Composite vitamin derivative nano-liposome and preparation method thereof

Abstract

The invention provides a composite vitamin derivative nano-liposome and a preparation method thereof, and relates to the technical field of nano-liposomes. Every 100 parts by weight of the nano-liposome comprises the following raw materials: 15-30 parts of polyol, 3-15 parts of a vitamin complex derivative, 1-3 parts of lecithin, 0.5-5 parts of a softening and moistening agent, 5-10 parts of an emulsifier, 0.5-2 parts of a preservative and the balance of purified water. According to the method provided by the invention, the hydrophilicity of three indissolvable vitamin derivatives is improved, and the solubility of the three indissolvable vitamin derivatives in an aqueous matrix is improved; meanwhile, according to the liposome provided by the invention, the components are reasonably proportioned, so that synergistic interaction is realized, and the anti-oxidation, anti-whitening and anti-oxidation effects are enhanced; the liposome has excellent dispersity, stability and biocompatibility, active ingredients are effectively wrapped, direct stimulation of the active ingredients to the skin is avoided, the safety of the product is improved, and meanwhile, the liposome can achieve the effect that the active ingredients directly reach the bottom of the skin through the surface layer of the skin to act on a target spot position.

Description

Technical Field

[0001] The invention relates to the technical field of nano liposomes, and in particular to a composite vitamin derivative nano liposome and a preparation method thereof. Background Art

[0002] In recent years, a skin care concept of "C in the morning and A in the evening" has emerged in the skin care industry. This skin care concept refers to using skin care products containing vitamin C in the morning, because you will be exposed to more ultraviolet rays during the day, and ultraviolet rays will accelerate the production of free radicals in the body, so using vitamin C products in the morning can help with anti-oxidation. Use skin care products containing vitamin A at night, because vitamin A is sensitive to light and can easily cause skin irritation and allergic reactions. Therefore, using retinol at night can avoid it from being affected by sunlight and reduce skin discomfort. This concept aims to scientifically identify skin needs in different time periods, use appropriate skin care products at the right time, and maximize the skin care effect. However, for modern fast-paced life, skin care seems to have become very "troublesome", and you have to spend energy to choose different products and distinguish the time of use.

[0003] Nowadays, consumers are increasingly advocating the concept of "streamlined skin care". It is not a good idea to directly "pile" things on your face. First of all, the irritation of active ingredients, such as vitamin A products, may make the skin condition worse if tolerance is not established or improper usage habits are used. Secondly, many active ingredients cannot penetrate into the bottom layer of the skin due to their large molecular weight. They are metabolized after staying on the surface of the skin for a period of time. Finally, different products use different formula systems and different excipients. Will there be more risks of irritation and burden on the skin when used in the morning and evening? Summary of the invention

[0004] 1. Technical issues to be resolved

[0005] In view of the deficiencies of the prior art, the present invention provides a composite vitamin derivative nanoliposome and a preparation method thereof.

[0006] (II) Technical solution

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] The invention provides a composite vitamin derivative nano liposome. The raw materials of the nano liposome, per 100 parts by weight, include the following raw materials: 15-30 parts of polyol, 3-15 parts of composite vitamin derivative, 1-3 parts of lecithin, 0.5-5 parts of softener, 5-10 parts of emulsifier, 0.5-2 parts of preservative, and the rest is purified water.

[0009] Preferably, every 100 parts by weight of the raw materials of the nano-liposomes include the following raw materials: 20-30 parts of polyol, 6-15 parts of compound vitamin derivative, 1-2 parts of lecithin, 0.5-3 parts of emollient, 5-8 parts of emulsifier, 0.5-1 part of preservative, and the balance being purified water.

[0010] Specifically, the compound vitamin derivative is composed of retinyl palmitate, ascorbyl tetraisopalmitate, and tocopheryl acetate at a weight ratio of 1-5:1-5:1-5.

[0011] Specifically, the compound vitamin derivative is composed of retinyl palmitate, ascorbyl tetraisopalmitate, and tocopheryl acetate at a weight ratio of 1:1:1.

[0012] Specifically, the polyol is one or more of 1,2-propanediol, 1,3-propanediol, 1,4-butanediol, 1,2-pentanediol, glycerol, and polyethylene glycol.

[0013] Specifically, the emulsifier is one or more of polysorbate-20, glyceryl stearate, sodium lauryl polyoxyethylene ether sulfate, sodium cetyl polyoxyethylene ether phosphate, C12-14 alkyl dimethyl benzyl ammonium chloride, cocamidopropyl betaine, and sorbitan monolaurate.

[0014] Specifically, the emollient is one or more of cholesterol, allantoin, hexyl decanol, octyldecanol, cetyl alcohol, and isopropyl palmitate.

[0015] Specifically, the lecithin is one or more of hydrogenated lecithin, soybean lecithin, and egg yolk lecithin.

[0016] Specifically, the preservative is one or more of phenoxyethanol, potassium sorbate, glyceryl caprate, capric acid, glyceryl caprylate, and sorbitan caprylate.

