's moisturizing status. The lower the TEWL value, the stronger the skin's moisturizing ability. During the test, the inner side of the volunteers' forearms are also selected as the test site. Tests are performed before and a certain period of time (2 hours and 4 hours) after using the product to compare changes in TEWL values.

[0129] In the following example, the bioavailability testing method includes: using an in vitro skin model (ex vivo pig skin model), applying an essence with added silk fibroin aggregate-based dispersed organic active substances and an essence without added substances to the surface of the skin model respectively, and after a certain period of time (24 hours), measuring the hyaluronic acid content in the skin model by high performance liquid chromatography (HPLC), comparing the difference in absorption amount between the two, and thus determining the effect of the silk fibroin aggregate-based dispersed organic active substances on the bioavailability of hyaluronic acid.

[0130] In the following example, the specific calculation method of bioavailability is as follows:

[0131] In vitro testing: Using a skin model (ex vivo pig skin model), the cosmetic sample is applied to the surface of the skin model. After a certain period of time, the content of the active ingredient in the skin model is determined by high-performance liquid chromatography (HPLC). The bioavailability (%) can be calculated using the following formula:

[0132] Bioavailability = (content of active ingredient in skin model / total amount of active ingredient in applied cosmetics) × 100%;

[0133] Calculation of improved bioavailability: The improved bioavailability can be calculated by comparing the bioavailability of the active ingredient in the cosmetics containing the silk fibroin aggregate-based dispersed organic active substance with that in the control cosmetics without the addition of the active ingredient. The specific calculation formula is as follows:

[0134] Percentage increase in bioavailability = (bioavailability of active ingredient in cosmetics containing silk fibroin aggregate-dispersed organic active substances - bioavailability of active ingredient in control cosmetics without such substances) / total amount of active ingredient in applied cosmetics × 100%; for example, in Application Example 1, the total amount of vitamin C in the applied emulsion is 100 mg, and after 24 hours, the vitamin C content detected in the skin model is 20 mg. The bioavailability of the vitamin C not treated with silk fibroin is: bioavailability = 20 / 100 × 100% = 20%; and the vitamin C content dispersed by silk fibroin aggregates detected in the skin model is 50 mg. The bioavailability of the vitamin C dispersed by silk fibroin aggregates is: bioavailability = 50 / 100 × 100% = 50%.

[0135] In the following example, the sensory evaluation includes: organizing 10 volunteers to use cosmetics containing silk fibroin aggregate-based dispersed organic active substances and a control product without such substances, and having the users evaluate their experience of using the products (such as moisturizing, absorption speed, skin condition after use, etc.); and further verifying the actual effect of the product by statistically analyzing the user evaluation results. For example, in Application Example 2, the volunteers generally reported that after using the essence containing silk fibroin aggregate-based dispersed organic active substances, their skin felt more moisturized, absorbed faster, and their skin condition was better after use. This verifies from a sensory perspective that the silk fibroin aggregate-based dispersed organic active substances enhance the effect of the essence.

[0136] Application Example 1

[0137] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 1 was added to the emulsion (the components of the emulsion include 6% glycerol, 89% water, 2% ethylene glycol, 2% butylene glycol and 1% cetyl ethylhexanoate in terms of mass fraction, and stirred and mixed evenly after addition) in an amount of 2 wt%. At the same time, vitamin C was added to the emulsion in an amount of 2 wt% as a comparison.

[0138] After testing, the emulsion has good stability and moisturizing effect, and the bioavailability of vitamin C is increased by 30%, while the bioavailability of vitamin C without silk fibroin treatment is only 20%, indicating that silk fibroin treatment can significantly improve the bioavailability of vitamin C.

[0139] Application Example 2

[0140] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 2 was added to the essence (the components of the essence include 90% water, 7% butylene glycol, 1% cetyl ethylhexanoate, and 2% tocopheryl acetate in terms of mass fraction) in an amount of 3wt%. At the same time, hyaluronic acid was added to the essence in an amount of 3wt% as a comparison.

[0141] After testing, the essence has good stability and moisturizing effect, and the bioavailability of hyaluronic acid is increased by 40%. In comparison, the bioavailability of hyaluronic acid in the essence that has not been treated with silk fibroin is about 30%, indicating that the addition of silk fibroin effectively improves the bioavailability of hyaluronic acid.

[0142] Application Example 3

[0143] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 3 was added to a facial cream (the components of the facial cream included 84% water, 5% polyethylene glycol, 5% propylene glycol and 6% sorbitol by mass) in an amount of 4 wt%. At the same time, coenzyme Q10 was added to the facial cream in an amount of 4 wt% as a comparison.

[0144] After testing, the cream has good stability and moisturizing effects, and the bioavailability of coenzyme Q10 is increased by 35%, while the bioavailability of untreated coenzyme Q10 is generally around 25%, indicating that silk fibroin aggregates can effectively improve the bioavailability of coenzyme Q10 in the cream.

