Macromolecule delivery system containing PDRN / DNA sodium, collagen and polypeptide, preparation method and application

Abstract

The invention provides a macromolecular delivery system containing PDRN / DNA sodium, collagen and polypeptide, and a preparation method and application thereof, and belongs to the technical field of skin care products. The macromolecular delivery system comprises a penetration enhancer and an active matter; the penetration enhancer is prepared from hydroxyethyl chitosan and inositol. The macromolecular delivery system disclosed by the invention has a very good promotion effect on transdermal absorption of DNA sodium, collagen or a combination thereof, the cumulative permeation amount per unit area is greatly increased, and the DNA sodium, the collagen or the combination thereof is deepest delivered to a corium layer.

Description

Technical Field

[0001] This invention provides a macromolecular delivery system, preparation method, and application comprising sodium PDRN / DNA, collagen, and peptides, belonging to the field of skincare technology. Background Technology

[0002] Collagen and sodium DNA have permeability barriers during transdermal absorption: Collagen, as a large protein molecule with a molecular weight of 30kDa-300kDa, far exceeds the passive diffusion threshold of the lipid bilayer of the stratum corneum, making it difficult to penetrate the intact stratum corneum into the deep epidermis or dermis; similarly, large molecular weight sodium DNA is blocked by the stratum corneum barrier and can only play a role in the skin surface or superficial epidermis.

[0003] Chinese patent CN116496429B discloses a self-assembled transdermal delivery system based on hydroxyethyl deacetylated chitosan, its preparation, and its application. The system is prepared using chitin, 1,2-epoxybutane, and ethylene oxide as main raw materials. The resulting hydroxyethyl deacetylated chitosan has an average molecular weight of 7460 Da and is used as a penetration enhancer for active ingredients such as peptides, proteins, PCA, Zn, and ergothioneine. However, experiments have shown that while it can improve the transdermal absorption rate of collagen and sodium DNA to some extent when used alone, its effect is limited and requires further improvement.

[0004] Chinese patent CN119345066A discloses a penetration-enhancing composition and its application in skincare products. The composition is a compound of hydroxyethyl deacetylated chitosan and decapeptide-4, wherein: 0.015-0.15 parts of hydroxyethyl deacetylated chitosan; and 0.0001-0.0006 parts of decapeptide-4. However, experiments have shown that while the compound of hydroxyethyl deacetylated chitosan and decapeptide-4 can improve the transdermal absorption rate of collagen and sodium DNase to some extent, the effect is limited and needs further improvement. Summary of the Invention

[0005] This invention provides a macromolecular delivery system comprising sodium PDRN / DNA, collagen, and peptides, a preparation method, and applications. The macromolecular delivery system can enhance the permeation and absorption effects of sodium PDRN / DNA, collagen, and peptides.

[0006] In a first aspect, the present invention provides: a macromolecular delivery system comprising sodium PDRN / DNA, collagen, and polypeptides, including a penetration enhancer and an active ingredient; The penetration enhancer includes hydroxyethyl deacetylated chitosan and inositol.

[0007] Furthermore, the macromolecular delivery system includes a penetration enhancer and an active ingredient in a mass ratio of (1.1-3.6):(0.0003-90).

[0008] Preferably, the macromolecular delivery system comprises a penetration enhancer and an active ingredient in a mass ratio of (1.1-3.6):(0.7-90).

[0009] More preferably, the macromolecule delivery system comprises a penetration enhancer and an active ingredient in a mass ratio of 1.1:07, 1.2:1, 1.5:10, 1.8:20, 2:25, 2.2:35, 2.6:45, 3:55, 3.2:65, 3.4:75, 3.6:85, 3.6:90, or any combination thereof.

[0010] Furthermore, the penetration enhancer comprises hydroxyethyl deacetylated chitosan and inositol in a mass ratio of (0.5-3):(0.0001-0.6).

[0011] Preferably, the penetration enhancer comprises hydroxyethyl deacetylated chitosan and inositol in a mass ratio of (2-3):(0.0001-0.3).

[0012] More preferably, the penetration enhancer comprises hydroxyethyl deacetylated chitosan and inositol in a mass ratio of 2:0.0001, 2:0.1, 2:0.2, 2:0.3, 2.5:0.005, 3:0.05 or any combination thereof.

[0013] Furthermore, the active ingredient includes at least one of PDRN and collagen.

[0014] Furthermore, the collagen includes at least one of recombinant collagen, natural collagen, and soluble collagen.

[0015] Furthermore, the macromolecule delivery system also includes water.

[0016] Secondly, the present invention provides: a method for preparing a macromolecule delivery system, comprising the following steps: (1) Hydroxyethyl deacetylated chitosan and inositol were mixed evenly to obtain a penetration enhancer; (2) Mix the penetration enhancer and active ingredient evenly to obtain the product.

