A medical aesthetic active ingredient composition having a repairing anti-aging effect

Abstract

This invention relates to the field of topical medical aesthetic active ingredients, and discloses a medical aesthetic active ingredient composition with repairing and anti-aging effects. The composition, by weight, comprises bilayer unpacking polydeoxyribonucleotide complex particles, ectoine, ergothioneine, panthenol, allantoin, and an aqueous carrier. The bilayer unpacking polydeoxyribonucleotide complex particles include a ternary core formed by polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide; an enzyme-sensitive shell consisting of low-molecular-weight sodium hyaluronate and trehalose located outside the ternary core; and a discontinuous lipid interlayer formed by hydrogenated lecithin, ceramide NP, and cholesterol. This composition can reduce the free proportion of polydeoxyribonucleotides in an aqueous system and exhibit detectable particle size and release changes in the presence of hyaluronidase, making it suitable for topical repairing and anti-aging ingredient systems.

Description

Technical Field

[0001] This invention relates to the field of topical medical aesthetic active ingredients, specifically to a medical aesthetic active ingredient composition with repair and anti-aging effects. Background Technology

[0002] With the development of topical repair and anti-aging products, active ingredients such as polydeoxyribonucleotides, hyaluronic acid, carnosine, panthenol, allantoin, ectoine, ergothioneine, and ceramides have been widely used in topical serums, gels, facial masks, and lyophilized reconstituted ingredients. Among these, polydeoxyribonucleotides are typically used as nucleotide-based active ingredients, hyaluronic acid and its salts are commonly used for moisturizing and film formation, ceramide lipids are often used for barrier care, and carnosine and ergothioneine are frequently used for anti-oxidative stress care. Existing technologies also include compounding or encapsulating polydeoxyribonucleotides with hyaluronic acid, peptides, chitosan, or liposomes to improve their dispersibility and stability in topical systems.

[0003] However, existing topical medical aesthetic active ingredient compositions mostly employ direct compounding of active ingredients, monolayer coating, or physical mixing with liposomes. In these systems, polydeoxyribonucleotides tend to exist in a free state in the aqueous carrier, resulting in a high early release rate. Hyaluronic acid materials, chitosan oligosaccharide materials, and lipid components typically lack a clear layered binding relationship, and lipid components easily form free liposomes or lipid aggregates, making it difficult to stably participate in the regulation of the outer structure of polydeoxyribonucleotides. At the same time, although ordinary hyaluronic acid coated particles can form a certain coating structure, it is difficult to simultaneously achieve both a low burst release state in the absence of enzymes and a responsive release in the presence of hyaluronidase.

[0004] Therefore, existing technologies still require a topical medical aesthetic active ingredient composition that allows polydeoxyribonucleotides to be primarily in an encapsulated state, and reduces early free release in the aqueous system through layered synergy between the ternary core, enzyme-sensitive shell, and discontinuous lipid interlayer. Simultaneously, it should form detectable particle size and release changes in the presence of hyaluronidase, thereby improving the structural stability and quality control of the topical medical aesthetic active ingredient composition. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the present invention provides a medical aesthetic active ingredient composition with repair and anti-aging effects to solve the technical problems existing in the prior art.

[0006] The above-mentioned technical objective of the present invention is achieved through the following technical solution: A medical aesthetic active ingredient composition with repair and anti-aging effects includes 0.5-8 parts of double-layer unpacking type polydeoxyribonucleotide complex particles, 0.2-4 parts of osmotic pressure protection component, 0.01-1 parts of antioxidant stress component, 0.05-3 parts of soothing and repairing component, and an aqueous phase carrier, with the aqueous phase carrier supplemented to 100 parts; Osmotic protection components include ectoine, antioxidant stress components include ergothioneine, and soothing and repairing components include panthenol and allantoin. The double-layer unpacking polydeoxyribonucleic acid complex particle includes a ternary core, an enzyme-sensitive shell, and a discontinuous lipid interlayer; The ternary core is formed by electrostatic recombination of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide. The enzyme-sensitive shell consists of low-molecular-weight sodium hyaluronate and trehalose. The low-molecular-weight sodium hyaluronate is located outside the ternary core, and the trehalose is dispersed in the shell formed by the low-molecular-weight sodium hyaluronate. The discontinuous lipid block includes hydrogenated lecithin, ceramide NP and cholesterol. The discontinuous lipid block is composed of multiple sheet lipid regions spaced apart from each other. The sheet lipid regions are distributed on the outside of the enzyme-sensitive shell, and at least some of the sheet lipid regions are embedded in the enzyme-sensitive shell.

[0007] Preferably, based on a total mass of 100 parts for the double-layer unpacked polydeoxyribonucleic acid complex particles, the ternary core comprises 20-55 parts, the enzyme-sensitive shell comprises 30-65 parts, and the discontinuous lipid intercalation comprises 5-20 parts, and the sum of the mass parts of the ternary core, the enzyme-sensitive shell, and the discontinuous lipid intercalation comprises 100 parts.

[0008] Preferably, in the ternary core, the mass ratio of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide is 1:0.05-0.4:0.2-1.0; The length of the polydeoxyribonucleotide fragment is 80-600 bp, and the weight-average molecular weight of the quaternized chitosan oligosaccharide is 1-10 kDa.