[0017] The present invention also provides a preparation method of compound vitamin derivative nano-liposomes. The nano-liposomes are prepared according to the following steps:

[0018] S1: Take 1-5 parts of the active ingredient retinyl palmitate, add 5-10 parts of polyol; stir at a normal temperature of 100-300 rpm for 10-30 min to obtain mixture 1.

[0019] S2: Take 1-5 parts of the active ingredient ascorbyl tetraisopalmitate, add 5-10 parts of polyol, stir at a normal temperature of 100-300 rpm for 10-30 min to obtain mixture 2;

[0020] S3: Take 1 - 5 parts of the active ingredient tocopheryl acetate, add 5 - 10 parts of polyol, place it at room temperature and stir at a speed of 100 - 300 rpm for 10 - 30 min to obtain mixture 3;

[0021] S4: Transfer mixture 1 - 3 into an oil - phase premixing system, heat it to 30 - 50 °C, and stir at 100 - 500 rpm for 10 - 30 min to obtain mixture A;

[0022] S5: Add 1 - 3 parts of lecithin and 0.5 - 5 parts of emollient to mixture A, raise the temperature to 60 - 65 °C, and continue to stir at 100 - 500 rpm for 10 - 30 min to obtain mixture B;

[0023] S6: Prepare 5 - 10 parts of emulsifier and 0.5 - 2 parts of preservative, add them to pure water, and stir and disperse in a water bath at 50 - 70 °C to obtain mixture C;

[0024] S7: Slowly add mixture B to mixture C, set the shear speed to 5000 - 8000 rpm; the shear temperature is 50 - 70 °C; the shear time is 3 - 8 min to prepare crude liposomes;

[0025] S8: Subject the crude liposomes to high - pressure homogenization treatment, with a homogenization pressure of 400 - 800 bar, a temperature of 50 - 65 °C; the number of cycles is 1 - 5 times, and cool at room temperature for at least 2 h to obtain the composite vitamin derivative liposomes.

[0026] (III) Beneficial Effects

[0027] The present invention provides a composite vitamin derivative nano - liposome and its preparation method. By using the method provided by the present invention, the hydrophilicity of three poorly soluble vitamin derivative active ingredients is improved, enabling the above - mentioned ingredients to enhance their solubility in the aqueous matrix; at the same time, the liposomes provided by the present invention rationally proportion the three active ingredients, enabling them to achieve synergistic effects and strengthening the effects of antioxidant, anti - oxidation, and anti - whitening; the liposomes obtained by the present invention are nanoscale, with excellent dispersibility and stability; the liposomes effectively encapsulate the active ingredients, not only avoiding the direct stimulation of the active ingredients to the skin and improving the safety of the product, but also enabling the active substances to penetrate through the skin surface and directly act on the target position at the bottom layer of the muscle, thereby improving the bioavailability of the active substances. Description of the Drawings

[0028] Figure 1 Retention rate and particle size change of samples in Examples 1 - 7

[0029] Figure 2 5% diluted aqueous solution of the sample in Example 1

[0030] Figure 3Inhibitory effects of the samples of Example 1 and Comparative Examples 1-8 on IL-1α

[0031] Figure 4 Inhibitory effects of the samples of Example 1 and Comparative Examples 1-8 on PGE2

[0032] Figure 5 DPPH free radical scavenging rates of the samples of Example 1 and Comparative Examples 1-8

[0033] Figure 6 ABTS free radical scavenging rates of the samples of Example 1 and Comparative Examples 1-8

[0034] Figure 7 Effects of the samples of Example 1 and Comparative Examples 1-8 on collagen synthesis

[0035] Figure 8 Anti-aging test on human skin Detailed implementation manners

[0036] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0037] Example 1

[0038] A composite vitamin derivative nano-liposome is prepared according to the following steps:

[0039] S1: Take 2 parts of the active ingredient retinol palmitate and add 8 parts of 1,2-propanediol; place it at room temperature and stir at 100 rpm for 20 min to obtain mixture 1;

[0040] S2: Take 2 parts of the active ingredient ascorbyl tetraisopalmitate and add 8 parts of 1,2-propanediol; place it at room temperature and stir at 100 rpm for 20 min to obtain mixture 2;

[0041] S3: Take 2 parts of the active ingredient tocopheryl acetate and add 8 parts of glycerol; place it at room temperature and stir at 100 rpm for 20 min to obtain mixture 3;

[0042] S4: Transfer mixtures 1-3 to an oil phase premixing system, heat to 40 °C, and stir at 100 rpm for 10 min to obtain mixture A;

[0043] S5: Add 2 parts of hydrogenated lecithin and 3 parts of hexyl decanol to the mixture A, raise the temperature to 60 °C, and stir at 100 rpm for 20 min to obtain the mixture B;

[0044] S6: Prepare 5 parts of glyceryl stearate and 0.5 part of potassium sorbate, add them to 59.5 parts of purified water, and stir and disperse in a water bath at 50 °C to obtain the mixture C;

[0045] S7: Under shear conditions, slowly add the mixture B to the mixture C, with a shear speed of 6000 rpm, a shear temperature of 50 °C, and a shear time of 5 min to prepare the crude liposomes;

[0046] S8: Subject the crude liposomes to high-pressure homogenization treatment, with a homogenization pressure of 500 bar and a temperature of 50 °C; the number of cycles is 2, and cool at room temperature for 2 h to obtain the complex vitamin derivative liposomes.

