Compositions for repairing mitochondrial mtDNA damage and anti-aging, and preparation method and application thereof

Abstract

The application discloses a composition for repairing mitochondrial mtDNA damage and anti-aging, a preparation method and application thereof, and belongs to the technical field of cosmetics or similar toiletry preparations. According to weight parts, the composition comprises 0.01-0.5 parts of acrylic acid (ester) / C10-30 alkyl alcohol acrylic acid ester cross-linked polymer, 0.05-10.0 parts of microspheric PDRN, 0.01-10.0 parts of mitochondrial PDRN, 0.01-2 parts of inositol, 0.01-2.5 parts of a skin feel agent, 1-30 parts of a humectant and 0.001-0.2 parts of an anti-inflammatory soothing ingredient, and the rest is water. The composition is formed by double PDRN targeted delivery, mitochondrial metabolism empowerment, anti-inflammatory soothing and multi-dimensional component compounding of barrier moisturizing, and forms systematic synergistic gain from three dimensions of delivery stability, cell repair pathway and energy metabolism regulation, and synergistically realizes the effects of mitochondrial mtDNA damage repair and skin anti-aging, and significantly improves the anti-aging repair effect.

Description

Technical Field

[0001] This invention belongs to the technical field of cosmetics or similar grooming products, specifically relating to compositions for repairing mitochondrial mtDNA damage and anti-aging, their preparation methods, and applications. Background Technology

[0002] Current mitochondrial anti-aging skincare technologies primarily rely on PDRN (polydeoxyribonucleotide, INCI name sodium DNA), an ingredient mainly derived from salmon germ cell extracts. Its base composition shares 98% similarity with human DNA, leading to its widespread use. Existing technologies mainly enhance its transdermal absorption rate through microsphere formation, supramolecular encapsulation, or genetic engineering synthesis, and it is often combined with ingredients such as carnosine, yeast fermentation products, and sodium hyaluronate to achieve multiple effects including anti-aging, moisturizing, and barrier repair. In addition, NAD+ enhancers (such as NMN), antioxidants (such as ergothioneine), and peptide delivery technologies are also important pathways for mitochondrial protection.

[0003] However, existing mitochondrial anti-aging skincare technologies have the following problems: First, low transdermal absorption efficiency. PDRN is a large-molecule nucleic acid substance with a molecular weight range of 50-1500 KD, far exceeding the threshold of small molecules that the stratum corneum of the skin can freely penetrate. The stratum corneum of human skin has a natural barrier function. Under conventional topical application methods, most PDRN can only remain on the skin surface and cannot penetrate the stratum corneum to reach the dermis and the mitochondrial action sites of skin cells, making it difficult to exert endogenous repair and anti-aging effects. Currently, improved delivery technologies such as microsphereization can only slightly improve penetration efficiency, with limited improvement effects. To ensure efficacy, most highly active PDRN products still rely on medical aesthetic injections to deliver the drug through skin penetration. Home-based, non-invasive topical skincare products have always been unable to overcome the core bottleneck of transdermal absorption, greatly limiting the use of this type of ingredient. Second, insufficient synergy in the efficacy of the formulation system, and the problem of fragmented action pathways in multi-ingredient combinations. Current PDRN compound formulations are mostly simple physical mixtures, relying solely on the synergistic effects of each ingredient's individual properties. For example, PDRN focuses on basic DNA and mitochondrial repair, while NAD+ enhancers focus on cellular energy replenishment. The targets and pathways of action of various active ingredients are independent, failing to effectively improve skin aging problems. Furthermore, the simple combination of active ingredients with different physicochemical properties easily leads to issues such as ingredient antagonism and decreased system stability, further reducing the overall formula's actual anti-aging and repair effects. In summary, current mitochondrial anti-aging products suffer from low transdermal absorption efficiency and insufficient synergistic effects. The industry urgently needs a mitochondrial anti-aging solution that can efficiently improve PDRN transdermal absorption while achieving synergistic effects across the entire ingredient pathway. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the present invention aims to provide a composition for repairing mitochondrial mtDNA damage and anti-aging, as well as its preparation method and application, to solve the technical problems of low transdermal absorption efficiency and insufficient synergistic efficacy of existing anti-aging products.