[0009] Preferably, in the enzyme-sensitive shell, the mass ratio of low molecular weight sodium hyaluronate to trehalose is 1:0.3-2.5; The weight-average molecular weight of the low molecular weight sodium hyaluronate is 20-150 kDa.

[0010] Preferably, in the discontinuous lipid block, the mass ratio of hydrogenated lecithin, ceramide NP, and cholesterol is 1:0.03-0.25:0.08-0.60; The average sheet thickness of the lipid sheet region was measured to be 5-40 nm using transmission electron microscopy.

[0011] Preferably, the double-layer unpacking polydeoxyribonucleotide composite particles have a D50 particle size of 80-350 nm, a polydispersity index of no more than 0.35, a Zeta potential of -20 mV to +10 mV, and a polydeoxyribonucleotide loading rate of no less than 70%.

[0012] Preferably, after being placed in an aqueous system at 37°C, pH 5.0-6.5, and free of hyaluronidase for 4 hours, the polydeoxyribonucleic acid release rate of the double-layer unpacking polydeoxyribonucleic acid composite particles is no higher than 30%. After being placed in an aqueous system at 37℃, pH 5.0-6.5, and hyaluronidase concentration of 10-100 U / mL for 24 hours, the polydeoxyribonucleotide release rate of the double-layer unpacking polydeoxyribonucleotide composite particles was 55%-85%. The release rate is calculated as the ratio of the mass of polydeoxyribonucleic acid (PDA) in the release medium to the initial mass of PDA in the bilayer unpacked PDA composite particles.

[0013] Preferably, after being placed in an aqueous system at 37°C, pH 5.0-6.5, and hyaluronidase concentration of 10-100 U / mL for 2 hours, the D50 particle size of the double-layer unpacked polydeoxyribonucleic acid composite particles is 1.2-2.5 times the D50 particle size before placement.

[0014] Preferably, the mass ratio of panthenol to allantoin is 1:0.02-0.3; Ectoin, ergothioneine, panthenol, and allantoin are all dispersed in an aqueous carrier.

[0015] Preferably, the aqueous support comprises purified water, glycerol, and 1,3-propanediol, wherein the total mass of glycerol and 1,3-propanediol is 2%-12% of the mass of the aqueous support. The pH of the medical aesthetic active ingredient composition is 5.0-6.5, the total content of polydeoxyribonucleotides in the medical aesthetic active ingredient composition is 0.005wt%-0.8wt%, and the mass of polydeoxyribonucleotides not encapsulated in the ternary core accounts for no more than 20% of the total mass of polydeoxyribonucleotides.

[0016] In summary, the present invention has the following main beneficial effects: By first constructing a ternary core of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide, and then forming an enzyme-sensitive shell composed of low-molecular-weight sodium hyaluronate and trehalose on the outside of the ternary core, the polydeoxyribonucleotides are mainly in a loaded state, reducing their free proportion in the aqueous carrier. This achieves the effect of improving the dispersion stability of polydeoxyribonucleotides in topical medical aesthetic active ingredient systems and reducing the risk of early rapid release.

[0017] By setting a discontinuous lipid interlayer composed of hydrogenated lecithin, ceramide NP and cholesterol on the outside of the enzyme-sensitive shell, and partially embedding multiple spaced-apart sheet lipid regions into the enzyme-sensitive shell, the lipid components no longer exist in the form of ordinary liposomes or simple physical mixtures, but form a layered composite structure with the enzyme-sensitive shell. This achieves the effects of improving the stability of the outer layer structure of the composite particles, enhancing particle retention in the topical system and reducing the burst release of polydeoxyribonucleotides.

[0018] By limiting the low release state of the composite particles in the absence of hyaluronidase, the release range in the presence of hyaluronidase, and the particle size change factor after hyaluronidase treatment, a detectable correspondence is established between the loosening process of the enzyme-sensitive shell, the release process of polydeoxyribonucleotides, and the changes in particle structure. This achieves the effect of distinguishing it from ordinary PDRN hyaluronic acid coated particles, chitosan oligosaccharide hyaluronic acid aggregated particles, and ceramide liposome physical mixing systems. It also helps to provide a clear basis for the stable preparation and quality control of external repair and anti-aging raw materials. Attached Figure Description

[0019] Figure 1 This is a flowchart of the method of the present invention. Detailed Implementation

[0020] 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.

[0021] Example 1 refer to Figure 1 A medical aesthetic active ingredient composition with repair and anti-aging effects includes 0.5-8 parts of double-layer unpacking type polydeoxyribonucleotide complex particles, 0.2-4 parts of osmotic pressure protection component, 0.01-1 parts of antioxidant stress component, 0.05-3 parts of soothing and repairing component, and an aqueous phase carrier, with the aqueous phase carrier supplemented to 100 parts; Osmotic protection components include ectoine, antioxidant stress components include ergothioneine, and soothing and repairing components include panthenol and allantoin. The double-layer unpacking polydeoxyribonucleic acid complex particle includes a ternary core, an enzyme-sensitive shell, and a discontinuous lipid interlayer; The ternary core is formed by electrostatic recombination of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide. The enzyme-sensitive shell consists of low-molecular-weight sodium hyaluronate and trehalose. The low-molecular-weight sodium hyaluronate is located outside the ternary core, and the trehalose is dispersed in the shell formed by the low-molecular-weight sodium hyaluronate. The discontinuous lipid block includes hydrogenated lecithin, ceramide NP and cholesterol. The discontinuous lipid block is composed of multiple sheet lipid regions spaced apart from each other. The sheet lipid regions are distributed on the outside of the enzyme-sensitive shell, and at least some of the sheet lipid regions are embedded in the enzyme-sensitive shell.

