Collagen bionic inclusion as well as preparation method and application thereof

Abstract

The invention relates to a bionic inclusion of collagen as well as a preparation method and application of the bionic inclusion. The bionic inclusion comprises an oil phase, a water phase and collagen, the oil phase is prepared from ceramide, cholesterol, 7-dehydrocholesterol, fatty acid and grease; the water phase comprises an emulsifier and water. According to the bionic inclusion provided by the invention, oil phase components are reasonably compounded, so that the cuticle penetrating power and corium layer targeting property of collagen can be remarkably improved, and the long-acting targeted retention of the collagen on the corium layer is promoted.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of cosmetic technology, in particular to a biomimetic collagen package and a preparation method and application thereof. BACKGROUND

[0002] Collagen cosmetics have certain effects on human skin. They can form a moisturizing film on the skin surface, reduce water loss, and improve dryness. They can also temporarily support the skin, increase elasticity, and fade fine lines. However, there are obvious limitations. Collagen is a macromolecular protein, which is difficult to absorb through the skin due to the influence of the stratum corneum barrier, and it is difficult to penetrate the dermis to play a role. It cannot truly supplement the lost collagen in the skin, and the effect is mostly superficial and temporary. In the prior art, collagen is usually added directly in the form of an aqueous solution or wrapped by liposomes, microemulsions, etc. to improve stability and permeability.

[0003] CN118384055A discloses a high-permeability recombinant collagen liposome and a preparation method and application thereof. Type I recombinant collagen, type III recombinant collagen, and type V / XIV recombinant collagen are added to a supramolecular mixture composed of betaine and xylitol to form a first collagen mixture. The first collagen mixture is freeze-dried to obtain a collagen-supramolecular sponge, which is then dissolved in water to obtain a second collagen mixture. Sodium (dodecylamide glutamate) lysine and lecithin are mixed to obtain a first mixture. The second collagen mixture is added to the first mixture under water bath and continuous stirring conditions, followed by homogenization to obtain a recombinant collagen liposome, thereby improving its stability and permeability.

[0004] A document ("Transdermal transport of collagen and hyaluronic acid using water in oil microemulsion", Patrycja et al., International Journal of Pharmaceutics, Vol. 572) studies a collagen transdermal delivery based on water-in-oil microemulsion. A surfactant mixture and an oil phase are mixed. Collagen and hyaluronic acid are dissolved in an aqueous phase containing propylene glycol and ethanol. A stable microemulsion is formed by high-speed homogenization. By wrapping collagen in a microemulsion, its permeability is improved.

[0005] Although the above-mentioned technologies improve the permeability of collagen to some extent, there are still the following problems: 1) liposome technology: the preparation process is complex, needs to be freeze-dried, and is difficult to scale up; the stability of the liposome is poor, and degradation or leakage easily occurs, resulting in invalidation of the collagen. 2) microemulsion technology: a large amount of surfactant and co-surfactant is needed, which may cause irritation to the skin; the stability of the microemulsion system is greatly affected by factors such as temperature and pH value, and the applicable range is limited. 3) direct water solution addition: the transdermal absorption rate is low, the collagen is easily degraded, and the skin care effect is limited.

[0006] Therefore, it is necessary to provide a biomimetic package of collagen to solve the problems of short retention time, shallow action site and complex preparation process of collagen in the prior art, and significantly improve the targeted release and efficacy of collagen. SUMMARY

[0007] To solve the above technical problems, the present application provides a biomimetic package of collagen, its preparation method and application. The biomimetic package provided by the present application can significantly improve the stratum corneum penetration and dermal targeting of collagen by reasonable compounding of oil phase components, and promote long-acting targeted retention of collagen in the dermis.

[0008] To achieve this purpose, the present application adopts the following technical solutions:

[0009] In a first aspect, the present application provides a biomimetic package of collagen, which comprises an oil phase, an aqueous phase and collagen; the oil phase comprises ceramide, cholesterols, 7-dehydrocholesterol, fatty acids and oil; the aqueous phase comprises an emulsifier and water.

