A penetration enhancing composition, polydeoxyribonucleotide essence and method of preparation thereof

Abstract

The application discloses a penetration-promoting composition, a polydeoxyribonucleotide essence and a preparation method thereof, relates to the cosmetic technical field, and the preparation method of the penetration-promoting composition comprises the following steps: preparing a hydroxyethyl deacetylated chitosan solution and a polydeoxyribonucleotide solution; adding the hydroxyethyl deacetylated chitosan solution into the polydeoxyribonucleotide solution, stirring, then adding isosorbide dimethyl ether, and stirring again until uniform. The penetration-promoting effect is improved by the synergistic effect of the aggregate of the hydroxyethyl deacetylated chitosan and the polydeoxyribonucleotide and the isosorbide dimethyl ether, a great breakthrough is realized in the relative penetration amount and the penetration depth, the polydeoxyribonucleotide can reach the DEJ layer, which is a key effective target point position of the polydeoxyribonucleotide in the skin, adjusts the basal cells, and then reaches the dermis layer to activate fibroblasts, so that the bioavailability and the repair and anti-aging effect of the polydeoxyribonucleotide in the skin are greatly improved.

Description

Technical Field

[0001] This invention relates to a penetration-enhancing composition, a polydeoxyribonucleic acid extract, and a method for preparing the same, belonging to the field of cosmetic technology. Background Technology

[0002] Polydeoxyribonucleotide (PDRN) is a naturally derived DNA derivative with a molecular weight ranging from 50 to 1500 kDa. It has no antigenic properties and no systemic toxicity. Currently, sources for extraction and purification include human placenta, rose, red algae, ginseng, and sperm cells from salmon (golden trout) or salmon (salmon). Due to economic efficiency and the complexity of the extraction process, PDRN extracted from fish reproductive cells has become the primary source.

[0003] Polydeoxyribonucleotides (PDRNs) are recognized as A2A receptor agonists. A2A receptors are crucial drivers of skin metabolism. Studies of real human skin tissue have revealed that A2A receptors are not uniformly distributed throughout the skin, but rather exhibit spatial specificity. A2A receptors are primarily expressed at the base of basal keratinocytes, especially on the side facing the basement membrane. Furthermore, A2A receptors have also been observed in vascular endothelial cells of the dermis. Activation of A2A adenosine receptors enriched in the dermal surface junction (DEJ) layer promotes basal cell proliferation and differentiation, accelerates epidermal metabolism, promotes tissue regeneration, accelerates the healing and renewal of damaged skin, and repairs the underlying skin barrier. In addition, A2A adenosine receptors also participate in intercellular communication near the basement membrane, such as communication with immune cells and endothelial cells, thereby regulating the inflammatory environment within the skin.

[0004] In addition, PDRN also performs well in the field of anti-aging. PDRN exerts its anti-aging effect by regulating fibroblast activity through multiple pathways: 1) Activating the PI3KAkt and TGF-β signaling pathways to promote extracellular matrix (ECM) regeneration, which can promote the synthesis of type I and type III collagen and elastic fibers, and improve wrinkles and sagging; 2) Enhancing autophagy-lysosome and spliceosome activity to restore protein homeostasis; 3) Inhibiting chemokine signaling pathways to rebalance fibroblast-immune interactions.

[0005] After being degraded by lysing enzymes on the cell membrane in the skin, PDRN provides raw materials for the synthesis of DNA in cells by participating in the "rescue pathway," which helps to rapidly repair the DNA of damaged cells in the skin.

[0006] However, as a linear polymer of water-soluble deoxyribonucleotides, PDRN does not have an ideal ability to penetrate the stratum corneum of the skin. It is difficult for PDRN to penetrate the stratum corneum barrier and the active surface layer to enter the DEJ layer and dermis of the skin to exert its repair and anti-aging effects.