[0005] To achieve the above objectives, the present invention employs the following technical solution: The first aspect of the present invention discloses a composition for repairing mitochondrial mtDNA damage and anti-aging, comprising, by weight: 0.01 to 0.5 parts of acrylate / C10-30 alkanol acrylate crosspolymer, 0.05 to 10.0 parts of microsphere-derived PDRN, 0.01 to 10.0 parts of mitochondrial PDRN, 0.01 to 2 parts of inositol, 0.01 to 2.5 parts of a skin feeler, 1 to 30 parts of a moisturizer, and 0.001 to 0.2 parts of an anti-inflammatory and soothing ingredient, with the balance being water.

[0006] Preferably, it also includes 0.001 to 0.2 parts of urolithiasis A.

[0007] Preferably, the skin-feeling agent is mannitol and / or dextran.

[0008] Preferably, the humectant is any one or more of butylene glycol, glycerin, and 1,2-pentanediol.

[0009] Preferably, the anti-inflammatory and soothing ingredients are sodium heparin and / or asiaticoside.

[0010] Preferably, the microsphere-shaped PDRN is a polyglycoside. ® Microsphere-modified PDRN, wherein the mitochondrial PDRN is polyglycoside. ® Mitochondrial PDRN.

[0011] Preferably, by weight, it comprises: 0.01 to 0.5 parts of acrylate / C10-30 alkanol acrylate crosspolymer, and 0.05 to 10.0 parts of polyglycoside. ® Microsphere-sized PDRN, 0.01~10.0 parts of polyglycoside ® Mitochondrial PDRN, 0.01-2 parts inositol, 0.01-2.5 parts mannitol, 0.2-7 parts butylene glycol, 0.2-7 parts glycerol, 0.6-16 parts 1,2-pentanediol and 0.001-0.2 parts sodium heparin, with the balance being water.

[0012] In a second aspect, the present invention discloses a method for preparing the above-mentioned composition for repairing mitochondrial mtDNA damage and anti-aging. An acrylate / C10-30 alkanol acrylate crosspolymer and a portion of a moisturizer are dissolved in water at 70-85°C. A skin-feeling agent and anti-inflammatory soothing ingredients are added and mixed. The mixture is then cooled to 30-55°C, and microsphere-type PDRN, mitochondrial PDRN, inositol, the remaining moisturizer, and water are added and mixed to obtain the composition for repairing mitochondrial mtDNA damage and anti-aging.

[0013] A third aspect of the present invention discloses the application of the above-described composition for repairing mitochondrial mtDNA damage and anti-aging in the preparation of anti-aging products.

[0014] A fourth aspect of the present invention discloses a mitochondrial damage repair agent containing the above-mentioned composition for repairing mitochondrial mtDNA damage and anti-aging.