[0047] Example 2

[0048] A complex vitamin derivative nano-liposome is prepared according to the following steps:

[0049] S1: Take 3 parts of the active ingredient retinol palmitate, add 10 parts of glycerol; place it at room temperature and stir at 100 rpm for 30 min to obtain the mixture 1;

[0050] S2: Take 1.5 parts of the active ingredient ascorbyl tetraisopalmitate, add 6 parts of 1,2-pentanediol, place it at room temperature and stir at 300 rpm for 30 min to obtain the mixture 2;

[0051] S3: Take 1.5 parts of the active ingredient tocopheryl acetate, add 6 parts of 1,2-pentanediol, place it at room temperature and stir at 100 rpm for 10 min to obtain the mixture 3;

[0052] S4: Transfer the mixtures 1-3 into an oil phase premixing system, heat to 45 °C, and stir at 200 rpm for 30 min to obtain the mixture A;

[0053] S5: Add 1 part of soy lecithin and 2 parts of cholesterol to the mixture A, raise the temperature to 60 °C, and continue to stir at 100 rpm for 20 min to obtain the mixture B;

[0054] S6: Prepare 10 parts of polysorbate-20 and 1 part of glyceryl caprylate, add them to 58 parts of purified water, and stir and disperse in a water bath at 50 °C to obtain the mixture C;

[0055] S7: Under shear conditions, slowly add the mixture B to the mixture C, with a shear speed of 5000 rpm, a shear temperature of 60 °C, and a shear time of 3 min to prepare the crude liposomes;

[0056] S8: The crude liposomes are subjected to high-pressure homogenization treatment at a homogenization pressure of 500 bar, a temperature of 55 °C, and a circulation number of 3 times, and cooled at room temperature for 2 h to obtain the vitamin complex derivative liposomes.

[0057] Example 3

[0058] A vitamin complex derivative nano-liposome is prepared according to the following steps:

[0059] S1: Take 3.6 parts of the active ingredient retinol palmitate and add 10 parts of 1,3-propanediol; stir at a normal temperature of 200 rpm for 10 min to obtain mixture 1;

[0060] S2: Take 1.2 parts of the active ingredient ascorbyl tetraisopalmitate and add 5 parts of glycerol; stir at a normal temperature of 200 rpm for 10 min to obtain mixture 2;

[0061] S3: Take 1.2 parts of the active ingredient tocopheryl acetate and add 5 parts of glycerol; stir at a normal temperature of 100 rpm for 10 min to obtain mixture 3;

[0062] S4: Transfer mixtures 1 - 3 to an oil phase premixing system, heat to 30 °C, and stir at 300 rpm for 15 min to obtain mixture A;

[0063] S5: Add 3 parts of hydrogenated lecithin and 5 parts of isopropyl palmitate to mixture A, raise the temperature to 60 °C, and stir at 500 rpm for 10 min to obtain mixture B;

[0064] S6: Prepare 5 parts of cocamidopropyl betaine and 2 parts of sorbitan octanoate, add them to 59 parts of purified water, and stir and disperse in a 60 °C water bath to obtain mixture C;

[0065] S7: Under shear conditions, slowly add mixture B to mixture C at a shear speed of 5500 rpm, a shear temperature of 70 °C, and a shear time of 3 min to prepare the crude liposomes;

[0066] S8: The crude liposomes are subjected to high-pressure homogenization treatment at a homogenization pressure of 800 bar, a temperature of 60 °C, and a circulation number of 1 time, and cooled at room temperature for 2 h to obtain the vitamin complex derivative liposomes.

[0067] Example 4

[0068] A vitamin complex derivative nano-liposome is prepared according to the following steps:

[0069] S1: Take 1.5 parts of the active ingredient retinol palmitate and add 6 parts of glycerol; stir at a normal temperature of 200 rpm for 30 min to obtain mixture 1;

[0070] S2: Take 3 parts of the active ingredient ascorbyl tetraisopalmitate, add 10 parts of 1,2-pentanediol, place it at room temperature and stir at 200 rpm for 30 min to obtain mixture 2;

[0071] S3: Take 1.5 parts of the active ingredient tocopheryl acetate, add 6 parts of polyethylene glycol, place it at room temperature and stir at 200 rpm for 30 min to obtain mixture 3;

[0072] S4: Transfer mixtures 1 - 3 to an oil phase premixing system, heat to 40 °C, and stir at 300 rpm for 15 min to obtain mixture A;

[0073] S5: Add 1 part of egg yolk lecithin and 5 parts of cetyl alcohol to mixture A, raise the temperature to 65 °C, and continue to stir at 500 rpm for 10 min to obtain mixture B;

[0074] S6: Prepare 10 parts of sorbitan monolaurate, 2 parts of capric acid, add them to 54 parts of purified water, and stir and disperse in a 60 °C water bath to obtain mixture C;