[0007] Currently, common PDRN penetration technologies in the cosmetics market include PDRN liposomes, PDRN microspheres, and PDRN combined with conventional penetration enhancers such as propylene glycol, as well as PDRN microemulsions. Other technologies include carrier nanoparticles prepared from PDRN and spermidine, and supramolecular self-assembled nanocomposites formed by the self-assembly of PDRN and mussel protein. Patent CN 121102045 A discloses a lipophilic liposome containing polydeoxyribonucleotides, its preparation method, and its application. The disclosed lipophilic liposomes, composed of lecithin and anionic / cationic emulsifiers, effectively promote transdermal absorption of PDRN and effectively resist nuclease degradation, prolonging the duration of PDRN's action in the skin. Patent CN 121270622 A discloses the extraction of polydeoxyribonucleotides and the preparation and application of PDRN-Ce microsphere clusters. It discloses that PDRN and cerium salts self-assemble through electrostatic and coordination interactions to form PDRN-Ce microsphere clusters, effectively improving penetration efficiency. Both PDRN liposomes and PDRN microspheres require relatively complex preparation processes in advance. For example, the preparation of liposomes involves complex shell membrane components, complex industrial processes, and limited encapsulation efficiency. Microsphere preparation also requires complex preparation processes such as heating and centrifugation.

[0008] Based on the mechanism of action of PDRN in repair and anti-aging, enhancing its transdermal ability will be more conducive to exerting its anti-aging and repair effects, and maximizing its therapeutic effect in practical applications. Summary of the Invention

[0009] The purpose of this invention is to overcome the shortcomings of the prior art and provide a penetration-enhancing composition, polydeoxyribonucleic acid extract and its preparation method.

[0010] To achieve the above objectives, the present invention is implemented using the following technical solution: In a first aspect, the present invention provides a method for preparing a penetration-enhancing composition, comprising: Preparation of hydroxyethyl deacetylated chitosan solution and polydeoxyribonucleotide solution; A solution of hydroxyethyl deacetylated chitosan was added to a solution of polydeoxyribonucleotides, and after stirring, isosorbide dimethyl ether was added. The mixture was stirred again until homogeneous to obtain a permeation-enhancing composition.

[0011] Furthermore, in the permeation-enhancing composition, the mass ratio of polydeoxyribonucleotides to hydroxyethyl deacetylated chitosan is 10:2~4.

[0012] Furthermore, in the permeation-enhancing composition, the mass percentage of hydroxyethyl deacetylated chitosan is 0.0003~0.12%, the mass percentage of polydeoxyribonucleotide is 0.001~0.3%, and the mass percentage of isosorbide dimethyl ether is 1~3%.

[0013] Furthermore, the preparation method of the hydroxyethyl deacetylated chitosan solution includes: Add hydroxyethyl deacetylated chitosan to deionized water, heat in a water bath, stir until completely dissolved, cool and sieve to obtain hydroxyethyl deacetylated chitosan solution; And / or, the method for preparing the polydeoxyribonucleotide solution includes: Polydeoxyribonucleotides were added to deionized water and stirred at room temperature until completely dissolved to obtain a polydeoxyribonucleotide solution.

[0014] Furthermore, the water bath heating temperature range is 45~65℃; and / or, the stirring rate at room temperature is 300~500r / min.

[0015] Furthermore, the stirring conditions include stirring continuously at a rate of 30-50 r / min for 20-30 min.

[0016] In a second aspect, the present invention also provides a penetration-enhancing composition, which is prepared by the method for preparing a penetration-enhancing composition as described in any one of the first aspects.

[0017] Thirdly, the present invention also provides a method for preparing polydeoxyribonucleotide extract, comprising: Add the thickener to deionized water, homogenize until homogeneous, heat and keep warm to obtain phase A solution; After the A phase solution is cooled, a preservative is added, and the mixture is stirred until homogeneous. The solution is then cooled again, and the penetration-enhancing composition as described in the second aspect is added. The mixture is stirred until homogeneous to obtain polydeoxyribonucleic acid extract.

[0018] Furthermore, the thickener is one or more of the following: Sphingomonas fermentation product extract, Tremella fuciformis polysaccharide, xanthan gum, Sclerotium sclerotium gum, and Carrageenanthera philoxeroides. And / or, the preservative is one or a combination of p-hydroxyacetophenone and 1,2-hexanediol.

[0019] Fourthly, the present invention also provides a method for preparing polydeoxyribonucleotide essence, wherein the heating temperature range is 75~85℃; And / or, the temperature drop gradient range is 55~65℃, and the subsequent temperature drop gradient range is 35~45℃.

