An anti-aging enhanced PDRN composite composition and its preparation method

Abstract

This invention belongs to the field of nucleotide composition technology and relates to an anti-aging enhanced PDRN composite composition and its preparation method, including the following steps: preparation of a highly stable PDRN complex; preparation of an anti-aging enhanced liquid crystal oil phase carrier; and assembly of a PDRN aqueous solution and a liquid crystal oil phase solution. In this invention, glyceryl monooleate, cetyl-PG hydroxyethyl palmitamide, and behenol are mixed and heated. After cooling, capryloyl salicylic acid and β-glucan-pterostilbene solution are added and mixed evenly. The mixture is then cooled at a constant rate to initially form a layered liquid crystal oil phase in the solution. This is followed by rapid cooling to stabilize the liquid crystal structure. The resulting liquid crystal oil phase enables the subsequent water-in-oil system to have high stability. Cetyl-PG hydroxyethyl palmitamide and capryloyl salicylic acid synergistically increase the transdermal permeability of the water-in-oil system, while pterostilbene and β-glucan enhance the sustained-release stability of the system, thus synergistically improving the anti-aging ability of the PDRN composite composition on the skin.

Description

Technical Field

[0001] This invention belongs to the field of nucleotide composition technology, specifically relating to an anti-aging enhanced PDRN composite composition and its preparation method. Background Technology

[0002] Polydeoxyribonucleotides (PDRNs) are specific DNA fragments extracted from salmon germ cells or salmon seminal vesicles. They have a 98% base similarity to human DNA and can generate nucleotides and nucleosides that accelerate DNA synthesis, enhance fibroblast proliferation and growth, promote skin tissue regeneration, inhibit the expression of inflammatory cytokines and apoptotic proteins, and have anti-inflammatory and UV-damage-resistant effects. However, if the molecular weight of PDRN is too small, it is difficult to bind effectively to A2A receptors, thus reducing its anti-aging effects. If the molecular weight of PDRN is too large and highly polar, it is difficult to penetrate the skin barrier and accumulate at the anti-aging target sites in the skin, thus also reducing its anti-aging effects.

[0003] To fully utilize the efficacy of PDRN, current technologies typically employ microneedle injection to inject it into subcutaneous tissue or prepare it as nanoliposomes for penetration. However, microneedle injection causes some damage to the skin, potentially leading to discomfort after injection. Furthermore, it requires specialized personnel and has limitations. As for preparing PDRN into nanoliposomes, current technologies often use conventional oil-phase systems to directly encapsulate aqueous PDRN. The resulting water-in-oil system has poor stability, and the activity of PDRN is easily lost during storage. Summary of the Invention

[0004] To address the aforementioned technical deficiencies, this invention presents an anti-aging enhanced PDRN composite composition and its preparation method. First, PDRN is composite modified and an anti-aging enhanced liquid crystal oil phase carrier is prepared. Then, an oil-in-water system is constructed, which can significantly enhance the stability and sustained-release performance of the system, fully leveraging the anti-aging and skin-repairing effects of the system.

[0005] A method for preparing an anti-aging enhanced PDRN composite composition includes the following steps:

[0006] S1: Preparation of highly stable PDRN complex

[0007] By weight, 1-1.2 parts of PDRN and hydroxyethylpiperazine ethane sulfonate buffer were combined to prepare PDRN stock solution. Then, 0.8-1 parts of silk core protein were added, followed by 0.08-0.1 parts of trimethylpentanediol / adipic acid / glycerol crosspolymer and 0.2-0.25 parts of PPG-24-glycerol polyether-24. The mixture was then sonicated, filtered, and cleaned to obtain a highly stable PDRN complex.

[0008] S2: Preparation of Anti-aging Enhanced Liquid Crystal Oil Phase Support

[0009] By weight, 1-1.2 parts of β-glucan, 0.2-0.3 parts of lecithin and 0.3-0.5 parts of pterostilbene are mixed evenly and filtered to obtain a β-glucan-pterostilbene solution. 10-12 parts of glyceryl monooleate, 3.5-4 parts of cetyl-PG hydroxyethyl palmitamide and 2-3 parts of behenol are heated, mixed evenly and then cooled. 0.8-1 parts of capryloyl salicylic acid and the β-glucan-pterostilbene solution are added, and the mixture is cooled rapidly after uniform cooling to obtain a liquid crystal oil phase solution.