[0015] Compared with the prior art, the present invention has the following beneficial effects: The composition for repairing mitochondrial mtDNA damage and anti-aging provided by this invention comprises: 1) an acrylate / C10-30 alkyl acrylate crosspolymer that can form a three-dimensional network structure, regulate the rheological properties of the system, significantly increase the viscosity of the system, ensure a rich feel when applied, and quickly spread to form a thin and breathable film on the skin surface, avoiding a heavy and sticky feeling and improving the user experience; combined with a moisturizer, it forms a water-locking protective film on the skin surface, reducing skin moisture loss and alleviating skin dryness and barrier fragility problems associated with mitochondrial damage, repairing mitochondria from the inside and strengthening the skin barrier from the outside, thus synergistically enhancing the skin's anti-aging ability; 2) microsphere-based PDRN, relying on the microsphere carrier structure, can penetrate the skin's stratum corneum barrier and carry nucleic acid active ingredients. Microsphere PDRN penetrates the epidermis to reach the dermis, overcoming the limitations of conventional large-molecule PDRNs that cannot penetrate the skin and remain only on the surface. Mitochondrial PDRNs, with their mitochondrial targeting capabilities, can precisely penetrate cell and mitochondrial membranes, directly acting on the mitochondria within skin cells. They specifically repair core issues such as mitochondrial mtDNA breaks and base damage caused by UV radiation and oxidative stress, blocking mitochondrial functional decline at its intrinsic cellular level and significantly improving the bioavailability of nucleic acid active ingredients. Photodamage repair experiments further demonstrate that microsphere PDRNs and mitochondrial PDRNs can synergistically reduce MDA levels and increase NAD+ levels. +The ratio of NADH and mtDNA copy number has the effect of repairing mitochondrial mtDNA damage and anti-wrinkle and firming effects; 3) The formula is compounded with inositol, which can effectively regulate mitochondrial membrane potential, improve mitochondrial ATP energy synthesis efficiency, clear reactive oxygen free radicals accumulated in mitochondria, and reduce secondary damage to mtDNA caused by oxidative stress; at the same time, inositol can regulate cellular lipid metabolism, assist in the repair of damaged cell membrane structure, and form a closed-loop anti-aging pathway of "free radical scavenging-mitochondrial energy replenishment-mtDNA damage repair" with dual PDRN, which improves the problem of premature mitochondrial aging in all dimensions; 4) By adding anti-inflammatory and soothing ingredients, it can inhibit the chronic micro-inflammation of the skin accompanied by mitochondrial damage, reduce the skin discomfort caused by active ingredients, and further reduce the continuous attack of inflammatory factors on mtDNA after the inflammation is relieved, block the vicious cycle of inflammation-mitochondrial damage-skin aging, and broaden the range of skin types suitable for the composition; 5) The addition of skin feeler can improve the stickiness defect caused by macromolecular nucleic acid raw materials, and improve the smoothness of application and the speed of skin absorption. Therefore, this composition, through the combination of multiple components including dual PDRN targeted delivery, mitochondrial metabolism empowerment, anti-inflammatory soothing, and barrier moisturizing, forms a systematic synergistic gain from three dimensions: delivery stability, cell repair pathway, and energy metabolism regulation. It synergistically achieves mitochondrial mtDNA damage repair and skin anti-aging effects, significantly improving the anti-aging and repair effect.

[0016] Furthermore, the added urolithin A in the composition can promote skin penetration and cellular absorption of microsphere-based PDRN and mitochondrial-targeted PDRN, and stabilize mitochondrial structure. Experiments using MDA, NAD+ / NADH ratios, and mtDNA copy number assays show that within cells, urolithin A activates mitophagy to specifically clear impaired mitochondria. Simultaneously, mitochondrial PDRN is delivered to the mitochondria, providing nucleotide raw materials for damaged mtDNA and directly promoting its repair. Meanwhile, microsphere-based PDRN plays a broad role in DNA repair and cell regeneration promotion in the nucleus and cytoplasm. These three elements work synergistically to form a complete anti-aging closed loop of "damage clearance - precise repair - comprehensive regeneration." In addition, urolithin A, while clearing old mitochondria, can induce mitochondrial biogenesis, increasing the "quantity" and "quality" of the cell's overall energy factories. The repaired, functional mitochondria (thanks to PDRN repair) can produce ATP more efficiently. Degradation capacity testing experiments demonstrated that urolithin A, through its polyphenolic structure, forms intermolecular forces with the carrier of microsphere-derived PDRN, increasing the structural integrity of the microspheres in the physiological environment from 60% to 88%. This significantly prolongs the sustained release and action time of the active ingredient, solving the problem of sodium DNA degradation due to light, oxidation, temperature, and enzymatic hydrolysis, and improving the stability of the composition. Through the specific combination of urolithin A, microsphere-derived PDRN, and mitochondrial PDRN, multiple synergies are achieved in three dimensions: physical delivery stability, cellular pathways, and energy metabolism. Ultimately, this results in superior mitochondrial mtDNA repair and skin anti-aging effects that cannot be achieved by any single component and are not simply additive.

[0017] The preparation method of the composition for repairing mitochondrial mtDNA damage and anti-aging provided by the present invention is simple, highly operable, and easy to implement. The resulting composition for repairing mitochondrial mtDNA damage and anti-aging has a stable formulation, is suitable for sensitive skin, and can quickly repair damaged skin. Attached Figure Description

[0018] Figure 1 The graph shows the MDA (nmol / mg) test results of the compositions in different experimental groups of this invention; Figure 2 This is a graph showing the NAD+ / NADH ratio of the compositions in different experimental groups of this invention. Figure 3 This is a summary graph of the mtDNA copy number results of different experimental groups of the present invention; Figure 4 The graph shows the degradation ability test results of the composition for repairing mitochondrial mtDNA damage and anti-aging according to the present invention. Detailed Implementation

[0019] To enable those skilled in the art to understand the features and effects of the present invention, the following description and definitions are only general descriptions of the terms and expressions mentioned in the specification. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in case of conflict, the definitions in this specification shall prevail.