[0010] In the present application, the omega-hydroxy fatty acid chain of ceramide forms a directional water molecule chain (each hydroxyl group can bind 3-5 water molecules), which improves the hydration degree of the stratum corneum; the rigid sterol nucleus of cholesterols inserts into the lipid layer, generating local curvature stress; 7-dehydrocholesterol is specifically synthesized by the epidermal cells of the skin, which can form intermolecular hydrogen bonds with the hydroxyl groups of ceramide, reducing the liquid crystal phase transition temperature from 42℃ to 35℃, adapting to the skin surface temperature (32±2℃), and playing a long-acting slow-release role; the cis-double bond of fatty acids forms a "molecular hinge" structure, which forms a "fluid microzone" with cholesterols, destroys the close packing of stratum corneum lipids, increases the fluidity of the lipid bilayer, and reduces the diffusion resistance; the oil acts as a solvent.

[0011] The present application provides a biomimetic package for collagen, which is difficult to absorb transdermally as a macromolecular protein. By reasonable compounding of lipid molecules in the oil phase, the skin lipid composition is accurately simulated, and a new transdermal mechanism is formed, which can significantly improve the stratum corneum penetration and dermal retention of collagen.

[0012] Preferably, the oil phase comprises ceramide 1-10 parts by mass, cholestanol 1-10 parts by mass, 7-dehydrocholestanol 1-10 parts by mass, fatty acid 1-10 parts by mass, and oil 10-30 parts by mass.

[0013] In the present application, the amount of ceramide can be, for example, 1 part, 3 parts, 5 parts, 6 parts, 9 parts, 10 parts, etc.; the amount of cholestanol can be, for example, 1 part, 2 parts, 3 parts, 5 parts, 7 parts, 9 parts, 10 parts, etc.; the amount of 7-dehydrocholestanol can be, for example, 1 part, 2.5 parts, 4 parts, 6 parts, 8.5 parts, 10 parts, etc.; the amount of fatty acid can be, for example, 1 part, 1.2 parts, 3.8 parts, 4.4 parts, 7.3 parts, 8.6 parts, 9.9 parts, 10 parts, etc.; and the amount of oil can be, for example, 10 parts, 13 parts, 18 parts, 21 parts, 26 parts, 30 parts, etc.

[0014] Preferably, the fatty acid comprises any one or a combination of at least two of palmitic acid, stearic acid, or oleic acid.

[0015] Preferably, the oil comprises any one or a combination of at least two of caprylic / capric triglyceride, dimethicone, soybean oil, isononyl isononanoate, rice bran oil, mineral oil, sunflower seed oil, glyceryl tri(ethylhexyl) citrate, ethylhexyl palmitate, isododecane, or squalane.

[0016] Preferably, the oil phase further comprises C14-C22 fatty alcohol.

[0017] Preferably, the oil phase further comprises C14-C22 fatty alcohol 1-10 parts by mass (for example, 1 part, 1.8 parts, 2.8 parts, 4 parts, 5.2 parts, 7.3 parts, 10 parts, etc.).

[0018] Preferably, the C14-C22 fatty alcohol comprises any one or a combination of at least two of cetearyl alcohol, cetyl alcohol, stearyl alcohol, arachidyl alcohol, behenyl alcohol, batyl alcohol, or myristyl alcohol.

[0019] In the present application, the straight-chain alkyl group of the C14-C22 fatty alcohol matches the carbon chain length of the ceramide in the stratum corneum of the skin, and can form a regular lamellar liquid crystal structure, simulating the natural lipid barrier arrangement pattern.

[0020] Preferably, the water phase comprises emulsifier 1-5 parts by mass (for example, 1 part, 2.3 parts, 3.7 parts, 4 parts, 4.3 parts, 5 parts, etc.) and water 70-150 parts by mass (for example, 70 parts, 88 parts, 97 parts, 111 parts, 136 parts, 150 parts, etc.).

[0021] Preferably, the emulsifier comprises any one of hydrogenated lecithin, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, or polyoxyethylene sorbitan monooleate, or a combination of at least two thereof.

[0022] Preferably, the mass ratio of the oil phase, the water phase, and the collagen is (15-65):(70-150):(0.01-1).

[0023] The above numerical value (15-65) may be, for example, 15, 25, 35, 45, 55, 65, etc.; (70-150) may be, for example, 70, 80, 100, 120, 140, 150, etc.; and (0.01-1) may be, for example, 0.01, 0.1, 0.2, 0.5, 0.8, 1, etc.

[0024] Preferably, the biomimetic inclusion body further comprises a preservative.

[0025] Preferably, the preservative comprises any one of phenoxyethanol, octisalate, or ethylhexylglycerin, or a combination of at least two thereof.