[0022] This composition is intended for use in topical skincare products, topical repair serums, topical gels, topical face mask liquids, or lyophilized reconstituted topical ingredients, and is not intended for injection, implantation, or deep wound administration.

[0023] The topical medical aesthetic active ingredient composition, by weight, comprises 0.5-8 parts of bilayer unpacking polydeoxyribonucleotide complex particles, 0.2-4 parts of osmotic pressure protection component, 0.01-1 parts of anti-oxidative stress component, 0.05-3 parts of soothing and repairing component, and an aqueous carrier, wherein the aqueous carrier is made up to 100 parts. The osmotic pressure protection component includes ectoine, the anti-oxidative stress component includes ergothioneine, and the soothing and repairing component includes panthenol and allantoin. Ectoine, ergothioneine, panthenol, and allantoin are all dispersed in the aqueous carrier.

[0024] The bilayer unpacking polydeoxyribonucleotide complex particle comprises a ternary core, an enzyme-sensitive shell, and a discontinuous lipid interlayer. The ternary core is formed by electrostatic recombination of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide; the enzyme-sensitive shell comprises low-molecular-weight sodium hyaluronate and trehalose, with the low-molecular-weight sodium hyaluronate located outside the ternary core and trehalose dispersed within the shell formed by the low-molecular-weight sodium hyaluronate; the discontinuous lipid interlayer comprises hydrogenated lecithin, ceramide NP, and cholesterol, and is composed of multiple spaced-apart sheet lipid regions distributed outside the enzyme-sensitive shell, with at least a portion of the sheet lipid regions embedded within the enzyme-sensitive shell.

[0025] The polydeoxyribonucleotides used in this application are purified sodium polydeoxyribonucleotides or their cosmetic-grade water-soluble form, with fragment lengths ranging from 80 to 600 bp. Fragment length can be confirmed by agarose gel electrophoresis, capillary electrophoresis, or high-performance liquid chromatography combined with standards. When the polydeoxyribonucleotide fragment length is less than 80 bp, the chain is too short, easily leading to a higher proportion of unencapsulated polydeoxyribonucleotides in the aqueous carrier; when the fragment length is greater than 600 bp, the system viscosity increases, and particle size enlargement and aggregation are more likely to occur during the formation of the ternary nucleus.

[0026] The quaternized chitosan oligosaccharides used in this application are selected from hydroxypropyltrimethylammonium chloride chitosan, N-trimethyl chitosan oligosaccharides, or other chitosan oligosaccharide derivatives with quaternary ammonium cationic groups, with a weight-average molecular weight of 1-10 kDa. The weight-average molecular weight can be determined by gel permeation chromatography. Quaternized chitosan oligosaccharides provide cationic sites, enabling them to form electrostatic complexes with negatively charged polydeoxyribonucleotides and, together with carnosine, constitute a ternary nucleus. When the weight-average molecular weight of the quaternized chitosan oligosaccharide is below 1 kDa, its ability to immobilize polydeoxyribonucleotides is insufficient; when the weight-average molecular weight is above 10 kDa, the ternary nucleus easily forms large-particle aggregates.

[0027] The low molecular weight sodium hyaluronate in this application has a weight-average molecular weight of 20-150 kDa, which can be determined by gel permeation chromatography. As the main film-forming component of the enzyme-sensitive shell, the low molecular weight sodium hyaluronate is located outside the ternary core. Trehalose is dispersed within the shell formed by the low molecular weight sodium hyaluronate. When the weight-average molecular weight of the low molecular weight sodium hyaluronate is below 20 kDa, the shell continuity is insufficient; when the weight-average molecular weight is above 150 kDa, the viscosity of the dispersion system increases, and the particle size of the composite particles tends to increase.

[0028] The discontinuous lipid interlayer is not a continuous liposome membrane that completely encapsulates the ternary core, but rather consists of multiple spaced-apart sheet-like lipid regions. Here, "intercalation" refers to a portion of the boundary of the sheet-like lipid region being located within the enzyme-sensitive shell region formed by low-molecular-weight sodium hyaluronate, rather than the sheet-like lipid region being completely free in the aqueous carrier.

[0029] The formation of discontinuous lipid interlayers was confirmed by transmission electron microscopy (TEM) or cryo-TEM. In TEM images, the continuous region surrounding the ternary nucleus was designated as the enzyme-sensitive shell region, and the lamellar structures distributed outside the enzyme-sensitive shell were designated as lamellar lipid regions. When at least a portion of the boundary of a lamellar lipid region lies within the enzyme-sensitive shell region, and the lamellar lipid region does not form a completely closed capsule, it is considered that the lamellar lipid region is embedded in the enzyme-sensitive shell. At least 30 particles were observed; if at least 60% of the particles had at least one lamellar lipid region embedded in the enzyme-sensitive shell, the discontinuous lipid interlayer was considered to have formed.