[0020] The theories or mechanisms described and disclosed herein, whether right or wrong, should not in any way limit the scope of the invention, that is, the contents of the invention can be implemented without being limited by any particular theory or mechanism.

[0021] In this document, all features defined by numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are for the sake of brevity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered as covering and specifically disclosing all possible sub-ranges and individual numerical values ​​(including integers and fractions) within those ranges.

[0022] In this article, unless otherwise specified, “contains,” “includes,” “containing,” “has,” or similar terms cover the meanings of “composed of” and “mainly composed of,” for example, “A contains a” covers the meanings of “A contains a and others” and “A contains only a.”

[0023] For the sake of brevity, not all possible combinations of the technical features in each implementation scheme or embodiment are described herein. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each implementation scheme or embodiment can be combined arbitrarily, and all possible combinations should be considered within the scope of this specification.

[0024] This invention provides a composition for repairing mitochondrial mtDNA damage and anti-aging, comprising, by weight: 0.01-0.5 parts of acrylate / C10-30 alkanol acrylate crosspolymer, 0.05-10.0 parts of microsphere-type PDRN, 0.01-10.0 parts of mitochondrial PDRN, 0-0.2 parts of urolithin A, 0.01-2 parts of inositol, 0.01-2.5 parts of skin feeler, 1-30 parts of moisturizer, and 0.001-0.2 parts of anti-inflammatory and soothing ingredients, with the balance being water; Preferably, 0.3 parts of the acrylate / C10-30 alkanol acrylate crosslinking polymer are used. The microsphere-shaped PDRN is preferably polyglycoside. ® Microsphere-sized PDRN, preferably at a content of 1.5 parts; the mitochondrial PDRN is preferably polyglycoside phthalate. ®The mitochondrial PDRN content is preferably 2.5 parts; the skin feeler is, but is not limited to, mannitol and / or dextran, with a content preferably 0.1 parts; the moisturizer is any one or more of butylene glycol, glycerin and 1,2-pentanediol, with a content preferably 8 parts; the anti-inflammatory and soothing ingredients are, but are not limited to, sodium heparin and / or asiaticoside, with a content preferably 0.001 parts; the water is preferably purified water.

[0025] In the above-mentioned composition for repairing mitochondrial mtDNA damage and anti-aging, the functions of each component are as follows: Acrylic (ester) / C10-30 alkanol acrylate crosslinking polymer: As a carbomer polymer, it significantly increases the viscosity of the system by swelling in the aqueous phase to form a three-dimensional network structure, enabling the product to achieve a suitable consistency. This crosslinking polymer can create a unique "moisturizing" skin feel—it has a certain consistency to ensure a rich feel when applied, while also spreading quickly and forming a thin, breathable film on the skin surface, avoiding a heavy, sticky feeling and improving the user experience. Manufacturer: Lubrizol.

[0026] Polyglycoside ® Microsphere-sized PDRN: Polygluconate ® Microsphere technology uses polysaccharides or derived targeted polysaccharides as precursors. Leveraging or artificially designing weak interactions between molecules in each layer, and by loading peptides, proteins, nucleic acid molecules, or other active small molecules, it enables the layers to spontaneously associate, forming structurally complete, stable molecular aggregates or supramolecular structures with specific functions. Polydeoxyribonucleotides (PDRNs) are polynucleotides with a molecular weight range of 50 to 1500 kDa, containing 50% double-stranded deoxyribonucleotides and 90% single-stranded deoxyribonucleotides. Microsphere technology results in superior transdermal absorption and targeted recognition capabilities, repairing damaged skin cells and maintaining long-term skin stability. (Polyglycosides) ® The preparation process of microsphere PDRN is based on Chinese Patent Publication No. CN 117017824 B, manufactured by Baihong Synthetic Biotechnology (Yantai) Co., Ltd., with a particle size of 121 nm.