[0010] S3: Assembly of PDRN aqueous solution and liquid crystal oil solution

[0011] By weight, 0.08-0.1 parts of a highly stable PDRN complex are mixed with 10-15 parts of purified water to prepare an aqueous solution, which is then slowly added dropwise to 50-60 parts of a liquid crystal oil solution to obtain an anti-aging enhanced PDRN composite composition.

[0012] Furthermore, the preparation of the highly stable PDRN complex in step S1 specifically includes the following steps:

[0013] S1.1: Place 1-1.2 parts of PDRN in a glass reactor, add 10-12 parts of hydroxyethylpiperazine ethane sulfonic acid buffer solution with pH 7.4, and stir at 200-250 rpm for 20-25 minutes to obtain PDRN mother liquor;

[0014] S1.2: Under high-speed stirring conditions of 5-10℃ and 4000-5000rpm, 0.8-1 part of fibroin was added to the PDRN mother liquor in step S1.1 in 3-4 portions to obtain the precursor solution.

[0015] S1.3: Take the precursor solution obtained in step S1.2, add 0.08-0.1 parts of trimethylpentanediol / adipic acid / glycerol cross-linking polymer and 0.2-0.25 parts of PPG-24-glycerol polyether-24, transfer to an ultrasonic cell disruptor, sonicate at a frequency of 35-40 kHz for 12-15 minutes under a nitrogen atmosphere, let stand for 20-30 minutes, filter, take the solid and wash it with deionized water to obtain a highly stable PDRN complex.

[0016] Furthermore, the preparation of the anti-aging enhanced liquid crystal oil phase carrier in step S2 specifically includes the following steps:

[0017] S2.1: Dissolve 1-1.2 parts of β-glucan in 15-20 parts of deionized water at 60-65℃, add 0.2-0.3 parts of lecithin, sonicate at a frequency of 25-30kHz for 15-20 minutes, then add 0.3-0.5 parts of pterostilbene, and magnetically stir at 60-65℃ and 500-600rpm for 30-35 minutes. Filter through a microporous membrane to obtain a β-glucan-pterostilbene solution.

[0018] S2.2: Add 10-12 parts of glyceryl monooleate, 3.5-4 parts of cetyl-PG hydroxyethyl palmitamide and 2-3 parts of behenol to a vacuum emulsification reactor. Stir at 70-75℃ and 200-250 rpm for 25-30 minutes. Then cool to 40-45℃, add 0.8-1 parts of capryloyl salicylic acid and the β-glucan-pterostilbene solution prepared in step S2.1, and stir at a constant temperature of 150-200 rpm for 30-40 minutes. After uniformly cooling to 30-35℃, inject cold nitrogen gas at 5℃ to rapidly cool to 8-10℃. Hold at this temperature for 10-15 minutes to obtain a liquid crystal oil phase solution.

[0019] Furthermore, the assembly of the S3PDRN aqueous solution and the liquid crystal oil solution specifically includes the following steps:

[0020] S3.1: Mix 0.08-0.1 parts of the highly stable PDRN complex with 10-15 parts of purified water until homogeneous to obtain an aqueous solution;

[0021] S3.2: Take 50-60 parts of liquid crystal oil phase solution and heat it to 37-40℃. Then, add the aqueous phase solution obtained in step S3.1 dropwise to the liquid crystal oil phase solution within 30-40 minutes to obtain a nanoemulsion, which is the anti-aging enhanced PDRN composite composition.

[0022] Furthermore, the PDRN in step S1.1 has a purity ≥96% and an average molecular weight of 200-850 kDa.

[0023] Furthermore, the temperature of the hydroxyethylpiperazine ethane sulfonic acid buffer solution in step S1.1 is 3-4°C.

[0024] Furthermore, in step S1.2, the time interval between each addition of the silk protein is 4-5 minutes.

[0025] Furthermore, the pore size of the microporous filter membrane in step S2.1 is 0.45-0.5 μm.

[0026] Furthermore, the cooling rate in step S2.2 is 1-1.5℃ / min.

[0027] An anti-aging enhanced PDRN composite composition is prepared by the above-described method for preparing an anti-aging enhanced PDRN composite composition.