[0030] The average sheet thickness of the lamellar lipid regions was measured using transmission electron microscopy (TEM) or cryo-TEM. During measurement, at least 30 lamellar lipid regions were randomly selected, and the vertical distance between the two boundaries of each region was measured. The arithmetic mean of these distances was taken as the average sheet thickness. The average sheet thickness ranged from 5 to 40 nm. When the average sheet thickness was less than 5 nm, the boundaries of the lamellar lipid regions were not easily and stably identified; when the average sheet thickness was greater than 40 nm, lipid components were more likely to form free lipid aggregates or intact liposome structures.

[0031] The preparation method is as follows: First, prepare the ternary nucleus. Add polydeoxyribonucleotides to an aqueous buffer at pH 5.0-6.5 and stir until completely dissolved to obtain a polydeoxyribonucleotide solution. The aqueous buffer is an acetate buffer, citrate buffer, or phosphate buffer with a concentration of 5-50 mmol / L. Add carnosine to the polydeoxyribonucleotide solution and stir for 10-30 minutes to ensure uniform dispersion of the carnosine. Separately, dissolve quaternized chitosan oligosaccharide in an aqueous buffer at pH 5.0-6.5 to obtain a quaternized chitosan oligosaccharide solution.

[0032] At 20-30℃, a quaternized chitosan oligosaccharide solution is added dropwise to a mixed solution containing polydeoxyribonucleotides and carnosine, while stirring at 300-1000 rpm during the addition. After the addition is complete, stirring continues for 20-60 minutes, allowing the polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharides to form a ternary core through electrostatic recombination. In the ternary core, the mass ratio of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharides is 1:0.05-0.4:0.2-1.0.

[0033] Next, prepare the enzyme-sensitive shell. Add low-molecular-weight sodium hyaluronate to purified water and stir at 20-35℃ until completely dissolved to obtain a low-molecular-weight sodium hyaluronate solution. Add trehalose to the low-molecular-weight sodium hyaluronate solution and continue stirring until the trehalose is completely dissolved to obtain a low-molecular-weight sodium hyaluronate-trehalose solution. The mass ratio of low-molecular-weight sodium hyaluronate to trehalose is 1:0.3-2.5.

[0034] A low-molecular-weight sodium hyaluronate-trehalose solution was added to a ternary core dispersion and reacted for 20-60 minutes under stirring at 300-800 rpm. This resulted in the low-molecular-weight sodium hyaluronate being located on the outside of the ternary core, with trehalose dispersed within the shell formed by the low-molecular-weight sodium hyaluronate. The low-molecular-weight sodium hyaluronate-trehalose solution was added at a rate of 0.5-5 mL / min, and the pH of the reaction system was maintained at 5.0-6.5.

[0035] Then, a discontinuous lipid block was prepared. Hydrogenated lecithin, ceramide NP, and cholesterol were mixed at a mass ratio of 1:0.03-0.25:0.08-0.60, and added to ethanol, propylene glycol, or an aqueous phase containing ethanol. The mixture was stirred at 40-65°C to form a lipid pre-dispersion. The total lipid mass concentration in the lipid pre-dispersion was 0.1%-5%. If ethanol was used as a co-dispersing solvent, the residual ethanol was controlled within the allowable range for topical raw materials during subsequent vacuum removal or dilution steps.

[0036] The lipid pre-dispersion solution is added to the particle dispersion that has formed the enzyme-sensitive shell, and stirred at 35-55℃ for 10-40 minutes, followed by homogenization or ultrasonic treatment. The homogenization pressure is 20-80 MPa, and the cycle is 1-3 times; the ultrasonic treatment power is 100-500 W, and the treatment time is 1-10 minutes. Through the above treatment, hydrogenated lecithin, ceramide NP, and cholesterol form multiple spaced-apart sheet lipid regions, which are distributed on the outside of the enzyme-sensitive shell, and at least some of the sheet lipid regions are embedded in the enzyme-sensitive shell, resulting in bilayer unpacked polydeoxyribonucleotide complex particles.

[0037] With a total mass of 100 parts for the double-layer unpacking polydeoxyribonucleic acid complex particles, the ternary core consists of 20-55 parts, the enzyme-sensitive shell consists of 30-65 parts, and the discontinuous lipid interlayer consists of 5-20 parts, and the sum of the mass parts of the ternary core, the enzyme-sensitive shell, and the discontinuous lipid interlayer is 100 parts.

[0038] The mass fractions of the ternary core, enzyme-sensitive shell, and discontinuous lipid interlayer are calculated based on the actual bound mass of each component. The mass of the ternary core is the sum of the masses of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharides bound to the composite particles; the mass of the enzyme-sensitive shell is the sum of the masses of low-molecular-weight sodium hyaluronate and trehalose bound to the composite particles; and the mass of the discontinuous lipid interlayer is the sum of the masses of hydrogenated lecithin, ceramide NP, and cholesterol bound to the composite particles. Unbound components are separated from the composite particles by ultrafiltration centrifugation, dialysis, or centrifugal washing, and the content of the corresponding component in the separation solution is detected. The actual bound mass of each layer is the mass of the corresponding component fed into the composite particle minus the mass of the unbound component in the separation solution.