[0075] S7: Slowly add mixture B to mixture C, with a shear speed of 6000 rpm, a shear temperature of 70 °C, and a shear time of 5 min to prepare crude liposomes;

[0076] S8: Subject the crude liposomes to high-pressure homogenization treatment, with a homogenization pressure of 600 bar, a temperature of 50 °C, and a circulation number of 1 time, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0077] Example 5

[0078] A composite vitamin derivative nano-liposome is prepared according to the following steps:

[0079] S1: Take 1.2 parts of the active ingredient retinol palmitate, add 5 parts of polyethylene glycol; place it at room temperature and stir at 300 rpm for 20 min to obtain mixture 1;

[0080] S2: Take 3.6 parts of the active ingredient ascorbyl tetraisopalmitate, add 10 parts of glycerol, place it at room temperature and stir at 300 rpm for 20 min to obtain mixture 2;

[0081] S3: Take 1.2 parts of the active ingredient tocopheryl acetate, add 5 parts of 1,2-pentanediol, place it at room temperature and stir at 300 rpm for 20 min to obtain mixture 3;

[0082] S4: Transfer mixtures 1 - 3 to an oil phase premixing system, heat to 50 °C, and stir at 200 rpm for 30 min to obtain mixture A;

[0083] S5: Add 2 parts of soy lecithin and 0.5 part of octyldodecanol to the mixed solution A, raise the temperature to 65 °C, and stir for 20 min at 300 rpm to obtain the mixed solution B;

[0084] S6: Prepare 5 parts of sodium lauryl polyoxyethylene ether sulfate and 2 parts of sorbitan octanoate, add them to 51.5 parts of purified water, and stir and disperse in a 70 °C water bath to obtain the mixed solution C;

[0085] S7: Slowly add the mixed solution B to the mixed solution C, with a shear speed of 6000 rpm, a shear temperature of 60 °C, and a shear time of 5 min to prepare the crude liposomes;

[0086] S8: Subject the crude liposomes to high-pressure homogenization treatment with a homogenization pressure of 400 bar, a temperature of 60 °C, and a circulation times of 5 times, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0087] Example 6

[0088] A composite vitamin derivative nano-liposome is prepared according to the following steps:

[0089] S1: Take 1.5 parts of the active ingredient retinol palmitate, add 6 parts of 1,3-propanediol, place it at room temperature and stir at 300 rpm for 10 min to obtain the mixed solution 1;

[0090] S2: Take 1.5 parts of the active ingredient ascorbyl tetraisopalmitate, add 6 parts of polyethylene glycol, place it at room temperature and stir at 300 rpm for 10 min to obtain the mixed solution 2;

[0091] S3: Take 3 parts of the active ingredient tocopheryl acetate, add 10 parts of glycerol, place it at room temperature and stir at 300 rpm for 10 min to obtain the mixed solution 3;

[0092] S4: Transfer the mixed solutions 1-3 into an oil phase premixing system, heat to 45 °C, and stir at 300 rpm for 15 min to obtain the mixed solution A;

[0093] S5: Add 1 part of egg yolk lecithin and 3 parts of allantoin to the mixed solution A, raise the temperature to 65 °C, and stir at 300 rpm for 20 min to obtain the mixed solution B;

[0094] S6: Prepare 10 parts of cetyl polyoxyethylene ether phosphate sodium and 0.5 part of phenoxyethanol, add them to 57.5 parts of purified water, and stir and disperse in a 70 °C water bath to obtain the mixed solution C;

[0095] S7: Slowly add mixture B to mixture C at a shearing speed of 8000 rpm, a shearing temperature of 50 °C, and a shearing time of 3 min to obtain crude liposomes;

[0096] S8: Subject the crude liposomes to high-pressure homogenization at a homogenization pressure of 500 bar, a temperature of 65 °C, and a circulation number of 2 times, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0097] Example 7

[0098] A composite vitamin derivative nano-liposome is prepared according to the following steps:

[0099] S1: Take 1.2 parts of the active ingredient retinol palmitate and add 5 parts of 1,2-pentanediol; stir at a normal temperature of 300 rpm for 15 min to obtain mixture 1;

[0100] S2: Take 1.2 parts of the active ingredient ascorbyl tetraisopalmitate and add 5 parts of polyethylene glycol, stir at a normal temperature of 100 - 300 rpm for 10 - 30 min to obtain mixture 2;

[0101] S3: Take 3.6 parts of the active ingredient tocopheryl acetate and add 10 parts of glycerol, stir at a normal temperature of 300 rpm for 15 min to obtain mixture 3;

[0102] S4: Transfer mixtures 1 - 3 to an oil-phase premixing system, heat to 45 °C, and stir at 300 rpm for 15 min to obtain mixture A;

[0103] S5: Add 1 part of egg yolk lecithin and 3 parts of allantoin to mixture A, raise the temperature to 65 °C, and stir at 300 rpm for 20 min to obtain mixture B;