[0020] Compared with the prior art, the beneficial effects achieved by the present invention are as follows: This invention utilizes the synergistic effect of hydroxyethyl deacetylated chitosan and polydeoxyribonucleic acid (PDA) aggregates, along with isosorbide dimethyl ether, to solve the transdermal penetration problem of PDAs. Isosorbide dimethyl ether enables rapid penetration through the stratum corneum barrier upon contact with the skin, facilitating initial penetration. While the hydroxyethyl deacetylated chitosan and PDA aggregates penetrate slowly during the stratum corneum penetration stage, they exhibit rapid penetration into the active layer after entering the stratum corneum. The combination of these two components allows isosorbide dimethyl ether to facilitate the rapid penetration of hydroxyethyl deacetylated chitosan and PDA aggregates through the stratum corneum, ensuring excellent penetration performance throughout the 0-12 hour period. This represents a significant breakthrough in relative penetration volume and depth, enabling PDAs to reach the DEJ layer, a key target site for PDA in the skin, regulating basal cells and subsequently activating fibroblasts in the dermis. This greatly enhances the bioavailability and anti-aging effects of PDAs in the skin.

[0021] In this invention, hydroxyethyl deacetylated chitosan and polydeoxyribonucleotides self-assemble into spheroidal aggregates based on the interaction of positive and negative charges. Hydroxyethyl deacetylated chitosan can compress the linear structure of polydeoxyribonucleotides, which helps the polydeoxyribonucleotides penetrate. The aggregates formed prevent the polydeoxyribonucleotides from being rapidly degraded by nucleases in the skin, thus achieving long-lasting and sustained-release effects.

[0022] The preparation method of the present invention does not require a complicated preparation process, but only requires simple mixing according to the steps. The composition dissolves in water to form a colorless, odorless and transparent solution with good stability and is easy to add. It can be applied to a variety of aqueous, emulsion and cream products, avoiding the application of penetration-enhancing technology due to process problems and stability issues. Attached Figure Description

[0023] Figure 1 This is a scanning electron microscope comparison diagram of the permeation-enhancing composition and polydeoxyribonucleotides prepared in Example 1 of the present invention. Figure 2 This is a schematic diagram showing the content distribution of the polydeoxyribonucleic acid essence prepared in Example 4 of the present invention at different depths of skin penetration over time. Figure 3 This is a schematic diagram showing the content distribution of the polydeoxyribonucleic acid essence prepared in Comparative Example 3 of the present invention at different depths of skin penetration over time. Figure 4 This is a schematic diagram showing the content distribution of the polydeoxyribonucleic acid essence prepared in Comparative Example 4 of the present invention at different depths of skin penetration over time. Figure 5This is a schematic diagram showing the content distribution of the polydeoxyribonucleic acid essence prepared in Example 5 of the present invention at different depths of skin penetration over time; Figure 6 This is a schematic diagram showing the relative penetration enhancement rate of the polydeoxyribonucleic acid extracts prepared in Example 4, Comparative Example 4 and Comparative Example 5 of the present invention relative to Comparative Example 3 at different time points; Figure 7 This is a schematic diagram showing the relative permeability of the polydeoxyribonucleic acid extracts prepared in Examples 4, 3, 4 and 5 of the present invention at different time points. Figure 8 This is a schematic diagram showing the penetration depth of the polydeoxyribonucleic acid extracts prepared in Examples 4, 3, 4 and 5 of the present invention at different time points. Detailed Implementation

[0024] The present invention will be further described below with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and should not be used to limit the scope of protection of the present invention.

[0025] Example 1:

[0026] This invention provides a method for preparing a penetration-enhancing composition, comprising the following steps: Add 5.0 g of hydroxyethyl deacetylated chitosan to 95 g of deionized water, heat in a water bath to 55 °C, stir at 400 r / min until completely dissolved, allow to cool naturally to room temperature, and pass through a 200 mesh sieve to obtain a hydroxyethyl deacetylated chitosan (HCS) solution.

[0027] Add 5.0g of polydeoxyribonucleic acid (PDRN) powder to 95g of deionized water and stir at 400 r / min at room temperature until it is completely dissolved to obtain a PDRN solution.

[0028] Weigh 2.0g of HCS solution and add 10g of PDRN solution. Stir continuously at 40 r / min for 25 min, then add isosorbide dimethyl ether and stir until homogeneous to obtain the penetration-enhancing composition.

[0029] A scanning electron microscope (SEM) schematic diagram of the permeation-enhancing composition and polydeoxyribonucleotides is shown below. Figure 1 As shown, isosorbide dimethyl ether has no effect on the self-assembled aggregates of PDRN and HCS.

[0030] Example 2:

[0031] The only difference between this embodiment and Example 1 is that the mass of the HCS solution is 3.0g.