[0039] After obtaining the bilayer unpacked polydeoxyribonucleic acid (PDA) composite particles, they were mixed with an aqueous carrier. The aqueous carrier consisted of purified water, glycerol, and 1,3-propanediol, with the total mass of glycerol and 1,3-propanediol being 2%-12% of the aqueous carrier's mass. Ectoin, ergothioneine, panthenol, and allantoin were added to the aqueous carrier and stirred until uniformly dispersed. The mass ratio of panthenol to allantoin was 1:0.02-0.3. The bilayer unpacked PDA composite particles were then added, and the pH was adjusted to 5.0-6.5 to obtain a topical medical aesthetic active ingredient composition with repairing and anti-aging effects.

[0040] In this application, the polydeoxyribonucleotide loading rate is calculated according to the following formula: ; in, The loading rate of polydeoxyribonucleotides; The total mass of polydeoxyribonucleic acid added during the preparation of bilayer unpacked polydeoxyribonucleic acid complex particles; The mass of polydeoxyribonucleotides present in a continuous aqueous phase and not encapsulated in the ternary core after the preparation of composite particles. After separation by ultrafiltration centrifugation, dialysis, or gel size exclusion, the polydeoxyribonucleotides in the separated solution are determined by ultraviolet absorption, high-performance liquid chromatography, or fluorescence labeling. The polydeoxyribonucleotide loading rate is not less than 70%.

[0041] In this application, the polydeoxyribonucleotide release rate is calculated based on the cumulative release rate: ; ; in, for The cumulative release rate of polydeoxyribonucleotides at any given time; for The cumulative release mass of polydeoxyribonucleotides at any given time; The mass of polydeoxyribonucleic acid initially loaded onto the double-layer unpacking polydeoxyribonucleic acid complex particles; for The concentration of polydeoxyribonucleotides in the release medium is constantly monitored; To release the total volume of the medium; for Before the moment The concentration of polydeoxyribonucleic acid in the release medium during the second sampling; For the first Sampling volume; for Number of samples taken before the specified time; for The sum of the masses of polydeoxyribonucleotides carried out by each sample taken before the endpoint. When only one sample is taken at the endpoint during the release process, .

[0042] Release rate was determined using the dialysis bag method. The sample containing bilayer unpacked polydeoxyribonucleic acid (PDA) complex particles was placed in a dialysis bag, which was then placed in the release medium and shaken at 37°C. The molecular weight cutoff of the dialysis bag was selected to retain the bilayer unpacked PDA complex particles while allowing the released PDA fragments to pass through. The release medium was an aqueous system with a pH of 5.0-6.5, using acetate buffer, citrate buffer, or phosphate buffer. The volume of the release medium was 20-100 times the sample volume. After each sampling, an equal volume of fresh release medium was added, and correction was performed according to the cumulative release rate formula. At least three parallel analyses were performed on each sample group.

[0043] After being placed in an aqueous system at 37℃, pH 5.0-6.5, and without hyaluronidase for 4 hours, the polydeoxyribonucleotide release rate of the bilayer unpacking polydeoxyribonucleotide complex particles was no higher than 30%. After being placed in an aqueous system at 37℃, pH 5.0-6.5, and with a hyaluronidase concentration of 10-100 U / mL for 24 hours, the polydeoxyribonucleotide release rate of the bilayer unpacking polydeoxyribonucleotide complex particles was 55%-85%.

[0044] In this application, the particle size variation factor is calculated according to the following formula: ; in, This represents the multiple of particle size variation. The D50 particle size of the sample before hyaluronidase treatment; The D50 particle size of the sample is shown after t hours of hyaluronidase treatment. After 2 hours of incubation in an aqueous system at 37°C, pH 5.0-6.5, and a hyaluronidase concentration of 10-100 U / mL, the D50 particle size of the bilayer unpacked polydeoxyribonucleic acid (PDU) composite particles was 1.2-2.5 times that before incubation. Comparisons of D50 particle size before and after hyaluronidase treatment were performed using the same sample, the same dilution factor, and the same particle size analyzer parameters.

[0045] D50 particle size, polydispersity index, and zeta potential were determined using a nanoparticle size and zeta potential analyzer. During testing, samples were diluted to the instrument's recommended scattering intensity range using an aqueous medium at pH 5.5. The testing temperature was 25℃, and each sample was tested in parallel at least three times, with the average value taken. The D50 particle size of the bilayer unpacked polydeoxyribonucleotide composite particles ranged from 80 to 350 nm, the polydispersity index was no greater than 0.35, and the zeta potential ranged from -20 mV to +10 mV.

[0046] In the topical medical aesthetic active ingredient composition, the total content of polydeoxyribonucleotides is 0.005wt%-0.8wt%. Polydeoxyribonucleotides not encapsulated in the ternary nucleus refer to polydeoxyribonucleotides present in the external aqueous phase after separation by ultrafiltration, centrifugation, dialysis, or gel exclusion. The mass of polydeoxyribonucleotides not encapsulated in the ternary nucleus accounts for no more than 20% of the total mass of polydeoxyribonucleotides.