[0028] The beneficial effects of this invention are:

[0029] 1. This invention first modifies β-glucan with lecithin, then adds pterostilbene and stirs to obtain a β-glucan-pterostilbene solution. Then, glyceryl monooleate, cetyl-PG hydroxyethyl palmitamide, and behenol are mixed and heated. After cooling, capryloyl salicylic acid and the β-glucan-pterostilbene solution are added and mixed evenly. Then, the mixture is cooled at a constant rate to initially form a layered liquid crystal oil phase in the solution. Then, rapid cooling is performed to fix the liquid crystal structure. The obtained liquid crystal oil phase can give the subsequent water-in-oil system high stability. Cetyl-PG hydroxyethyl palmitamide and capryloyl salicylic acid can synergistically increase the transdermal rate of the water-in-oil system. Pterostilbene and β-glucan can enhance the sustained-release stability of the system. Cetyl-PG hydroxyethyl palmitamide, capryloyl salicylic acid, pterostilbene, and β-glucan can also synergistically improve the anti-aging ability of the PDRN composite composition on the skin.

[0030] 2. This invention adds trimethylpentanediol / adipic acid / glycerol cross-linking polymer and PPG-24-glycerol polyether-24 to a mixture of polydeoxyribonucleotides and fibroin. Under the cross-linking network formed by PPG-24-glycerol polyether-24 and trimethylpentanediol / adipic acid / glycerol cross-linking polymer, polydeoxyribonucleotides and fibroin can form a highly stable complex, thereby enhancing the storage stability of PDRN in the subsequent water-in-oil system, ensuring the activity of PDRN when it enters the dermis, and improving the skin repair effect of PDRN. Attached Figure Description

[0031] Figure 1 This is a flowchart illustrating the preparation method of the anti-aging enhanced PDRN composite composition used in the embodiments of the present invention. Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] It should be noted that, unless otherwise specified, all parts in the following embodiments represent parts by weight.

[0034] Example 1

[0035] A method for preparing an anti-aging enhanced PDRN composite composition, such as... Figure 1 As shown, the specific steps include:

[0036] S1: Preparation of highly stable PDRN complex

[0037] S1.1: Place 1 part of PDRN in a glass reactor. The purity of PDRN is 96% and the average molecular weight is 200 kDa. Add 10 parts of hydroxyethylpiperazine ethane sulfonic acid buffer with pH 7.4. The temperature of the hydroxyethylpiperazine ethane sulfonic acid buffer is 3°C. Stir at 200 rpm for 20 minutes to obtain PDRN mother liquor.

[0038] S1.2: Under high-speed stirring conditions of 5℃ and 4000rpm, 0.8 parts of fibroin were added to the PDRN mother liquor in step S1.1 in 3 portions, with an interval of 4 minutes between each addition of fibroin, to obtain the precursor solution.

[0039] S1.3: Take the precursor solution obtained in step S1.2, add 0.08 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer and 0.2 parts of PPG-24-glycerol polyether-24, transfer to an ultrasonic cell disruptor, sonicate at a frequency of 35 kHz for 12 minutes under a nitrogen atmosphere, let stand for 20 minutes and then filter, take the solid and wash it with deionized water to obtain a highly stable PDRN complex.

[0040] S2: Preparation of Anti-aging Enhanced Liquid Crystal Oil Phase Support

[0041] S2.1: Dissolve 1 part of β-glucan in 15 parts of deionized water at 60℃, add 0.2 parts of lecithin, sonicate at 25kHz for 15 minutes, then add 0.3 parts of pterostilbene, and magnetically stir at 60℃ and 500rpm for 30 minutes. Filter through a microporous membrane with a pore size of 0.45μm to obtain a β-glucan-pterostilbene solution.

[0042] S2.2: Add 10 parts of glyceryl monooleate, 3.5 parts of cetyl-PG hydroxyethyl palmitamide and 2 parts of behenol to a vacuum emulsification reactor. Stir at 70°C and 200 rpm for 25 minutes, then cool to 40°C. Add 0.8 parts of capryloyl salicylic acid and the β-glucan-pterostilbene solution prepared in step S2.1. Stir at a constant temperature of 150 rpm for 30 minutes. Cool to 30°C at a uniform cooling rate of 1°C / min, then inject cold nitrogen gas at 5°C to rapidly cool to 8°C. Hold for 10 minutes to obtain a liquid crystal oil phase solution.

[0043] S3: Assembly of PDRN aqueous solution and liquid crystal oil solution

[0044] S3.1: Mix 0.08 parts of the highly stable PDRN complex with 10 parts of purified water until homogeneous to obtain an aqueous solution;

[0045] S3.2: Take 50 parts of liquid crystal oil phase solution and heat it to 37°C. Then, add the aqueous phase solution obtained in step S3.1 dropwise to the liquid crystal oil phase solution within 30 minutes to obtain a nanoemulsion, which is the anti-aging enhanced PDRN composite composition.