[0032] Example 3:

[0033] The only difference between this embodiment and Comparative Example 1 is that the mass of the HCS solution is 4.0 g.

[0034] Comparative Example 1: The only difference between this comparative example and Example 1 is that the mass of the HCS solution is 1.5g.

[0035] Comparative Example 2: The only difference between this comparative example and Example 1 is that the mass of the HCS solution is 4.5g.

[0036] Dynamic light scattering (DLS) particle size analysis was performed on the permeation-enhancing compositions obtained in Examples 1-3 and Comparative Examples 1-2, and the results are shown in Table 1.

[0037] Table 1: Dynamic light scattering particle size analysis results of the penetration-enhancing compositions obtained in Examples 1-3 and Comparative Examples 1-2

[0038] As shown in Table 1, when the mass ratio of PDRN:HCS is between 10:2 and 4, the average hydrated particle size of the aggregates is below 300 nm, and the polydispersity index is less than 0.3. Within this mass ratio range, the size is relatively small and the particle size distribution is narrow, which is more conducive to the stable dispersion of aggregates and subsequent transdermal penetration.

[0039] Example 4:

[0040] This embodiment provides a method for preparing polydeoxyribonucleotide extract, including the following steps: First, prepare the penetration-enhancing composition: Hydroxyethyl deacetylated chitosan was added to deionized water, heated to 55°C in a water bath, and stirred at 400 r / min until completely dissolved. The solution was then allowed to cool naturally to room temperature and passed through a 200-mesh sieve to obtain a hydroxyethyl deacetylated chitosan (HCS) solution.

[0041] Polydeoxyribonucleic acid (PDRN) powder was added to deionized water and stirred at 400 r / min at room temperature until it was completely dissolved to obtain a PDRN solution.

[0042] Weigh out HCS solution and add it to PDRN solution. Stir continuously at 40 r / min for 25 min, then add isosorbide dimethyl ether and stir until homogeneous to obtain the penetration-enhancing composition.

[0043] Next, the extract of Sphingomonas fermentation product was weighed and slowly added to deionized water. The mixture was homogenized for 6 minutes until it was uniform and free of particles. The solution was then heated to 85°C and kept warm for 20 minutes to obtain phase A solution.

[0044] After the A phase solution is cooled to 60°C, add the preservatives p-hydroxyacetophenone and 1,2-hexanediol, and stir until homogeneous.

[0045] The prepared penetration-enhancing composition was added to the A-phase solution, which had been cooled to 45°C, and stirred until homogeneous to obtain PDRN essence. The preservatives p-hydroxyacetophenone and 1,2-hexanediol, along with the components of the penetration-enhancing composition, constituted the B-phase solution.

[0046] Comparative Example 3: This comparative example provides a method for preparing polydeoxyribonucleotide extract, including the following steps: Weigh out the fermentation product extract of Sphingomonas and slowly add it to deionized water. Homogenize the solution for 6 minutes until it is homogeneous and free of particles. Heat the solution to 85°C and keep it warm for 20 minutes to obtain phase A solution.

[0047] After the A phase solution is cooled to 60°C, add the preservatives p-hydroxyacetophenone and 1,2-hexanediol, and stir until homogeneous.

[0048] The PDRN solution was pre-dissolved in a small amount of deionized water and then added to the A phase solution which had been cooled to 45°C. The mixture was stirred until homogeneous to obtain the PDRN essence.

[0049] Comparative Example 4: This comparative example provides a method for preparing polydeoxyribonucleotide extract, including the following steps: Weigh out the fermentation product extract of Sphingomonas and slowly add it to deionized water. Homogenize the solution for 6 minutes until it is homogeneous and free of particles. Heat the solution to 85°C and keep it warm for 20 minutes to obtain phase A solution.

[0050] After the A phase solution is cooled to 60°C, add the preservatives p-hydroxyacetophenone and 1,2-hexanediol, and stir until homogeneous.

[0051] The HCS solution and PDRN solution were mixed and added to the A phase solution which had been cooled to 45°C. The mixture was stirred until homogeneous to obtain PDRN essence.

[0052] Comparative Example 5: This comparative example provides a method for preparing polydeoxyribonucleotide extract, including the following steps: Weigh out the fermentation product extract of Sphingomonas and slowly add it to deionized water. Homogenize the solution for 6 minutes until it is homogeneous and free of particles. Heat the solution to 85°C and keep it warm for 20 minutes to obtain phase A solution.