[0047] Example 1 Weigh out 0.10 parts of polydeoxyribonucleotide, 0.02 parts of carnosine, and 0.06 parts of quaternized chitosan oligosaccharide. Add the polydeoxyribonucleotide to an acetate buffer solution at pH 5.5 and stir until completely dissolved. Add the carnosine and continue stirring for 20 minutes. Separately, dissolve the quaternized chitosan oligosaccharide in an aqueous solution at pH 5.5 to obtain a quaternized chitosan oligosaccharide solution. Slowly add the quaternized chitosan oligosaccharide solution dropwise to the mixed solution of polydeoxyribonucleotide and carnosine, maintaining stirring at 500 rpm during the addition. After the addition is complete, continue stirring for 40 minutes to obtain a ternary core dispersion.

[0048] Weigh out 0.15 parts of low molecular weight sodium hyaluronate and 0.15 parts of trehalose. Dissolve the low molecular weight sodium hyaluronate in purified water by stirring, then add the trehalose and stir until dissolved to obtain a low molecular weight sodium hyaluronate-trehalose solution. Add the low molecular weight sodium hyaluronate-trehalose solution to the ternary core dispersion at a rate of 1 mL / min, maintaining the pH at 5.5, and continue stirring for 40 minutes. This allows the low molecular weight sodium hyaluronate to be located on the outside of the ternary core, while the trehalose is dispersed within the shell formed by the low molecular weight sodium hyaluronate, thus forming an enzyme-sensitive shell.

[0049] 0.05 parts of hydrogenated lecithin, 0.006 parts of ceramide NP, and 0.015 parts of cholesterol were weighed and added to a propylene glycol aqueous phase containing a small amount of ethanol. The mixture was stirred at 50°C to form a lipid pre-dispersion. The lipid pre-dispersion was added to a particle dispersion that had already formed an enzyme-sensitive shell. The mixture was stirred at 50°C for 20 minutes and then homogenized to obtain a bilayer unpacking polydeoxyribonucleotide composite particle dispersion. Transmission electron microscopy revealed multiple spaced-apart lamellar lipid regions on the outer side of the composite particles. The boundaries of some of these lamellar lipid regions were located within the enzyme-sensitive shell region, and the lamellar lipid regions did not form a completely closed capsule.

[0050] The above-mentioned double-layer unpacking polydeoxyribonucleotide composite particle dispersion was added to an aqueous carrier comprising purified water, glycerol, and 1,3-propanediol. 0.8 parts of ectoine, 0.05 parts of ergothioneine, 0.5 parts of panthenol, and 0.05 parts of allantoin were added to the aqueous carrier, the pH was adjusted to 5.5, and purified water was added to bring the total to 100 parts, resulting in a topical medical aesthetic active ingredient composition with repairing and anti-aging effects.

[0051] Example 2 The difference between this embodiment and Embodiment 1 is that the mass ratio of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide is adjusted to 1:0.1:0.4; the mass ratio of low molecular weight sodium hyaluronate and trehalose is adjusted to 1:1.0; and the mass ratio of hydrogenated lecithin, ceramide NP, and cholesterol is adjusted to 1:0.10:0.30. The remaining preparation steps are the same as in Embodiment 1. This embodiment is used to illustrate that, within the proportional range defined in the claims, by adjusting the ratio of the ternary core, the enzyme-sensitive shell, and the discontinuous lipid interlayer, bilayer unpacking polydeoxyribonucleotide composite particles with different particle sizes and release rates can be obtained.

[0052] Example 3 The difference between this embodiment and Embodiment 1 is that, after preparing the bilayer unpacked polydeoxyribonucleic acid (PDA) composite particle dispersion, trehalose and mannitol are added as freeze-drying protectants, followed by pre-freezing, primary drying, and secondary drying to obtain a freeze-dried topical medical aesthetic active ingredient. In use, the freeze-dried topical medical aesthetic active ingredient is reconstituted with purified water or an aqueous carrier containing glycerol and 1,3-propanediol, and ectoine, ergothioneine, panthenol, and allantoin are added to adjust the pH to 5.0-6.5. After reconstitution, the bilayer unpacked PDA composite particles are tested according to the aforementioned particle size, loading rate, and release rate detection methods to confirm whether they meet the requirements of a D50 particle size of 80-350 nm, a polydispersity index of no more than 0.35, a Zeta potential of -20 mV to +10 mV, and a PDA loading rate of no less than 70%.

[0053] To illustrate the difference between this application and ordinary compound systems and conventional coating systems, the following comparative examples can be set up.

[0054] Comparative Example 1 is a free mixture. Polydeoxyribonucleotides, carnosine, low molecular weight sodium hyaluronate, trehalose, hydrogenated lecithin, ceramide NP, and cholesterol were directly added to an aqueous carrier and stirred. No ternary core formation, enzyme-sensitive shell coating, or discontinuous lipid interlayer construction was performed. This comparative example was used to compare the proportion of polydeoxyribonucleotides not encapsulated in the ternary core with the examples.

[0055] Comparative Example 2 consists of particles without an enzyme-sensitive shell. A polydeoxyribonucleotide-carnosine-quaternized chitosan oligosaccharide ternary core was prepared according to the method of Example 1, but without the addition of low-molecular-weight sodium hyaluronate and trehalose to form an enzyme-sensitive shell, and without constructing a discontinuous lipid interlayer. This comparative example is used to compare the particle size change fold and polydeoxyribonucleotide release rate with the examples under the presence of hyaluronidase.