[0046] Example 2

[0047] A method for preparing an anti-aging enhanced PDRN composite composition, such as... Figure 1 As shown, the specific steps include:

[0048] S1: Preparation of highly stable PDRN complex

[0049] S1.1: Place 1.2 parts of PDRN in a glass reactor. The purity of PDRN is 96% and the average molecular weight is 200 kDa. Add 12 parts of hydroxyethylpiperazine ethane sulfonic acid buffer with a pH of 7.4. The temperature of the hydroxyethylpiperazine ethane sulfonic acid buffer is 3°C. Stir at 200 rpm for 20 minutes to obtain PDRN mother liquor.

[0050] S1.2: Under high-speed stirring conditions of 5℃ and 4000rpm, 1 part of silk protein was added to the PDRN mother liquor in step S1.1 in 3 portions, with an interval of 4 minutes between each addition of silk protein, to obtain the precursor solution.

[0051] S1.3: Take the precursor solution obtained in step S1.2, add 0.1 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer and 0.25 parts of PPG-24-glycerol polyether-24, transfer to an ultrasonic cell disruptor, sonicate at a frequency of 35 kHz for 12 minutes under a nitrogen atmosphere, let stand for 20 minutes and then filter, take the solid and wash it with deionized water to obtain a highly stable PDRN complex.

[0052] S2: Preparation of Anti-aging Enhanced Liquid Crystal Oil Phase Support

[0053] S2.1: Dissolve 1.2 parts of β-glucan in 20 parts of deionized water at 60℃, add 0.3 parts of lecithin, sonicate at 25kHz for 15 minutes, then add 0.5 parts of pterostilbene, and magnetically stir at 60℃ and 500rpm for 30 minutes. Filter through a microporous membrane with a pore size of 0.45μm to obtain a β-glucan-pterostilbene solution.

[0054] S2.2: Add 12 parts of glyceryl monooleate, 4 parts of cetyl-PG hydroxyethyl palmitamide and 3 parts of behenol to a vacuum emulsification reactor. Stir at 70°C and 200 rpm for 25 minutes, then cool to 40°C. Add 1 part of capryloyl salicylic acid and the β-glucan-pterostilbene solution prepared in step S2.1. Stir at a constant temperature of 150 rpm for 30 minutes. Cool to 30°C at a uniform cooling rate of 1°C / min, then inject cold nitrogen gas at 5°C to rapidly cool to 8°C. Hold for 10 minutes to obtain a liquid crystal oil phase solution.

[0055] S3: Assembly of PDRN aqueous solution and liquid crystal oil solution

[0056] S3.1: Mix 0.1 parts of the highly stable PDRN complex with 15 parts of purified water until homogeneous to obtain an aqueous solution;

[0057] S3.2: Take 60 parts of liquid crystal oil phase solution and heat it to 37°C. Then, add the aqueous phase solution obtained in step S3.1 dropwise to the liquid crystal oil phase solution within 30 minutes to obtain a nanoemulsion, which is the anti-aging enhanced PDRN composite composition.

[0058] Example 3

[0059] A method for preparing an anti-aging enhanced PDRN composite composition, such as... Figure 1 As shown, the specific steps include:

[0060] S1: Preparation of highly stable PDRN complex

[0061] S1.1: Place 1 part of PDRN in a glass reactor. The purity of PDRN is 99% and the average molecular weight is 850 kDa. Add 10 parts of hydroxyethylpiperazine ethane sulfonic acid buffer with pH 7.4. The temperature of the hydroxyethylpiperazine ethane sulfonic acid buffer is 4°C. Stir at 250 rpm for 25 minutes to obtain PDRN mother liquor.

[0062] S1.2: Under high-speed stirring conditions of 10℃ and 5000rpm, 0.8 parts of fibroin were added to the PDRN mother liquor in step S1.1 in 4 portions, with an interval of 5 minutes between each addition of fibroin, to obtain the precursor solution.

[0063] S1.3: Take the precursor solution obtained in step S1.2, add 0.08 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer and 0.2 parts of PPG-24-glycerol polyether-24, transfer to an ultrasonic cell disruptor, sonicate at a frequency of 40 kHz for 15 minutes under a nitrogen atmosphere, let stand for 30 minutes and then filter, take the solid and wash it with deionized water to obtain a highly stable PDRN complex.