[0027] Polyglycoside ®Mitochondrial PDRN: Its molecular structure is highly similar to human mitochondrial DNA, enabling it to specifically recognize and integrate into damaged mitochondrial mtDNA. It activates the DNA repair enzyme system, directly repairing mitochondrial DNA breaks and mutations caused by oxidative stress, ultraviolet radiation, etc., thus addressing the functional decline of the cell's energy factories at its root. By activating the adenosine A2A receptor and AKT signaling pathway, it promotes mitochondrial ATP synthesis, enhances cellular energy levels, improves mitochondrial morphology and function, and reverses energy metabolism disorders related to cellular aging. Manufacturer: Baihong Synthetic Biotechnology (Yantai) Co., Ltd., particle size 184 nm.

[0028] Inositol: By inhibiting the AMPK-DRP1 pathway, it reduces excessive mitochondrial division, maintains cristae structure integrity, increases ATP production, blocks RO rebound, generates phospholipid derivatives such as IP3 and PIP2, and rapidly releases intracellular calcium. 2+ It activates calcium-dependent repair enzymes (such as transglutaminase), making PDRN-induced collagen cross-linking stronger. As the head group of phosphatidylinositol, it stabilizes the lipid bilayer, reduces transepidermal water loss by 12%, and helps hydrophilic adenosine and PDRN cross the membrane, increasing bioavailability by approximately 1.4 times. Manufacturer: Finnfeeds Finland Oy.

[0029] Urolithin A: Molecular formula is C 13 H8O4, chemically named 3,8-dihydroxybenzo[c]chromen-6-one, activates mitophagy, clears damaged mitochondria, and promotes cellular energy metabolism and regeneration. Manufacturer: Xi'an Kangwei Biotechnology.

[0030] This invention provides a method for preparing a composition for repairing mitochondrial mtDNA damage and anti-aging. 0.01-0.5 parts of an acrylate / C10-30 alkanol acrylate crosspolymer and a portion of a moisturizer are dissolved in water at 70-85°C and stirred until homogeneous to obtain phase A. 0.01-2.5 parts of a skin-feeling agent and 0.001-0.2 parts of an anti-inflammatory and soothing ingredient are added to phase A and mixed thoroughly. The mixture is then cooled to 30-55°C. Next, 0.05-10.0 parts of microsphere-derived PDRN, 0.01-10.0 parts of mitochondrial PDRN, 0-0.2 parts of urolithin A, 0.01-2 parts of inositol, the remaining moisturizer, and water are added and mixed thoroughly. The mixture is stirred for 15-30 minutes to obtain the composition for repairing mitochondrial mtDNA damage and anti-aging.

[0031] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading this description, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined in this application.

[0032] The following examples use instruments and equipment conventional in the art. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. All raw materials used in the following examples are conventional commercially available products with specifications conventional in the art. In this specification and the following examples, unless otherwise specified, "%" represents weight percentage, "parts" represents parts by weight, and "ratio" represents weight proportion.

[0033] I. Preparation of a composition for repairing mitochondrial mtDNA damage and anti-aging Table 1. Formulations of Examples and Comparative Examples

[0034] Example 1 The preparation method of the composition for repairing mitochondrial mtDNA damage and anti-aging is as follows: Step 1: Add 0.3 parts of acrylate / C10-30 alkanol acrylate crosspolymer and 4 parts of butanediol to purified water, and stir evenly at 85°C to obtain phase A; add 1.5 parts of polyglycidol... ® Microsphere-sized PDRN and 2.5 parts polyglycoside ® Mitochondrial PDRN was added to purified water and mixed and stirred for 30 min to obtain phase B. Step 2: Add 0.1 parts mannitol and 0.001 parts heparin sodium to phase A obtained in step 1, stir well, then cool to 45°C, and then add phase B, 0.1 parts inositol, 2 parts glycerol and 2 parts 1,2-pentanediol to the system, stir well, and obtain a composition for repairing mitochondrial mtDNA damage and anti-aging.

[0035] Example 2 The difference from Example 1 is the amount of the ingredients used. Referring to Table 1 and the preparation method of Example 1, a composition for repairing mitochondrial mtDNA damage and anti-aging was obtained.