[0056] Comparative Example 3 consists of particles without discontinuous lipid interlayers. A ternary core and an enzyme-sensitive shell were prepared according to the method of Example 1, but hydrogenated lecithin, ceramide NP, and cholesterol were not added to form a discontinuous lipid interlayer. This comparative example was used to compare the early release rate under hyaluronidase-free conditions with the examples.

[0057] Comparative Example 4 is a physical mixture of ceramide liposomes. A ternary core and an enzyme-sensitive shell were prepared according to the method of Example 1. Liposomes formed from hydrogenated lecithin, ceramide NP, and cholesterol were then prepared separately, and the liposomes were physically mixed with the enzyme-sensitive shell particles. This comparative example illustrates that the physical mixing method of intact liposomes differs from the discontinuous lipid intercalation of this application. The liposomes are mainly free in the aqueous carrier and do not belong to a structure in which at least some sheet-like lipid regions are embedded in the enzyme-sensitive shell.

[0058] Comparative Example 5 shows PDRN particles coated with hyaluronic acid. Polydeoxyribonucleotides were formed into a complex with cationic materials, and then coated with hyaluronic acid. However, carnosine was not added to form a ternary core, trehalose was not dispersed in the enzyme-sensitive shell, and a discontinuous lipid interlayer was not incorporated. This comparative example illustrates the difference between ordinary PDRN hyaluronic acid-coated particles and the combined structure of the ternary core, low-molecular-weight sodium hyaluronate-trehalose enzyme-sensitive shell, and discontinuous lipid interlayer of this application.

[0059] The above embodiments and comparative examples were evaluated using the same methods for particle size detection, zeta potential detection, transmission electron microscopy observation, encapsulation rate detection, unencapsulated proportion detection, enzyme-free release detection, and hyaluronidase response release detection. Through comparison using the same detection methods, it can be confirmed that the topical medical aesthetic active ingredient composition of this application is not a common active ingredient blend, nor is it a physical mixture of PDRN hyaluronic acid coated particles, chitosan oligosaccharide hyaluronic acid aggregated particles, or ceramide liposomes. Instead, it is a layered composite particle system with a ternary core, an enzyme-sensitive shell, and a discontinuous lipid interlayer.

[0060] This application can evaluate topical repair performance using a keratinocyte scratch model, anti-aging performance using fibroblast collagen-related indicators, and anti-oxidative stress performance using a UVA-induced oxidative stress model. During evaluation, the sample from the examples, the free mixture, enzyme-sensitive shell particles, non-discontinuous lipid intercalation particles, and the ceramide liposome physical mixture were used as control groups. Repair performance was characterized by scratch closure rate; anti-aging performance was characterized by the expression levels of COL1A1, COL3A1, and MMP-1; and anti-oxidative stress performance was characterized by intracellular ROS levels. The above evaluations are used to illustrate the influence of the ternary core, enzyme-sensitive shell, and discontinuous lipid intercalation on the delivery structure of the topical active ingredient and the topical care performance. Before obtaining actual test results, efficacy conclusions are not recorded as specific percentages or multiples.

[0061] In this application, the ranges of each parameter are determined based on particle nucleation stability, shell coating integrity, external aqueous phase dispersion stability, and hyaluronidase response release characteristics. When the content of bilayer unpacking polydeoxyribonucleotide composite particles is less than 0.5 parts, the content of composite particles in the composition is too low, which is not conducive to the formation of a stable polydeoxyribonucleotide loading system; when it is higher than 8 parts, the viscosity of the aqueous phase system increases, and the particles are prone to collision and aggregation. When the ternary core is less than 20 parts, the polydeoxyribonucleotide loading is insufficient; when it is higher than 55 parts, the ratio of the outer enzyme-sensitive shell and the discontinuous lipid interlayer is insufficient. When the enzyme-sensitive shell is less than 30 parts, it cannot fully cover the ternary core; when it is higher than 65 parts, the particle size is prone to increase. When the discontinuous lipid interlayer is less than 5 parts, it is difficult to form observable sheet lipid regions; when it is higher than 20 parts, the sheet lipid regions are prone to transform into free liposomes or lipid aggregates.

[0062] In this application, the D50 particle size of 80-350 nm, polydispersity index of no more than 0.35, and zeta potential of -20 mV to +10 mV are defined to ensure dispersion stability in the external aqueous phase system. When the particle size is below 80 nm, the particle surface area is large, and the early release rate is easily increased; when the particle size is above 350 nm, the system dispersion uniformity decreases. When the polydispersity index is above 0.35, the particle size distribution is wide, resulting in insufficient batch stability. When the zeta potential exceeds the range of -20 mV to +10 mV, the system may exhibit excessively strong negative or positive charges, which is detrimental to the dispersion stability in external formulations.