[0053] After the A phase solution is cooled to 60°C, add the preservatives p-hydroxyacetophenone and 1,2-hexanediol, and stir until homogeneous.

[0054] PDRN solution and isosorbide dimethyl ether were added to phase A solution which had been cooled to 45°C and stirred until homogeneous to obtain PDRN essence.

[0055] Comparative Example 6: Weigh out the fermentation product extract of Sphingomonas and slowly add it to deionized water. Homogenize the solution for 6 minutes until it is homogeneous and free of particles. Heat the solution to 85°C and keep it warm for 20 minutes to obtain phase A solution.

[0056] After the A phase solution is cooled to 60°C, add the preservatives p-hydroxyacetophenone and 1,2-hexanediol, and stir until homogeneous.

[0057] Tetrahydropiperine, isosorbide dimethyl ether, and a small amount of deionized water were mixed and pre-dissolved, and then added to phase A solution which had been cooled to 45°C. The mixture was stirred until homogeneous to obtain PDRN essence.

[0058] Examples 5-7 were prepared using the same method as in Example 4. The specific component ratios of the PDRN essences obtained in Examples 5-7 and Comparative Examples 3-7 are shown in Table 2. In Table 2, the preservative p-hydroxyacetophenone, 1,2-hexanediol, and the components of the penetration-enhancing composition are used as phase B solutions.

[0059] Table 2: Component ratios of PDRN extracts obtained in Examples 4-7 and Comparative Examples 4-7

[0060] The performance of the PDRN extracts prepared in Examples 4-7 and Comparative Examples 3-6 is analyzed below.

[0061] I. Stability Testing The PDRN extracts prepared in Examples 4-7 and Comparative Examples 3-6 were all aliquoted into transparent containers. The transparent containers were stored at -18℃, 4℃, 25℃, and 45℃ in the dark and at room temperature under light conditions to track stability. The appearance, odor, and viscosity changes of the samples were observed after 1 month and 3 months to evaluate stability. The stability results are shown in Table 3.

[0062] Table 3: Stability test results of PDRN extracts prepared in Examples 4-7 and Comparative Examples 3-6

[0063] As can be seen from the data in Table 3, Comparative Example 4 was unstable, and an oily substance precipitated on the surface of the liquid and the wall of the transparent container after placement.

[0064] II. Ex vivo skin permeability test Test Methods: The transdermal penetration characteristics of different PDRN essences were evaluated using an ex vivo piglet skin model. Skin from the back of piglets was used to prepare a 2 cm × 2 cm test area. The following samples were applied: Example 4, Comparative Example 3, Comparative Example 4, and Comparative Example 5. Depth Raman imaging was performed at 0, 2, 4, 8, 10, and 12 h after application using a LabRAM Odyssey high-resolution confocal microscopy system (HORIBA). The scanning longitudinal step size was 10 μm, and the area was 20 μm × 120 μm. By analyzing the relative intensity of the characteristic Raman peaks (distinct from the intrinsic skin signal) at different depths, the relative permeability (normalized percentage change of characteristic peaks) and maximum penetration depth of PDRN at each time point were calculated.

[0065] Test results: like Figures 2-6 As shown, the 0.2% PDRN in the serum of Comparative Example 3 penetrated slowly, with a relative penetration rate of 0.54% after 4 hours and only 4.86% after 12 hours, with a maximum depth of 40 μm.

[0066] After adding 0.06% HCS, the penetration of the essence in Comparative Example 4 was significantly improved: the relative penetration rate was 3.21% at 4 hours and 6.93% at 12 hours, with a depth of 80 μm. The relative penetration improvement rate at 4 hours was as high as 494.44%, indicating that HCS has a good penetration-promoting effect.

[0067] After adding 2% isosorbide dimethyl ether (penetration enhancer), the relative penetration rate of the Comparative Example 5 essence was 2.85% after 4 hours, and increased to 7.85% after 12 hours, with a depth of 80 μm. The relative penetration improvement rate after 4 hours was 427.8%.

[0068] The essence in Example 4, which included 0.06% HCS and 2% isosorbide dimethyl ether, exhibited the best penetration performance, with a relative penetration rate of 6.28% at 4 hours and 9.33% at 12 hours, reaching a depth of 110 μm (penetrating the dermis). The relative penetration improvement rate at 4 hours was 1063.0%.