[0026] In a second aspect, the present application provides a preparation method of the biomimetic inclusion body of collagen according to the first aspect, the preparation method comprising the following steps:

[0027] (1) mixing raw materials of the oil phase, heating and dissolving to obtain the oil phase;

[0028] (2) mixing raw materials of the water phase, heating and dissolving to obtain the water phase;

[0029] (3) mixing the oil phase and the water phase, and then homogenizing to obtain the biomimetic penetration system;

[0030] (4) mixing the biomimetic penetration system and the collagen to obtain the biomimetic inclusion body of collagen;

[0031] Wherein, the order of steps (1) and (2) is not limited.

[0032] Preferably, the heating temperature of step (1) is 100-120℃ (for example, it may be 100℃, 105℃, 110℃, 115℃, 120℃, etc.).

[0033] Preferably, the heating temperature of step (2) is 80-90℃ (for example, it may be 80℃, 82℃, 85℃, 88℃, 90℃, etc.).

[0034] Preferably, the mixing of step (3) is: pouring the water phase into the oil phase.

[0035] In the present application, the water phase is quickly poured into the oil phase, a high shear force instantaneously generates a huge phase interface area, forcing the lipid molecules (such as ceramide, cholesterols) to orient and arrange at the interface, forming a layered liquid crystal precursor similar to the skin lipid layer; it can also avoid the excessive growth of ceramide beta-fold crystallization, ensure the liquid crystal layer spacing formed close to the natural stratum corneum structure, thereby improving the transdermal absorption effect.

[0036] Preferably, the temperature of the mixing in step (3) is 85-95℃ (for example, it can be 85℃, 88℃, 90℃, 92℃, 95℃, etc.).

[0037] Preferably, the temperature of the homogenization in step (3) is 80-90℃ (for example, it can be 80℃, 82℃, 85℃, 88℃, 90℃, etc.), the rotation speed is 5000-7000rpm (for example, it can be 5000rpm, 5500rpm, 6000rpm, 6500rpm, 7000rpm, etc.), and the time is 1-2min (for example, it can be 1min, 1.2min, 1.5min, 1.8min, 2min, etc.).

[0038] In the present application, by controlling the preparation temperature of the oil phase and the water phase, the mixing temperature of the two, and the homogenization temperature, the lipid is always kept in a molten state, which is beneficial to co-assembly.

[0039] Preferably, the temperature of the mixing in step (4) is 30-40℃ (for example, it can be 30℃, 32℃, 35℃, 38℃, 40℃, etc.).

[0040] Preferably, the mixing in step (4) further includes adding a preservative.

[0041] In a third aspect, the present application provides a use of the biomimetic niosome of collagen according to the first aspect in the preparation of a cosmetic product with long-acting targeted retention effect of collagen.

[0042] Compared with the prior art, the present application has at least the following beneficial effects:

[0043] (1) The oil phase of the biomimetic niosome provided by the present application simulates the skin lipid composition by compounding lipid molecules, effectively improves the stratum corneum penetration and dermis retention of collagen, and can realize the effects of intelligent controlled release and long-acting retention in the dermis.

[0044] (2) The biomimetic niosome provided by the present application uses skin-specific constituent lipids, is mild and non-irritating to the skin, has a simple preparation method, is easy to scale up, can be applied to various anti-aging serums, creams, and medical dressings, etc. cosmetic categories, and has a wide application prospect. DETAILED DESCRIPTION

[0045] For the purpose of understanding the present application, the present application is illustrated by the following examples. It should be apparent to those skilled in the art that the examples are merely illustrative of the present application and should not be viewed as limiting the present application.

[0046] Example 1-3

[0047] Example 1-3 respectively provides a biomimetic package of collagen, the component types and mass fractions of which are shown in the following table:

[0048]

[0049]

[0050] The preparation method of the biomimetic package of collagen comprises the following steps:

[0051] (1) Mix the raw materials of the oil phase, heat and dissolve at 110℃ to obtain the oil phase; mix the raw materials of the water phase, heat and dissolve at 80℃ to obtain the water phase;

[0052] (3) Pour the water phase into the oil phase quickly at 90℃ to form a water-oil mixture, and then homogenize at 90℃ and 6000rpm for 2min to obtain a biomimetic penetration system;

[0053] (4) When the biomimetic penetration system is cooled to 35℃, add the collagen aqueous solution and the preservative under stirring (80rpm) to obtain the biomimetic package of collagen;

[0054] Example 4

[0055] This example provides a biomimetic package of collagen, which is different from Example 1 only in that C14-C22 fatty alcohol is not added, and its mass fraction is proportionally distributed to other components in the oil phase, and the rest is referred to Example 1.