[0063] In this application, after 4 hours of incubation in an aqueous system without hyaluronidase, the polydeoxyribonucleotide release rate was no higher than 30%, confirming that the ternary core, enzyme-sensitive shell, and discontinuous lipid interlayer could reduce early release. After 24 hours of incubation in an aqueous system containing hyaluronidase, the polydeoxyribonucleotide release rate was 55%-85%, confirming that the enzyme-sensitive shell was responsive to hyaluronidase. After 2 hours of incubation in an aqueous system containing hyaluronidase, the D50 particle size was 1.2-2.5 times the original D50 particle size, confirming that the enzyme-sensitive shell loosened and its hydrated volume increased under the action of hyaluronidase, rather than simply undergoing ordinary diffusion release.

[0064] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A medical and cosmetic active material composition having a repair anti-aging effect, characterized by, It includes 0.5-8 parts of double-layer unpacking polydeoxyribonucleic acid complex particles, 0.2-4 parts of osmotic pressure protection component, 0.01-1 parts of antioxidant stress component, 0.05-3 parts of soothing and repairing component, and an aqueous carrier, with the aqueous carrier supplemented to 100 parts; Osmotic protection components include ectoine, antioxidant stress components include ergothioneine, and soothing and repairing components include panthenol and allantoin. The double-layer unpacking polydeoxyribonucleic acid complex particle includes a ternary core, an enzyme-sensitive shell, and a discontinuous lipid interlayer; The ternary core is formed by electrostatic recombination of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide. The enzyme-sensitive shell consists of low-molecular-weight sodium hyaluronate and trehalose. The low-molecular-weight sodium hyaluronate is located outside the ternary core, and the trehalose is dispersed in the shell formed by the low-molecular-weight sodium hyaluronate. The discontinuous lipid block includes hydrogenated lecithin, ceramide NP and cholesterol. The discontinuous lipid block is composed of multiple sheet lipid regions spaced apart from each other. The sheet lipid regions are distributed on the outside of the enzyme-sensitive shell, and at least some of the sheet lipid regions are embedded in the enzyme-sensitive shell.

2. The medical aesthetic active ingredient composition having a repair anti-aging effect according to claim 1, wherein With a total mass of 100 parts for the double-layer unpacked polydeoxyribonucleotide complex particles, the ternary core comprises 20-55 parts, the enzyme-sensitive shell comprises 30-65 parts, and the discontinuous lipid interlayer comprises 5-20 parts, and the sum of the mass parts of the ternary core, the enzyme-sensitive shell, and the discontinuous lipid interlayer is 100 parts.

3. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 2, characterized in that, In the ternary core, the mass ratio of polydeoxyribonucleotides, carnosine, and quaternized chitosan oligosaccharide is 1:0.05-0.4:0.2-1.0; The length of the polydeoxyribonucleotide fragment is 80-600 bp, and the weight-average molecular weight of the quaternized chitosan oligosaccharide is 1-10 kDa.

4. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 3, characterized in that, In the enzyme-sensitive shell, the mass ratio of low molecular weight sodium hyaluronate to trehalose is 1:0.3-2.5; The weight-average molecular weight of the low molecular weight sodium hyaluronate is 20-150 kDa.

5. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 4, characterized in that, In the discontinuous lipid block, the mass ratio of hydrogenated lecithin, ceramide NP, and cholesterol is 1:0.03-0.25:0.08-0.60; The average sheet thickness of the lipid sheet region was measured to be 5-40 nm using transmission electron microscopy.

6. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 5, characterized in that, The double-layer unpacking polydeoxyribonucleotide composite particles have a D50 particle size of 80-350 nm, a polydispersity index of no more than 0.35, a zeta potential of -20 mV to +10 mV, and a polydeoxyribonucleotide loading rate of no less than 70%.

7. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 6, characterized in that, After being placed in an aqueous system at 37℃, pH 5.0-6.5, and without hyaluronidase for 4 hours, the release rate of polydeoxyribonucleic acid (PDA) from the bilayer unpacking PDA complex particles was no higher than 30%. After being placed in an aqueous system at 37℃, pH 5.0-6.5, and hyaluronidase concentration of 10-100 U / mL for 24 hours, the polydeoxyribonucleotide release rate of the double-layer unpacking polydeoxyribonucleotide composite particles was 55%-85%. The release rate is calculated as the ratio of the mass of polydeoxyribonucleic acid (PDA) in the release medium to the initial mass of PDA in the bilayer unpacked PDA composite particles.

8. The medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 7, characterized in that, After being placed in an aqueous system at 37℃, pH 5.0-6.5 and hyaluronidase concentration of 10-100 U / mL for 2 hours, the D50 particle size of the double-layer unpacked polydeoxyribonucleic acid complex particles was 1.2-2.5 times that before placement.

9. A medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 8, characterized in that, The mass ratio of panthenol to allantoin is 1:0.02-0.3; Ectoin, ergothioneine, panthenol, and allantoin are all dispersed in an aqueous carrier.

10. A medical aesthetic active ingredient composition with repairing and anti-aging effects according to claim 9, characterized in that, The aqueous support comprises purified water, glycerol, and 1,3-propanediol, wherein the total mass of glycerol and 1,3-propanediol is 2%-12% of the mass of the aqueous support. The pH of the medical aesthetic active ingredient composition is 5.0-6.5, the total content of polydeoxyribonucleotides in the medical aesthetic active ingredient composition is 0.005wt%-0.8wt%, and the mass of polydeoxyribonucleotides not encapsulated in the ternary core accounts for no more than 20% of the total mass of polydeoxyribonucleotides.

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