[0069] The results showed that both HCS and isosorbide dimethyl ether can effectively promote the transdermal penetration of PDRN. The combined application of the two has a significant synergistic effect on penetration depth and relative permeability, which is beneficial to improving the skin bioavailability of PDRN.

[0070] Combination Figure 7 and Figure 8By observing the penetration depth and relative permeability at different time points, it can be seen that the initial 2-2.5 days after application to the skin... The serum containing 2% isosorbide dimethyl ether (2%) showed faster and greater PDRN penetration, with significantly better penetration depth and relative permeability than the serum without 2% isosorbide dimethyl ether. This indicates that 2% isosorbide dimethyl ether can rapidly promote penetration in the early stages of skin contact, helping PDRN penetrate the stratum corneum. The serum containing 0.06% HCS showed a significant improvement in relative permeability and penetration depth after 4 hours, even surpassing the best-performing serum with 2% isosorbide dimethyl ether. Then, at 8 hours, the penetration depth of the serum containing 0.06% HCS was approximately twice that of the serum containing 2% isosorbide dimethyl ether. This demonstrates that 2% isosorbide dimethyl ether can quickly penetrate the stratum corneum in the early stages, and due to its amphiphilic properties, it maintains relatively stable penetration performance even after entering the active layer of the skin. In contrast, 0.06% HCS showed a rapid increase in relative penetration amount and depth after penetrating the stratum corneum, penetrating the skin faster and exhibiting excellent penetration performance in the later stages.

[0071] When the two are combined, the overall penetration performance is excellent from the early to the late stages. The relative penetration amount and penetration depth are far superior to the essence of 2% isosorbide dimethyl ether and 0.06% HCS alone. The penetration depth reaches 110µm in 12 hours, reaching the dermal layer of isolated pig skin.

[0072] Comparing the penetration enhancement rates of PDRN serums with added penetration enhancers compared to those without, we found that: during the period of 4h to 8h, all formulations containing penetration enhancers significantly improved penetration rates, especially the combined use group, where the improvement rate exceeded 1000% at 4h; at 10h and 12h, the improvement rate tended to level off, but still maintained a positive increase; at 2h, the control group had a penetration rate of 0%, so the improvement rate could not be calculated, but the absolute penetration amount showed that the group containing penetration enhancers had begun to penetrate (0.78%~1.31%). III. Patch Test Test Method: Thirty healthy volunteers meeting the inclusion criteria were selected (it is recommended that those with sensitive skin or mildly impaired skin barrier be included), excluding those with a history of using anti-inflammatory drugs or allergies within the past week. Example 4 was used as the test sample, and physiological saline was used as a negative control. Approximately 0.020-0.025 mL of the essence from Example 4 was dropped into the patch applicator chamber, and an equal volume of physiological saline was added to the control chamber. The patch applicator was applied to the normal skin on the back (or flexor surface of the forearm) of the subject and secured with non-irritating tape, remaining in place for 48 hours. After removing the patch applicator, skin reactions were observed at 0.5 hours (immediately after removal), 24 hours, and 48 hours, referring to the grading standards for skin occlusive patch tests in the *Cosmetic Safety Technical Specifications*.

[0073] Interpretation criteria: The following grading criteria for skin occlusive patch tests are used for scoring, as per the "Cosmetic Safety Technical Specifications": Grade 0: Negative reaction, no reaction whatsoever.

[0074] Grade 1: Suspicious reaction, with only slight erythema.

[0075] Grade 2: Weak positive reaction (erythema, infiltration, papules, no vesicles).

[0076] Grade 3: Strong positive reaction (erythema, infiltration, papules, vesicles).

[0077] Grade 4: Extremely positive reaction (severe redness, large blisters, erosion).

[0078] The test results are shown in Table 4 (Note: Level 1 is a suspicious reaction and is not counted as positive).

[0079] Table 4: Patch test results of the essence obtained in Example 4

[0080] As shown in Table 4, under the experimental conditions, no positive reactions of grade 2 or higher occurred in any of the 30 subjects. The skin occlusive patch test result of the essence in Example 4 was negative, indicating that it has no obvious skin irritation or sensitization, and has good safety and tolerability when applied to human skin.