[0104] S6: Prepare 10 parts of cetyl polyoxyethylene ether phosphate sodium and 0.5 part of phenoxyethanol, add them to 59.5 parts of purified water, and obtain mixture C in a 70 °C water bath;

[0105] S7: Under shearing conditions, slowly add mixture B to mixture C at a shearing speed of 7000 rpm, a shearing temperature of 50 °C, and a shearing time of 4 min to obtain crude liposomes;

[0106] S8: Subject the crude liposomes to high-pressure homogenization at a homogenization pressure of 600 bar, a temperature of 50 °C, and a circulation number of 2 times, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0107] Comparative Example 1

[0108] Adjust the S2 active substance in Example 1 to "Take 3 parts of the active ingredient retinol palmitate and add 12 parts of 1,2-propanediol; take 3 parts of the active ingredient tocopheryl acetate and add 12 parts of glycerin", and keep other process steps unchanged.

[0109] Comparative Example 2

[0110] Adjust the S2 active substance in Example 1 to "Take 3 parts of the active ingredient retinol palmitate and add 12 parts of 1,2-propanediol; take 3 parts of the active ingredient ascorbyl tetraisopalmitate and add 12 parts of glycerin", and keep other process steps unchanged.

[0111] Comparative Example 3

[0112] Adjust the S2 active substance in Example 1 to "Take 3 parts of the active ingredient ascorbyl tetraisopalmitate and add 12 parts of 1,2-propanediol; take 3 parts of the active ingredient tocopheryl acetate and add 12 parts of glycerin", and keep other process steps unchanged.

[0113] Comparative Example 4

[0114] Adjust the S2 active substance in Example 1 to "Take 6 parts of the active ingredient retinol palmitate and add 24 parts of 1,2-propanediol", and keep other process steps unchanged.

[0115] Comparative Example 5

[0116] Adjust the S2 active substance in Example 1 to "Take 6 parts of the active ingredient ascorbyl tetraisopalmitate and add 24 parts of 1,2-propanediol", and keep other process steps unchanged.

[0117] Comparative Example 6

[0118] Adjust the S2 active substance in Example 1 to "Take 6 parts of the active ingredient tocopheryl acetate and add 24 parts of 1,2-propanediol", and keep other process steps unchanged.

[0119] Comparative Example 7

[0120] A composite vitamin derivative nanoliposome is prepared according to the following steps:

[0121] S1: Take 2 parts of the active ingredient retinol palmitate, 2 parts of the active ingredient ascorbyl tetraisopalmitate, and 2 parts of the active ingredient tocopheryl acetate, add them to 24 parts of 1,2-propanediol, stir at room temperature at 100 rpm for 20 min, then heat to 40 °C and continue to stir at 100 rpm for 10 min to obtain a mixed solution A;

[0122] S2: Add 2 parts of hydrogenated lecithin and 3 parts of hexyl decanol to the mixed solution A, raise the temperature to 60 °C, and stir at 100 rpm for 20 min to obtain a mixed solution B;

[0123] S3: Prepare 5 parts of glyceryl stearate and 0.5 part of potassium sorbate, add them to 59.5 parts of purified water, and stir and disperse in a water bath at 50 °C to obtain mixture C;

[0124] S4: Under shear conditions, slowly add mixture B to mixture C, with a shear speed of 6000 rpm; the shear temperature is 50 °C; the shear time is 5 min to prepare crude liposomes;

[0125] S5: Subject the crude liposomes to high-pressure homogenization treatment, with a homogenization pressure of 500 bar and a temperature of 50 °C; the number of cycles is 2 times, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0126] Comparative Example 8

[0127] A composite vitamin derivative nano-liposome is prepared according to the following steps:

[0128] S1: Take 2 parts of the active ingredient retinol palmitate, 2 parts of the active ingredient ascorbyl tetraisopalmitate, 2 parts of the active ingredient tocopheryl acetate, 16 parts of 1,2-propanediol, 8 parts of glycerol; 2 parts of hydrogenated lecithin and 3 parts of hexyl decanol, mix evenly, stir at room temperature for 10 min and heat to 50 °C, and stir at 100 rpm for 10 min to obtain mixture A;

[0129] S2: Prepare 5 parts of glyceryl stearate and 0.5 part of potassium sorbate, add them to 59.5 parts of purified water, and stir and disperse in a water bath at 50 °C to obtain mixture B;

[0130] S3: Slowly add mixture B to mixture C, stir at room temperature for 60 min; then carry out high-pressure homogenization treatment, with a homogenization pressure of 500 bar and a temperature of 50 °C; the number of cycles is 2 times, and cool at room temperature for 2 h to obtain the composite vitamin derivative liposomes.

[0131] Test Example 1

[0132] Stability test

[0133] Respectively place the samples of Examples 1-7 prepared in the present invention in airtight containers, place them at -18 °C, 4 °C, room temperature and 45 °C for 90 days. At the initial state and after 90 days, check whether there is precipitation or stratification of the sample properties at room temperature, and test the sample retention rate and particle size change, as Figure 1 . It is found by observation that all the samples of the examples show good stability. After being placed at -18 °C, 4 °C, room temperature and 45 °C for 90 days, there is no agglomeration and stratification phenomenon, and there is no leakage of active ingredients. As Figure 2, is a 5% diluted aqueous solution of Example 1, and this aqueous solution is homogeneous and transparent. Therefore, the liposome solution provided by the present invention has good stability and can be stably compounded in the finished product formulation.