[0145] Application Example 4

[0146] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 4 was added to a facial mask (the components of the facial mask include 85% water, 8% glycerin, 6% propylene glycol, 1% collagen and non-woven fabric in terms of mass fraction, and the non-woven fabric was added by soaking it in a solution of the above ingredients) in an amount of 5wt%. At the same time, niacinamide was added to the facial mask in an amount of 5wt% as a comparison.

[0147] After testing, the mask has good stability and moisturizing effects, and the bioavailability of niacinamide is increased by 45%; the bioavailability of niacinamide in the mask that has not been treated with silk fibroin is about 20%, indicating that niacinamide treated with silk fibroin shows better bioavailability in the mask.

[0148] Application Example 5

[0149] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 5 was added to the spray (the components of the spray included 78% water, 15% alcohol, 1% acrylic acid and 6% propylene glycol by mass fraction) in an amount of 1 wt %; at the same time, arbutin was added to the spray in an amount of 1 wt % as a comparison.

[0150] After testing, the spray has good stability and moisturizing effect, and the bioavailability of arbutin is increased by 32%; while the bioavailability of arbutin in the spray that has not been treated with silk fibroin is about 20%, indicating that the addition of silk fibroin significantly improves the bioavailability of arbutin.

[0151] Application Example 6

[0152] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 6 was added to the foundation (the components of the foundation include 73% water, 13% silicone oil, 5% polysorbate and 9% propylene glycol in terms of mass percentage) in an amount of 2 wt%. At the same time, kojic acid was added to the foundation in an amount of 2 wt% as a comparison.

[0153] After testing, the foundation has good stability and concealing effect, and the bioavailability of kojic acid is increased by 38%; the bioavailability of untreated kojic acid is about 30%, which shows that the bioavailability of kojic acid treated with silk fibroin in the foundation is higher.

[0154] Application Example 7

[0155] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 7 was added to lipstick (the components of lipstick include 48% beeswax, 45% paraffin, 2% cetyl ethylhexanoate and 5% octyldodecanol in terms of mass percentage) in an amount of 3 wt%. At the same time, licorice extract (licorice extract purchased from Jiangsu Caiwei Biotechnology Co., Ltd.) was added to the spray in an amount of 3 wt% as a comparison.

[0156] Testing showed that the lipstick had good stability and moisturizing effects, and the bioavailability of the licorice extract increased by 42%. The bioavailability of the licorice extract in the lipstick without silk fibroin treatment was about 30%, indicating that the addition of silk fibroin effectively improved the bioavailability of the licorice extract.

[0157] Application Example 8

[0158] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 8 was added to eye shadow (the components of the eye shadow include 60% beeswax, 20% water, 10% silicone oil and 10% talc in terms of mass percentage) in an amount of 4wt%. At the same time, Centella asiatica extract (Centella asiatica extract was purchased from Nanjing Recovery Biotechnology Co., Ltd.) was added to the spray in an amount of 4wt% as a comparison.

[0159] After testing, the eye shadow has good stability and color development effect, and the bioavailability of Centella asiatica extract is increased by 48%, while the bioavailability of untreated Centella asiatica extract is about 35%, indicating that the bioavailability of Centella asiatica extract treated with silk fibroin in eye shadow is higher.

[0160] Application Example 9

[0161] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 9 was added to the blush in an amount of 5 wt %; at the same time, chamomile extract (purchased from Fufeng Snot Biotechnology Co., Ltd.) was added to the blush in an amount of 5 wt % as a comparison.

[0162] After testing, the blush has good stability and color development effect, and the bioavailability of chamomile extract is increased by 50%. The bioavailability of chamomile extract not treated with silk fibroin in blush is about 30%, indicating that the addition of silk fibroin significantly improves the bioavailability of chamomile extract.

[0163] Application Example 10

[0164] The silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 10 was added to hand cream (the components of the hand cream include 62% beeswax, 21% glycerol, 10% propylene glycol and 7% olive oil in terms of mass percentage) in an amount of 3wt%. At the same time, green tea extract (purchased from Wuxi Sun Green Treasure Technology Co., Ltd.) was added to blush in an amount of 3wt% as a comparison.

[0165] Testing showed that the hand cream exhibited good stability and moisturizing effects, and the bioavailability of the green tea extract was increased by 40%. The bioavailability of untreated green tea extract in regular hand creams is approximately 26%. The green tea extract treated with silk fibroin exhibited even higher bioavailability in the hand cream.