[0056] Example 5

[0057] This example provides a biomimetic package of collagen, which is different from Example 1 only in that in the preparation method step (3), the oil phase is poured into the water phase quickly at 90℃, and the rest is referred to Example 1.

[0058] Comparative Example 1

[0059] This comparative example provides a biomimetic package of collagen, which is different from Example 1 only in that ceramide NP is not added, and its mass fraction is proportionally distributed to cholesterols, 7-dehydrocholesterol and fatty acids, and the rest is referred to Example 1.

[0060] Comparative Example 2

[0061] This comparative example provides a biomimetic collagen wrapping body, which is only different from Example 1 in that no cholesterols are added, and the mass fraction thereof is proportionally distributed to ceramide NPs, 7-dehydrocholesterol and fatty acids, and the rest refers to Example 1.

[0062] Comparative Example 3

[0063] This comparative example provides a biomimetic collagen wrapping body, which is only different from Example 1 in that no 7-dehydrocholesterol is added, and the mass fraction thereof is proportionally distributed to cholesterols, ceramide NPs and fatty acids, and the rest refers to Example 1.

[0064] Comparative Example 4

[0065] This comparative example provides a biomimetic collagen wrapping body, which is only different from Example 1 in that no fatty acids are added, and the mass fraction thereof is proportionally distributed to cholesterols, 7-dehydrocholesterol and ceramide NPs, and the rest refers to Example 1.

[0066] Test Example

[0067] In-vitro transdermal penetration experiment (intracutaneous retention amount determination)

[0068] Test Method:

[0069] The transdermal experiment of isolated pigskin was carried out by vertical Franz diffusion cell method. The pigskin was fixed in the receiving chamber and the supply chamber, and 0.5g of the sample prepared in each example and comparative example was taken in the supply chamber, and PBS buffer solution was used as the receiving liquid, and the diffusion was stirred at 32°C.

[0070] After 12h and 24h of transdermal penetration, the pigskin was taken out of the diffusion cell, the residual sample on the surface of the pigskin was wiped clean with absorbent cotton, and then the content of collagen in the stratum corneum was determined by tape stripping method, 21 layers of skin on the surface of the pigskin were adhered with 3M tape, the first layer was discarded, and the remaining 20 layers were placed in a 15mL centrifuge tube, 2mL of PBS buffer solution was added to the tube, ultrasonic treatment was carried out for 30min, centrifugation was carried out at 5000rpm for 30min, the supernatant was filtered with a 0.22μm organic filter membrane, and the content of collagen was determined by HPLC method, which was the content of collagen in the stratum corneum.

[0071] The remaining skin surface was heated at 60°C for 15s, and the viable epidermis layer was carefully cut off with a scalpel, and then cut into pieces and placed in a 15mL centrifuge tube, 2mL of PBS buffer solution was added, ultrasonic treatment was carried out for 30min, centrifugation was carried out at 5000rpm for 30min, the supernatant was filtered with a 0.22μm filter membrane, and the content of collagen was determined by HPLC method, which was the content of collagen in the viable epidermis layer.

[0072] The last remaining skin was cut into pieces and placed in a 15 mL centrifuge tube, 2 mL PBS buffer solution was added, ultrasonic treatment for 30 min, then centrifuged at 5000 rpm for 30 min, the supernatant was filtered by 0.22 μm organic filter membrane, and the content of collagen was determined by HPLC method, which was the content of collagen in the dermis layer.

[0073] The biomimetic inclusion bodies provided by Examples 1-5 and Comparative Examples 1-4 were tested, and the results are shown in Table 1.

[0074] Table 1

[0075]

[0076] According to the data in Table 1, the 12h, 24h stratum corneum collagen retention rate and 12h, 24h dermis collagen targeting retention rate were calculated, and the results are shown in Table 2.

[0077] Stratum corneum collagen retention rate = Stratum corneum collagen content / (Stratum corneum collagen content + Epidermis collagen content + Dermis collagen content)

[0078] Dermis collagen targeting retention rate = Dermis collagen content / (Stratum corneum collagen content + Epidermis collagen content + Dermis collagen content)

[0079] Table 2

[0080]

[0081] Test results:

[0082] (1) As can be seen from Examples 1 to 5, the present application can significantly improve the retention time and penetration depth of collagen by reasonable compounding of the oil phase components in the biomimetic inclusion body of collagen, improve the stratum corneum penetration, and promote the long-term targeted retention of collagen in the dermis.