[0134] Test Example 2

[0135] Irritation test

[0136] Twenty volunteers aged between 18 and 45 were selected, and it was required that the skin of the volunteers was normal without any special conditions such as drug allergy history. Among them, there were 12 males and 18 females. Examples 1, 3, 5, and 7 were added to the basic emulsion at an addition amount of 5%, and at the same time, the basic emulsion was used as a blank control group for patch testing, and observations were carried out at 1 h, 24 h, and 48 h respectively.

[0137] Local skin reaction scoring grades: 0 point, no reaction; 0.5 point, only faint erythema visible; 1 point, mild erythema; 2 points, moderate erythema; 3 points, severe erythema. Record other local reactions, including: edema, papules, blisters, bullae, pustules, pigmentation, exudation, skin reactions beyond the patch range, etc.

[0138] Calculation of the average cumulative irritation index of the subjects:

[0139] CII = sum of local irritation scores of each sample for each subject / number of observation scores (3 times)

[0140] MCII = sum of CII of all subjects for this drug / number of subjects (20 people)

[0141] The basic emulsion formulation is as follows: 60 parts of purified water; 20 parts of glycerol; 10 parts of cetearyl alcohol; 5 parts of palmitic acid; 5 parts of stearic acid.

[0142] It can be seen from Table 1 below that there is no significant difference in the average cumulative irritation index of the active substances prepared from Examples 1 - 7 into liposomes compared with the blank control group (basic emulsion), and it can be determined that the nano - liposomes provided by the present invention have almost no irritation

[0143] Table 1 Average cumulative irritation index of each group of samples

[0144] Sample group Average stimulation index Example 1 0.10±0.06 Example 2 0.07±0.03 Example 3 0.08±0.08 Example 3 0.09±0.10 Example 4 0.11±0.12 Example 5 0.08±0.09 Example 6 0.09±0.11 Example 7 0.12±0.07 Blank control group 0.09±0.01

[0145] Test Example 3

[0146] Effect of the product of the present invention on melanin synthesis

[0147] Normal human keratinocytes HaCaT were placed in a 96-well plate at a density of 3.5×104 cells / well. After culturing for 1 day, the cells were pre-cultured in Humedia KG2 containing 5% liposomes prepared in Example 1 and Comparative Examples 1-8 for 24 h, and a blank control without samples was set. Then, the cells were irradiated with UVB at a dose of 5 mJ / cm2 in Hank's balanced salt solution (HBSS). At the same time, a non-irradiated negative control group was set. After UVB irradiation, the cells were continued to be cultured in fresh Humedia KG2 for 24 h. An enzyme-linked immunosorbent assay (ELISA) kit was used to quantify IL-1α and PGE2 secreted in the culture medium.

[0148] IL-1α is a trigger that triggers melanocyte proliferation and tyrosinase gene expression through ET-1-related signal transduction. PGE2 also stimulates skin pigmentation by activating melanocytes, including cell growth, upregulation of tyrosinase mRNA, and development of dendrites. In addition, IL-1α and PGE2 are regulated by ROS, and IL-1α increases oxidative stress. The test results are as Figure 3 and Figure 4 shown. The samples prepared in the present invention can effectively reduce the production of melanocyte proliferation factors IL-1α and PGE2 induced by UVB in keratinocytes, that is, the ACE liposomes prepared in the present invention can inhibit UVB-induced skin pigmentation through the ROS scavenging pathway. Moreover, by comparing the prepared examples and comparative examples with the same conditions of the blank group, it can be found that there are extremely significant differences in the inhibitory effects on IL-1α and PGE2 in Example 1 (***, p<0.001), which cannot be achieved by Comparative Examples 1-8, indicating that its efficacy is significantly better than that of Comparative Examples 1-8.

[0149] Test Example 4

[0150] Effect of the product of the present invention on scavenging DPPH free radicals

[0151] Take 100 μL of 1×10 -4 mol / L DPPH ethanol solution, add 100 μL of solutions with different concentrations of Example 1 and Comparative Examples 1-8, mix well, and react in the dark at room temperature for 30 min. Then, measure its absorbance (A i ) at 517 nm with an enzyme-labeled instrument, in parallel 3 times. At the same time, mix 100 μL of DPPH and 100 μL of absolute ethanol and measure its absorbance (A o ) according to the above method, and mix 100 μL of the sample solution and 100 μL of absolute ethanol and measure its absorbance (A j ). The scavenging rate (%) of the sample solution for DPPH free radicals = [1 - (A i - A j ) / A o×100%. The higher the clearance rate, the stronger the reducing ability. The results are as Figure 5 shown. As the addition concentration increases, the efficiency of promoting DPPH radical scavenging increases, and there are also certain differences in the effects of different samples. Among Comparative Examples 1-8, the best effect is in Comparative Example 7. The effect of radical scavenging in Example 1 is significantly better than that in Comparative Example 7, with significant differences at addition amounts of 1% and 5% (*, p < 0.05), and very significant differences at an addition amount of 10% (**, p < 0.01). The above results indicate that the liposomes prepared according to the technical features such as the reasonable compounding of the three active substances and the preparation process provided by the present invention are more conducive to the active substances to exert the DPPH radical scavenging effect.