[0064] S2: Preparation of Anti-aging Enhanced Liquid Crystal Oil Phase Support

[0065] S2.1: Dissolve 1 part of β-glucan in 15 parts of deionized water at 65℃, add 0.2 parts of lecithin, sonicate at 30kHz for 15 minutes, then add 0.3 parts of pterostilbene, and magnetically stir at 65℃ and 600rpm for 35 minutes. Filter through a microporous membrane with a pore size of 0.5μm to obtain a β-glucan-pterostilbene solution.

[0066] S2.2: Add 10 parts of glyceryl monooleate, 3.5 parts of cetyl-PG hydroxyethyl palmitamide and 2 parts of behenol to a vacuum emulsification reactor. Stir at 75°C and 250 rpm for 30 minutes, then cool to 45°C. Add 0.8 parts of capryloyl salicylic acid and the β-glucan-pterostilbene solution prepared in step S2.1. Stir at a constant temperature of 200 rpm for 40 minutes. Cool to 35°C at a cooling rate of 1.5°C / min, then inject cold nitrogen gas at 5°C to rapidly cool to 10°C. Hold at this temperature for 15 minutes to obtain a liquid crystal oil phase solution.

[0067] S3: Assembly of PDRN aqueous solution and liquid crystal oil solution

[0068] S3.1: Mix 0.08 parts of the highly stable PDRN complex with 10 parts of purified water until homogeneous to obtain an aqueous solution;

[0069] S3.2: Take 50 parts of liquid crystal oil phase solution and heat it to 40°C. Then, add the aqueous phase solution obtained in step S3.1 dropwise to the liquid crystal oil phase solution within 40 minutes to obtain a nanoemulsion, which is the anti-aging enhanced PDRN composite composition.

[0070] Comparative Example 1

[0071] The difference from Example 1 is that Comparative Example 1 uses a conventional water-in-oil system, and the liquid crystal oil phase solution in S3 is replaced with a conventional oil phase solution (Span 80, isopropyl palmitate and Tween 80 are weighed in a mass ratio of 1:1:1.5, Span 80 and isopropyl palmitate are heated and mixed at 80°C, and after cooling to 37°C, Tween 80 is added and stirred and mixed). The rest of the specific implementation methods are the same as those in Example 1.

[0072] Comparative Example 2

[0073] The difference from Example 1 is that Comparative Example 2 omits step S1.2, replaces the precursor solution in step S1.3 with PDRN mother liquor to obtain the PDRN complex, and replaces the high-stability PDRN complex in step S3 with an equal mass of the PDRN complex. All other specific implementation methods are the same as in Example 1.

[0074] Experiment 1: The anti-aging enhanced PDRN composite compositions prepared in Examples 1-3 and the PDRN composite compositions prepared in Comparative Examples 1-2 were used as samples and stored at room temperature. The Zeta potential value and average particle size of the samples were tested after storage times of 1, 4 and 8 months, respectively. Three parallel experiments were conducted and the average value was taken. The results are shown in Table 1.

[0075] Table 1: Zeta potential and average particle size after sample storage for different times

[0076]

[0077] It is known that the higher the Zeta potential value, the better the system stability and the smaller the increase in particle size during storage. As can be seen from the data of Examples 1-3 and Comparative Examples 1-2 in Table 1, the anti-aging enhanced PDRN composite compositions prepared in Examples 1-3 have high stability. Meanwhile, the stability of Comparative Examples 1-2 is lower than that of Examples 1-3. This proves that preparing a liquid crystal oil phase solution as the oil phase for water-in-oil can improve the stability of the obtained anti-aging enhanced PDRN composite compositions. Moreover, using fibroin to modify PDRN can also improve the stability of the anti-aging enhanced PDRN composite compositions.

[0078] Experiment 2: The anti-aging enhanced PDRN composite compositions prepared in Examples 1-3 and the PDRN composite compositions prepared in Comparative Examples 1-2 were mixed with physiological saline at a mass ratio of 1:10 to obtain test samples. Hair was removed from the same area on the back of 10 SD rats. The skin moisture content (skin moisture meter), skin elasticity (skin elasticity meter), and wrinkle depth (skin surface 3D scanner) of the hair-removed area were measured and the average value was recorded as the initial value. Then, the test sample was applied twice a day, 2 mL each time, for 30 consecutive days. The skin moisture content, skin elasticity, and wrinkle depth were measured again and the average value was recorded as the final value. The change value = final value - initial value was recorded. The results are shown in Table 2.