[0083] (2) As can be seen from the comparison of Example 1 and Example 5, in the preparation method of the present application, the water phase is quickly poured into the oil phase, which is beneficial to the directional arrangement of lipid molecules in the oil phase at the interface, forming a lamellar liquid crystal precursor similar to the skin lipid layer; It can also avoid the excessive growth of ceramide beta-folded crystal, ensure that the liquid crystal layer spacing is close to the natural stratum corneum structure, and achieve better technical effect of improving the retention time and penetration depth of collagen.

[0084] (3) As can be seen from the comparison of Example 1 and Comparative Examples 1-4, when any one of ceramide, cholesterols, 7-dehydrocholesterol and fatty acids is missing in the oil phase of the biomimetic inclusion body, the stratum corneum penetration of collagen and the targeting of the dermis are significantly reduced.

[0085] The applicant states that the above description is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto. It should be understood by those skilled in the art that any changes or replacements within the technical scope disclosed by the present application can be easily conceived by any person skilled in the art, and all such changes and replacements fall within the protection scope and disclosure scope of the present application.

Claims

1. A biomimetic collagen inclusion, characterized in that: The bionic inclusion comprises an oil phase, a water phase and collagen; the oil phase comprises ceramide, cholesterol, 7-dehydrocholesterol, fatty acid and oil; and the water phase comprises an emulsifier and water.

2. The bionic inclusion according to claim 1, characterized in that: The oil phase comprises, by mass, 1-10 parts of ceramide, 1-10 parts of cholesterol, 1-10 parts of 7-dehydrocholesterol, 1-10 parts of fatty acid and 10-30 parts of oil.

3. The bionic inclusion according to claim 1 or 2, characterized in that: The fatty acid includes any one of palmitic acid, stearic acid or oleic acid, or a combination of at least two thereof; Preferably, the oil comprises any one or a combination of at least two of caprylic / capric triglyceride, dimethicone, soybean oil, isononyl isononanoate, rice bran oil, mineral oil, sunflower seed oil, triethylhexyl glyceryl, ethylhexyl palmitate, isododecane or squalane.

4. The bionic inclusion according to any one of claims 1 to 3, characterized in that: The oil phase also includes C14-C22 fatty alcohol; Preferably, the oil phase further comprises 1-10 parts by mass of C14-C22 fatty alcohol; Preferably, the C14-C22 fatty alcohol includes any one of cetearyl alcohol, cetyl alcohol, stearyl alcohol, arachidyl alcohol, behenyl alcohol, batyl alcohol or myristyl alcohol, or a combination of at least two thereof.

5. The bionic inclusion according to any one of claims 1 to 4, characterized in that: The aqueous phase comprises 1-5 parts of emulsifier and 70-150 parts of water by mass; Preferably, the emulsifier includes any one of hydrogenated lecithin, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate or polyoxyethylene sorbitan monooleate, or a combination of at least two thereof.

6. The bionic inclusion according to any one of claims 1 to 5, characterized in that: The mass ratio of the oil phase, the water phase and the collagen is (15-65):(70-150):(0.01-1).

7. A method for preparing a biomimetic collagen inclusion according to any one of claims 1 to 6, characterized in that: The preparation method comprises the following steps: (1) mixing the raw materials of the oil phase, heating and dissolving them to obtain the oil phase; (2) mixing the raw materials of the aqueous phase, heating and dissolving them to obtain an aqueous phase; (3) mixing the oil phase and the water phase and then homogenizing to obtain a biomimetic penetration-enhancing system; (4) mixing the bionic penetration-enhancing system and collagen to obtain a bionic inclusion of the collagen; The order of steps (1) and (2) is irrelevant.

8. The preparation method according to claim 7, characterized in that The heating temperature in step (1) is 100-120° C. Preferably, the heating temperature in step (2) is 80-90°C.

9. The preparation method according to claim 7 or 8, characterized in that The mixing in step (3) is as follows: pouring the water phase into the oil phase; Preferably, the mixing temperature in step (3) is 85-95°C; Preferably, the homogenization temperature in step (3) is 80-90°C, the speed is 5000-7000 rpm, and the time is 1-2 min; Preferably, the mixing temperature in step (4) is 30-40°C.

10. Use of the biomimetic inclusion of collagen according to any one of claims 1 to 6 in preparing cosmetics having a long-lasting targeted retention effect of collagen.

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