[0152] Test Example 5

[0153] Effect of the product of the present invention on ABTS radical scavenging

[0154] Prepare a 7 mmol / L stock solution of ABTS with distilled water. Take 5 mL and add 88 μL of a 140 mmol / L potassium persulfate stock solution and mix. Place it in the dark at room temperature for 12 h to form an ABTS radical stock solution. Dilute this stock solution with absolute ethanol to a working solution, and adjust the absorbance value to 0.7 ± 0.02 at 734 nm. Take 150 μL of the ABTS radical working solution, add 150 μL of sample solutions of Example 1 and Comparative Examples 1-8 at different concentrations, mix well, and let it stand in the dark at room temperature for 6 min. Then measure the absorbance (A) with a microplate reader at a wavelength of 734 mm, in parallel 3 times. At the same time, mix 150 μL of the ABTS radical working solution and 150 μL of absolute ethanol and measure its absorbance (A0) according to the above method. The clearance rate (%) of the sample solution for ABTS radicals = (1 - A / A0) × 100%. The higher the clearance rate, the stronger the reducing ability. The results are as Figure 6 shown. As the concentration increases, the efficiency of promoting ABTS radical scavenging increases, and there are also certain differences in the effects of different samples. Among Comparative Examples 1-8, the best effect is in Comparative Example 7. The effect of radical scavenging in Example 1 is significantly better than that in Comparative Example 7, with very significant differences at an addition amount of 1% (**, p < 0.01), and very significant differences at addition amounts of 5% and 10% (***, p < 0.001). This result shows that the liposomes prepared by the present invention have excellent ABTS radical scavenging effects.

[0155] Test Example 6

[0156] Effect of the product of the present invention on collagen synthesis

[0157] 2 ml of human skin fibroblasts cultured to the logarithmic phase were implanted into a culture plate with a cover glass at a concentration of 3×105 cells / ml. The cover glass was treated with polylysine before culture. Using DMEM culture medium containing 10% FBS, after culturing for 24 h, 5% diluted aqueous solutions of Example 1 and Comparative Examples 1-8 were prepared for testing. Three parallel samples were set for each concentration. The control group was added with an equal volume of serum, cultured for 7 days, then the cover glass was washed with PBS buffer, fixed with 5% acetone, peroxidase was removed using H2O2, the primary antibody was added, incubated at 37 °C for 2 h, the secondary antibody was added, and diaminobenzidine (DAB) was used for color development. Observation was carried out using an optical microscope. Brownish-yellow was positive. 10 fields of view on the cover glass were randomly selected for observation and analyzed by LEICAQ software to observe the expression of type I collagen. The lower the gray value, the higher the content of type I collagen. The results are as Figure 7 shown. Example 1 had the highest promotion rate of collagen synthesis, and the difference was extremely significant compared with the control group (***, p<0.0001). Comparative Examples 1-8 all had a positive effect on collagen synthesis. Compared with the control group, Comparative Examples 1, 2, 7, and 8 had a relatively significant difference (**, p<0.001), and Comparative Example 3 had a significant difference (*, p<0.05). The remaining samples were considered to have basically no difference. This shows that Example 1 with a combination of 3 active substances is superior to Comparative Examples with only 1 or 2 active substances, and the liposomes of the three active substances under different preparation processes also have a less significant effect on collagen synthesis than Example 1. On the one hand, this result shows the excellent anti-aging effect of the samples prepared by the present invention, and on the other hand, it also shows the synergistic effect between the three active ingredients.

[0158] Test Example 7

[0159] Anti-aging test of human skin

[0160] Test subjects: 30 test subjects (30 - 50 years old) with obvious fine lines or skin relaxation on the face, including 21 females and 9 males. During the test, the two sides of the face were used as a control, and a double-blind test was conducted.

[0161] Test samples: The composite vitamin derivative emulsion sample prepared with 5% of Example 1 was used for testing, with the basic emulsion as a control. The dosage was 2 mg / cm 2 . The formula of the basic emulsion is as follows: 60 parts of purified water; 20 parts of glycerol; 10 parts of cetostearyl alcohol; 5 parts of palmitic acid; 5 parts of stearic acid.

[0162] Evaluation period: The skin condition was tested on the 14th and 28th days after use respectively.

[0163] Evaluation dimensions: The skin elasticity of the subjects and the glossiness of the tested skin.

[0164] Evaluation method: Use Cutometer MPA580 to test skin elasticity and firmness. The larger the ΔQ3, the more obvious the improvement in skin firmness. Use Glossymeter GL200 to test skin gloss. The larger the ΔDSC and ΔGloss values, the better the skin gloss.