[0017] Furthermore, the preparation method further includes: adding the penetration enhancer and the active ingredient to water and dispersing them evenly.

[0018] Thirdly, the present invention provides a cosmetic product comprising the aforementioned macromolecule delivery system.

[0019] Furthermore, the dosage form of the cosmetic includes at least one of solution, lotion, cream, spray and mask.

[0020] Fourthly, the present invention provides the application of a macromolecular delivery system in the preparation of cosmetics.

[0021] Furthermore, the application includes improving the transdermal absorption rate of the active ingredient.

[0022] The beneficial effects of this invention are as follows: (1) The present invention uses hydroxyethyl deacetylated chitosan and inositol as a combined permeation enhancer, which has a synergistic effect and promotes the transdermal absorption of sodium DNA, collagen, soluble collagen or their combination. Compared with comparative examples 1-5, the cumulative permeation per unit area is increased by 20-68%, reaching the deepest layer of the dermis.

[0023] (2) Hydroxyethyl deacetylated chitosan can disrupt the orderly arrangement of the lipid bilayer by interacting with the intercellular lipids of the stratum corneum, thereby increasing the hydrophilic channels of the stratum corneum; while inositol, as a hexahydrol, can soften the stratum corneum through hydration and reduce the density of keratinocytes. The two work together to broaden the transdermal pathway of the active ingredients in a specific dosage relationship. Detailed Implementation

[0024] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0025] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention.

[0026] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0027] The reagents used in this invention are described below: The inositol used in this invention has CAS number 87-89-8.

[0028] The recombinant collagen used in this invention is recombinant human collagen type I, 50±3 kDa.

[0029] The PDRN mentioned in this invention is sodium DNA, CAS number: 9007-49-2.

[0030] Soluble collagen is: collagen oligopeptide grade, 90% purity, CAS number: 9007-34-5.

[0031] The raw materials and preparation methods used in this invention comply with the relevant provisions of the "Cosmetic Safety Technical Specifications".

[0032] I. A macromolecular delivery system comprising sodium PDRN / DNA, collagen, and peptides The material amounts of the macromolecular delivery systems containing sodium PDRN / DNA, collagen, and peptides in Examples 1-5 and Comparative Examples 1-5 are shown in Tables 1 and 2.

[0033] Table 1 (Unit: wt%)

[0034] Table 2 (Unit: wt%)

[0035] Note: * indicates that inositol is replaced with decapeptide-4; # indicates that hydroxyethyl deacetylated chitosan is replaced with carboxymethylated chitosan.

[0036] II. Preparation Method 1. Preparation method of hydroxyethyl deacetylated chitosan: It was prepared by the preparation method described in Example 1 of CN116496429B.

[0037] 2. The preparation methods of Examples 1-5 and Comparative Examples 1-5 are as follows: the penetration enhancer and active ingredient are added to water and dispersed evenly.

[0038] III. Effect Test 1. Transdermal absorption effect experiment The experiment was conducted using a diffusion cell as described in GB / T 27818-2011, "In vitro test methods for skin absorption of chemicals". Experimental samples: macromolecule delivery systems of Examples 1-5 and Comparative Examples 1-5.

[0039] Experimental materials: detached pigskin from the back of a miniature pig. Specifically, after the Bama miniature pig was euthanized, the hair on its abdominal skin was quickly removed using an electric hair clipper; the hairless skin was peeled off, and the subcutaneous tissue was thoroughly removed; the treated skin was rinsed with physiological saline; the skin was soaked in physiological saline for 30 minutes; the skin was removed from the physiological saline, and its surface moisture was gently blotted dry with filter paper before proceeding with subsequent experiments.

[0040] Experimental Methods: The Franz vertical diffusion cell-LC-MS method was used to fix the prepared ex vivo skin between the drug delivery cell and the receiving cell. The effective permeation area of ​​the diffusion cell was 2.80 cm². 2The stratum corneum faces the supply chamber, while the dermis side contacts the receiving solution to remove air bubbles. The receiving solution is physiological saline. Accurately weigh 0.1g each of Examples 1-5 and Comparative Examples 1-5 and apply them to the skin, ensuring close contact. Place a small glass bead in the receiving chamber to help agitate the receiving solution. Seal the supply chamber with plastic wrap to prevent moisture evaporation. Maintain a constant temperature of 37±0.5℃ using a water bath outside the receiving chamber.

[0041] After 12 hours, all the receiving liquid was collected, centrifuged, and the supernatant was filtered through a microporous membrane. The total concentration of the active ingredient in the filtrate was determined by liquid chromatography-tandem mass spectrometry, and the cumulative permeation per unit area Q was calculated.

[0042] Among them, C n V represents the total concentration of active material measured at the nth sampling point; V is the volume of the receiving cell; and S is the effective diffusion area.