[0081] IV. Human Body Moisturizing, Repairing, and Anti-Wrinkle Efficacy Test Thirteen subjects aged 28-55 years were selected. The control group and the sample group were used on the left and right sides of the subjects' faces, respectively. The products were applied twice a day during the normal skin care routine in the morning and evening for 28 days. Skin physiological indicators were tested before use and on the 14th and 28th days of use. Before the test, the subjects cleaned and dried their faces. After sitting quietly for 30 minutes in a constant temperature and humidity environment (21±1℃, 50±10%), skin indicators were tested on the left and right sides of the face. Elasticity and firmness were tested using a Cutometer®, transepidermal water loss was tested using a Tewameter®™ HEX probe, and stratum corneum moisture content was tested using a Corneometer® CM825.

[0082] The 0.2% PDRN essence of Comparative Example 5 was used as the control group, and the 0.2% combined penetration-enhancing PDRN essence (containing 0.06% HCS + 2% isosorbide dimethyl ether) of Example 4 was used as the sample group.

[0083] The test results are shown in Table 5.

[0084] Table 5: Human efficacy test results of the extracts obtained from Comparative Example 5 and Example 4

[0085] As shown in Table 5, the improvement rates of firmness and transepidermal water loss (TEWL) were negative. The larger the absolute value, the better the improvement effect. Both the control group and the sample group showed significant improvement in the four tested indicators. However, the sample group, which used the essence with the combined penetration-enhancing technology, showed more significant improvement in elasticity, firmness, and transepidermal water loss after 28 days of use. This indicates that the essence in the sample has better anti-wrinkle, firming, and repairing effects.

[0086] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method of making a penetration enhancing composition, characterized by, include: Preparation of hydroxyethyl deacetylated chitosan solution and polydeoxyribonucleotide solution; A solution of hydroxyethyl deacetylated chitosan was added to a solution of polydeoxyribonucleotides, and after stirring, isosorbide dimethyl ether was added. The mixture was stirred again until homogeneous to obtain a permeation-enhancing composition.

2. The method of claim 1, wherein the permeation enhancing composition is prepared by, In the permeation-enhancing composition, the mass ratio of polydeoxyribonucleotides to hydroxyethyl deacetylated chitosan is 10:2~4.

3. The method for preparing the penetration-enhancing composition according to claim 1, characterized in that, In the permeation-enhancing composition, the mass percentage of hydroxyethyl deacetylated chitosan is 0.0003~0.12%, the mass percentage of polydeoxyribonucleotide is 0.001~0.3%, and the mass percentage of isosorbide dimethyl ether is 1~3%.

4. The method for preparing the penetration-enhancing composition according to claim 1, characterized in that, The preparation method of the hydroxyethyl deacetylated chitosan solution includes: Add hydroxyethyl deacetylated chitosan to deionized water, heat in a water bath, stir until completely dissolved, cool and sieve to obtain hydroxyethyl deacetylated chitosan solution; And / or, the method for preparing the polydeoxyribonucleotide solution includes: Polydeoxyribonucleotides were added to deionized water and stirred at room temperature until completely dissolved to obtain a polydeoxyribonucleotide solution.

5. The method for preparing the penetration-enhancing composition according to claim 4, characterized in that, The water bath heating temperature range is 45~65℃; and / or, the stirring rate at room temperature is 300~500r / min.

6. The method for preparing the penetration-enhancing composition according to claim 1, characterized in that, The stirring conditions include stirring continuously at a rate of 30-50 r / min for 20-30 min.

7. A penetration-enhancing composition, characterized in that, It is prepared by the method for preparing the permeation-enhancing composition according to any one of claims 1 to 6.

8. A method for preparing polydeoxyribonucleotide extract, characterized in that, include: Add the thickener to deionized water, homogenize until homogeneous, heat and keep warm to obtain phase A solution; After the A phase solution cools down, add the preservative, stir until homogeneous, cool down again, add the penetration-enhancing composition as described in claim 7, and stir until homogeneous to obtain polydeoxyribonucleic acid extract.

9. The method for preparing polydeoxyribonucleotide extract according to claim 8, characterized in that, The thickener is one or more of the following: Sphingomonas fermentation product extract, Tremella fuciformis polysaccharide, xanthan gum, Sclerotium sclerotium gum, and Carrageenanthe javanica. And / or, the preservative is one or a combination of p-hydroxyacetophenone and 1,2-hexanediol.

10. The method for preparing polydeoxyribonucleotide extract according to claim 8, characterized in that, The heating temperature range is 75~85℃; And / or, the temperature drop gradient range is 55~65℃, and the subsequent temperature drop gradient range is 35~45℃.

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