[0165] Evaluation results: After continuous use for 28 days, compared with the control group, it was significantly observed that the wrinkles of the subjects decreased (**, indicating p < 0.01, the difference was very significant), and the skin became firmer and more elastic (**, indicating p < 0.01, the difference was very significant); the skin gloss was significantly improved (**, indicating p < 0.01, the difference was very significant). The results are as Figure 8 shown.

[0166] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A composite vitamin derivative nano-liposome, characterized in that, For every 100 parts by weight of the raw materials of the nano-liposomes, the following raw materials are included: 15 - 30 parts of polyol, 3 - 15 parts of compound vitamin derivative, 1 - 3 parts of lecithin, 0.5 - 5 parts of emollient, 5 - 10 parts of emulsifier, 0.5 - 2 parts of preservative, and the balance is purified water.

2. The nano-liposome of a composite vitamin derivative according to claim 1, characterized in that, For every 100 parts by weight of the raw materials of the nano-liposomes, the following raw materials are included: 20 - 30 parts of polyol, 6 - 15 parts of compound vitamin derivative, 1 - 2 parts of lecithin, 0.5 - 3 parts of emollient, 5 - 8 parts of emulsifier, 0.5 - 1 part of preservative, and the balance is purified water.

3. A composite vitamin derivative nano-liposome according to any one of claims 1-2, characterized in that, The compound vitamin derivative is composed of retinyl palmitate, ascorbyl tetraisopalmitate, and tocopheryl acetate with a weight ratio of 1 - 5:1 - 5:1 - 5.

4. A composite vitamin derivative nanoliposome according to any one of claims 1-2, characterized in that, The compound vitamin derivative is composed of retinyl palmitate, ascorbyl tetraisopalmitate, and tocopheryl acetate with a weight ratio of 1:1:

1.

5. A composite vitamin derivative nano-liposome according to any one of claims 1-2, characterized in that, The polyol is one or more of 1,2 - propanediol, 1,3 - propanediol, 1,4 - butanediol, 1,2 - pentanediol, glycerol, and polyethylene glycol.

6. A composite vitamin derivative nano-liposome according to any one of claims 1-2, characterized in that, The emulsifier is one or more of polysorbate - 20, glyceryl stearate, sodium lauryl polyoxyethylene ether sulfate, sodium hexadecyl polyoxyethylene ether phosphate, C12 - 14 alkyl dimethyl benzyl ammonium chloride, cocamidopropyl betaine, and sorbitan monolaurate.

7. A composite vitamin derivative nano-liposome according to any one of claims 1-2, characterized in that, The emollient is one or more of cholesterol, allantoin, hexyl decanol, octyldecanol, cetyl alcohol, and isopropyl palmitate.

8. A composite vitamin derivative nano-liposome according to any one of claims 1-2, characterized in that, The lecithin is one or more of hydrogenated lecithin, soybean lecithin, and egg yolk lecithin.

9. A composite vitamin derivative nanoliposome according to any one of claims 1-2, characterized in that, The preservative is one or more of phenoxyethanol, potassium sorbate, glycerol caprate, capric acid, glycerol caprylate, and sorbitan octanoate.

10. A method for preparing a composite vitamin derivative nano-liposome, characterized in that, The nano-liposomes are prepared according to the following steps: S1: Take 1 - 5 parts of the active ingredient retinyl palmitate and add 5 - 10 parts of polyol; stir at a normal temperature of 100 - 300 rpm for 10 - 30 min to obtain mixture 1. S2: Take 1 - 5 parts of the active ingredient ascorbyl tetraisopalmitate and add 5 - 10 parts of polyol; stir at a normal temperature of 100 - 300 rpm for 10 - 30 min to obtain mixture 2. S3: Take 1 - 5 parts of the active ingredient tocopheryl acetate and add 5 - 10 parts of polyol; stir at a normal temperature of 100 - 300 rpm for 10 - 30 min to obtain mixture 3. S4: Transfer mixtures 1 - 3 to an oil phase premixing system, heat to 30 - 50 °C, and stir at 100 - 500 rpm for 10 - 30 min to obtain mixture A. S5: Add 1 - 3 parts of lecithin and 0.5 - 5 parts of emollient to mixture A, raise the temperature to 60 - 65 °C, and continue to stir at 100 - 500 rpm for 10 - 30 min to obtain mixture B. S6: Prepare 5 - 10 parts of emulsifier and 0.5 - 2 parts of preservative, add them to pure water, and stir and disperse in a water bath at 50 - 70 °C to obtain mixture C. S7: Slowly add mixture B to mixture C, and set the shear rotation speed to 5000 - 8000 rpm; the shear temperature to 50 - 70 °C; and the shear time to 3 - 8 min to obtain crude liposomes. S8: Subject the crude liposomes to high-pressure homogenization treatment, with a homogenization pressure of 400 - 800 bar and a temperature of 50 - 65 °C; the number of cycles is 1 - 5 times, and cool at room temperature for at least 2 h to obtain the composite vitamin derivative liposomes.

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