[0079] Table 2: Anti-aging effects of test samples on skin

[0080]

[0081] As can be seen from the data in Table 2 of Examples 1-3 and Comparative Examples 1-2, the anti-aging enhanced PDRN composite compositions prepared in Examples 1-3 have excellent anti-aging effects on the skin. However, the anti-aging effect of Comparative Examples 1-2 is significantly reduced compared to Examples 1-3. This demonstrates that preparing a liquid crystal oil phase solution as the oil phase for water-in-oil emulsion can enhance the anti-aging effect of the prepared PDRN composite compositions. Furthermore, using fibroin to modify PDRN can also enhance the anti-aging effect of the PDRN composite compositions.

[0082] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A method for preparing an anti-aging enhanced PDRN composite composition, characterized in that, Includes the following steps: S1: Preparation of highly stable PDRN complex S1.1: Place 1-1.2 parts of PDRN in a glass reactor, add 10-12 parts of hydroxyethylpiperazine ethane sulfonic acid buffer solution with pH 7.4, and stir at 200-250 rpm for 20-25 minutes to obtain PDRN mother liquor; S1.2: Under high-speed stirring conditions of 5-10℃ and 4000-5000rpm, 0.8-1 part of fibroin was added to the PDRN mother liquor in step S1.1 in 3-4 portions to obtain the precursor solution. S1.3: Take the precursor solution obtained in step S1.2, add 0.08-0.1 parts of trimethylpentanediol / adipic acid / glycerol cross-linking polymer and 0.2-0.25 parts of PPG-24-glycerol polyether-24, transfer to an ultrasonic cell disruptor, sonicate at a frequency of 35-40 kHz for 12-15 minutes under a nitrogen atmosphere, let stand for 20-30 minutes, filter, take the solid and wash it with deionized water to obtain a highly stable PDRN complex; S2: Preparation of Anti-aging Enhanced Liquid Crystal Oil Phase Support S2.1: Dissolve 1-1.2 parts of β-glucan in 15-20 parts of deionized water at 60-65℃, add 0.2-0.3 parts of lecithin, sonicate at a frequency of 25-30kHz for 15-20 minutes, then add 0.3-0.5 parts of pterostilbene, and magnetically stir at 60-65℃ and 500-600rpm for 30-35 minutes. Filter through a microporous membrane to obtain a β-glucan-pterostilbene solution. S2.2: Add 10-12 parts of glyceryl monooleate, 3.5-4 parts of cetyl-PG hydroxyethyl palmitamide and 2-3 parts of behenol to a vacuum emulsification reactor. Stir at 70-75℃ and 200-250rpm for 25-30 minutes. Then cool to 40-45℃, add 0.8-1 parts of capryloyl salicylic acid and the β-glucan-pterostilbene solution prepared in step S2.

1. Stir at a constant temperature of 150-200rpm for 30-40 minutes. After cooling to 30-35℃, inject cold nitrogen gas at 5℃ to rapidly cool to 8-10℃. Hold at this temperature for 10-15 minutes to obtain a liquid crystal oil phase solution. S3: Assembly of PDRN aqueous solution and liquid crystal oil solution By weight, 0.08-0.1 parts of a highly stable PDRN complex are mixed with 10-15 parts of purified water to prepare an aqueous solution, which is then slowly added dropwise to 50-60 parts of a liquid crystal oil solution to obtain an anti-aging enhanced PDRN composite composition.

2. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, The assembly of the S3PDRN aqueous solution and the liquid crystal oil solution specifically includes the following steps: S3.1: Mix 0.08-0.1 parts of the highly stable PDRN complex with 10-15 parts of purified water until homogeneous to obtain an aqueous solution; S3.2: Take 50-60 parts of liquid crystal oil phase solution and heat it to 37-40℃. Then, add the aqueous phase solution obtained in step S3.1 dropwise to the liquid crystal oil phase solution within 30-40 minutes to obtain a nanoemulsion, which is the anti-aging enhanced PDRN composite composition.

3. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, The PDRN in step S1.1 has a purity of ≥96% and an average molecular weight of 200-850 kDa.

4. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, The temperature of the hydroxyethylpiperazine ethane sulfonic acid buffer solution in step S1.1 is 3-4℃.

5. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, In step S1.2, the time interval between each addition of silk protein is 4-5 minutes.

6. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, The pore size of the microporous filter membrane in step S2.1 is 0.45-0.5 μm.

7. The method for preparing an anti-aging enhanced PDRN composite composition according to claim 1, characterized in that, The cooling rate in step S2.2 is 1-1.5℃ / min.

8. A PDRN composite composition with enhanced anti-aging properties, characterized in that, It is prepared by the method of any one of claims 1-7 for preparing an anti-aging enhanced PDRN composite composition.

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