[0043] 2. Fluorescent labeling experiment (1) Examples 1-5 and Comparative Examples 1-5 were pre-loaded with the fluorescent probe FITC: 1g of recombinant collagen was dissolved in 0.1M pH 5.5 buffer solution, and then 0.1g of fluorescent probe FITC was added. After reacting for 6 hours, unreacted FITC was removed by dialysis, and the collagen was obtained by freeze-drying.

[0044] 1g of soluble collagen was dissolved in 0.1M pH 5.5 buffer solution, and then 0.1g of fluorescent probe FITC was added. After reacting for 6 hours, unreacted FITC was removed by dialysis, and the soluble collagen was obtained by freeze-drying.

[0045] Dissolve 1g of sodium DNA in 0.1M pH 5.5 buffer solution, then add 0.1g of fluorescent probe FITC. After reacting for 6 hours, remove unreacted FITC by dialysis, and freeze-dry to obtain fluorescently labeled sodium DNA.

[0046] Fluorescently labeled raw materials were used to prepare Examples 1-5 and Comparative Examples 1-5 according to Table 1.

[0047] (3) Experimental methods Rats were shaved off their back hair and randomly divided into groups of 3 rats each: a blank group, Example 1-5 groups, and Comparative Example 1-5 groups.

[0048] The prepared formulations of Examples 1-5 and Comparative Examples 1-5 were applied to the dorsal skin of rats at a concentration of 0.1 g / cm³. 2The rats were treated for 8 hours, and the control group was treated with an equal amount of water. Then, the rats were euthanized by cervical dislocation, and the skin on their backs was surgically removed, washed with PBS, and air-dried. Sections of the removed skin were embedded in paraffin (5µm thickness), fixed on glass slides, and subjected to λ... ex Observation was performed under CLSM using an argon laser beam excited at 488 nm, while fluorescence emission was observed at λ. em The fluorescence intensity was measured at 525 nm, and the deepest part of the fluorescent layer was observed, along with the relative fluorescence intensity of the deepest part (with the blank group as the reference).

[0049] 3. Experimental Results The results of the transdermal absorption rate experiment and the fluorescent labeling experiment are shown in Table 3.

[0050] Table 3

[0051] The comparison between Example 2 and Comparative Examples 1-5 demonstrates that the combined use of hydroxyethyl deacetylated chitosan and inositol as a penetration enhancer has a synergistic effect, which greatly promotes the transdermal absorption of sodium DNA, collagen, or combinations thereof. Compared with Comparative Examples 1-5, the cumulative penetration per unit area is increased by 20-68%, reaching as deep as the dermis.

[0052] However, replacing the types of hydroxyethyl deacetylated chitosan and inositol, or changing their dosage, will disrupt this synergistic effect. As seen in Comparative Examples 1-5, whether only hydroxyethyl deacetylated chitosan or inositol is used, or the types or dosages are changed, the cumulative penetration per unit area decreases and the relative fluorescence intensity decreases. In fact, Comparative Examples 2 and 5 cannot even reach the dermis.

[0053] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A macromolecular delivery system comprising PDRN / DNA sodium class, collagen and polypeptide, characterized in that, Including penetration enhancers and active ingredients; The penetration enhancer includes hydroxyethyl deacetylated chitosan and inositol.

2. The macromolecule delivery system of claim 1, wherein, It includes penetration enhancers and active ingredients in a mass ratio of (1.1-3.6):(0.0003-90).

3. The macromolecule delivery system according to claim 1, characterized in that, The penetration enhancer comprises hydroxyethyl deacetylated chitosan and inositol in a mass ratio of (0.5-3):(0.0001-0.6).

4. The macromolecule delivery system according to claim 3, characterized in that, The penetration enhancer comprises hydroxyethyl deacetylated chitosan and inositol in a mass ratio of (2-3):(0.0001-0.3).

5. The macromolecule delivery system according to claim 1, characterized in that, The active ingredient includes at least one of PDRN and collagen.

6. The macromolecule delivery system according to claim 5, characterized in that, The collagen includes at least one of recombinant collagen, natural collagen, and soluble collagen.

7. A method for preparing the macromolecule delivery system according to any one of claims 1-6, characterized in that, Includes the following steps: (1) Hydroxyethyl deacetylated chitosan and inositol were mixed evenly to obtain a penetration enhancer; (2) Mix the penetration enhancer and active ingredient evenly to obtain the product.

8. A cosmetic product, characterized in that, Includes the macromolecule delivery system according to any one of claims 1-6.

9. The use of the macromolecular delivery system according to any one of claims 1-6 in the preparation of cosmetics.

10. The application according to claim 9, characterized in that, This includes improving the transdermal absorption rate of active ingredients.

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