[0166] Comparative Application Example 1

[0167] The gelatin-based dispersed organic active substance prepared in Comparative Example 1 was added to the emulsion (the components of the emulsion include 6% glycerol, 89% water, 2% ethylene glycol, 2% butylene glycol and 1% cetyl ethylhexanoate in terms of mass percentage, and stirred and mixed evenly after addition) in an amount of 2 wt%. After testing, it was found that when gelatin was used to treat organic active substances, although gelatin had certain film-forming and protective effects, due to its relatively simple molecular structure, its encapsulation ability for organic active substances was weak, and it was difficult to form a stable complex. The bioavailability of vitamin C in cosmetics after gelatin treatment was only increased by about 10%, which was much lower than the improvement in bioavailability of vitamin C in cosmetics after treatment with the silk fibroin aggregate-based dispersed organic active substance prepared in Preparation Example 1; and the stability improvement of the organic active substance treated with gelatin was limited (in the centrifugal stability test, the gelatin-treated emulsion showed slight stratification after centrifugation at 3000rpm for 30 minutes. In the long-term storage stability test, the gelatin-treated emulsion showed thinning and color change after storage at 40°C for 3 months. In the thermal stability test, the gelatin-treated emulsion showed thinning after being placed in a high temperature environment of 50°C for 1 week and then cooled to room temperature).

[0168] Application Comparative Example 2

[0169] The collagen-based dispersed organic active substance prepared in Comparative Example 2 is added to an essence (the components of the essence include 90% water, 7% butylene glycol, 1% cetyl ethylhexanoate and 2% tocopheryl acetate according to mass fraction), and the addition amount is 3wt%; Tests show that when collagen is used for treatment, although collagen has good biocompatibility, due to its large molecular weight, poor solubility and dispersibility, it is not tightly combined with the organic active substance, and the preparation process is complicated and difficult to disperse evenly. Taking hyaluronic acid as an example, the bioavailability of hyaluronic acid in cosmetics after collagen treatment is only increased by about 15% (the hyaluronic acid treated with silk fibroin aggregate base increases by 40%), and the stability in the formula is still not ideal.

[0170] From the above, it can be seen that compared with gelatin and collagen, silk fibroin has a unique molecular structure and excellent performance. Its molecular structure contains a large number of repetitive sequences and rich amino acid composition, which can form a close interaction with organic active substances, effectively improving the dispersibility and stability of organic active substances. Silk fibroin has good solubility and dispersibility. Through the preparation method of the present invention, it can be evenly mixed with organic active substances to form a stable complex. For example, in Example 1, the bioavailability of vitamin C in the emulsion after silk fibroin treatment was increased by 30%, which is significantly higher than the effect of gelatin and collagen treatment. In addition, silk fibroin also has good biocompatibility and moisturizing properties, which can further enhance the use effect and user experience of cosmetics. In terms of stability, silk fibroin aggregate-based dispersed organic active substances show good stability in different cosmetic formulas, are not easily affected by environmental factors, and effectively solve the stability problem of organic active substances in cosmetics.

[0171] In summary, silk fibroin has significant advantages in improving the bioavailability and stability of organic active substances, is superior to other types of protein processing methods, and can better meet the cosmetics industry's demand for the application of organic active substances.

[0172] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should all be covered by the scope of the claims of the present invention.

Claims

1. A method for preparing a silk fibroin aggregate-based dispersed organic active substance, characterized in that: The preparation method comprises: (1) dissolving an organic active substance in a first solvent to obtain a first solution; and dissolving a silk fibroin aggregate in a second solvent to obtain a second solution; (2) slowly adding the second solution into the first solution and mixing them to homogenize the silk fibroin aggregate-based dispersed organic active substance; The silk fibroin aggregate is prepared by mixing a silk fibroin aqueous solution with alcohol.

2. The preparation method according to claim 1, characterized in that The organic active substance is selected from one or more combinations of vitamin C, vitamin E, hyaluronic acid, coenzyme Q10, ceramide, niacinamide, arbutin, kojic acid, licorice extract, centella asiatica extract, chamomile extract, green tea extract, grape seed extract, squalane, tocopherol, retinol, retinol derivatives, peptide active substances, phytosterols or tetrahydromagnanol; and / or The first solvent and the second solvent are independently selected from one or more combinations of water, ethanol, propylene glycol or glycerol.

3. The preparation method according to claim 1 or 2, characterized in that The preparation method meets one or more of the following conditions: (i) In step (2), stirring is performed during mixing at a speed of 100 rpm to 1000 rpm for a time of 10 min to 60 min; (ii) In step (2), the homogenization pressure is 100 bar to 500 bar, and the homogenization time is 5 min to 30 min.

4. The silk fibroin aggregate-based dispersed organic active substance prepared by the preparation method according to any one of claims 1 to 3.

5. A cosmetic, characterized in that The invention comprises the silk fibroin aggregate-based dispersed organic active substance as described in claim 4.

6. The cosmetic according to claim 5, characterized in that Cosmetics include lotion, serum, cream, mask, spray, foundation, lipstick, eye shadow or blush.

7. The cosmetic according to claim 5 or 6, characterized in that The mass fraction of the silk fibroin aggregate-based dispersed organic active substance in the cosmetics is 0.1wt%-10wt%.

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