[0036] Comparative Example 1 The difference from Example 1 is that the amounts of the components are different. Referring to Table 1 and the preparation method of Example 1, control composition 1 was obtained.

[0037] Comparative Example 2 The difference from Example 1 is the amount of the components used. Referring to Table 1 and the preparation method of Example 1, control composition 2 was obtained.

[0038] Comparative Example 3 The difference from Example 1 is the amount of the components used. Referring to Table 1 and the preparation method of Example 1, control composition 3 was obtained.

[0039] Comparative Example 4 The difference from Example 1 is that the amounts of the components are different. Referring to Table 1 and the preparation method of Example 1, control composition 4 was obtained.

[0040] II. Efficacy Testing of Compositions for Repairing Mitochondrial mtDNA Damage and Anti-Aging Photoaging of the skin is an important component of skin aging, with UVB (ultraviolet light) in the wavelength range of 290-320 nm being the main contributing factor. In a fibroblast photoaging model, repeated UVB irradiation leads to an imbalance in the redox system of fibroblasts, specifically the excessive production of reactive oxygen species (ROS), which consume large amounts of intracellular reducing substances, ultimately resulting in cumulative oxidative damage and accelerated aging. Malondialdehyde (MDA) is one of the end products of free radical-induced peroxidation of unsaturated lipids, forming lipid peroxides. It can combine with proteins, nucleic acids, or lipids to form insoluble substances, hardening biological membranes, reducing permeability, affecting cellular material exchange, and ultimately causing cell rupture and death. Measuring MDA levels can detect the level of lipid oxidation; therefore, MDA is used as an indicator of lipid oxidation. Coenzyme I (NAD) and reduced coenzyme I (NADH) participate in numerous metabolic reactions. The NAD* / NADH ratio is a key parameter reflecting cellular metabolism and redox status. Its imbalance leads to cellular metabolic disorders. Elevated ROS levels lead to oxidative damage to mitochondrial DNA (mtDNA), increasing the mutation rate and causing mitochondrial functional decline, resulting in signs of skin aging such as wrinkles and discoloration. This test evaluates the effectiveness of repairing mitochondrial mtDNA damage based on the detection of mtDNA, NAD*NADH, and MDA levels induced by UVB-induced photodamage in skin fibroblasts.

[0041] 1. Detection of MDA based on UVB-induced photodamage to skin fibroblasts The compositions prepared in Comparative Examples 1-4 and Examples 1 and 2 were used for testing. HFB cells at passage number P6 were revived and seeded into culture flasks, cultured at 37°C in a 5% CO2 incubator, and passaged when the cells reached 70%-80% confluence. The digested cells were resuspended in culture medium and counted for later use. Cell seeding volume 1 10 5Cells were seeded into 6-well plates at 37°C and incubated overnight in a microplate incubator (5% CO2), with a seeding volume of 2 mL per well. The drug dosage (comparative examples 1-4, and the compositions prepared in Examples 1 and 2) was 2 mL per well. Cells were irradiated with 700 mJ of UVB, with a negative control group (no sample added after UVB induction). After 48 h of cell incubation, the supernatant was discarded, and cells were washed with 1 mL of PBS in each well and collected. Cells were processed according to the MDA assay kit instructions, and finally, the MDA content of each group was measured at 532 nm using a microplate reader.

[0042] The results are as follows Figure 1 As shown, the MDA content in the negative control group and comparative examples 1-4 did not differ significantly, with contents of 4, 3.99, 3.89, 3.78, and 3.7, respectively. In contrast, the MDA content in the compositions of Examples 1 and 2 was 2.43 and 2.11, respectively, indicating that the content of polyglycosides... ® Microsphere-shaped PDRN, Polyglycan ® Mitochondrial PDRN can reduce MDA levels, and urolithin A can promote polyglycoside A. ® Microsphere-shaped PDRN, Polyglycan ® The mitochondrial PDRN composition effectively reduces MDA levels and has the effect of repairing mitochondrial mtDNA damage.

[0043] 2. Detection of NAD+ / NADH based on UVB-mediated photodamage to skin fibroblasts HFB cells with a substitution number of P6 were taken from the liquid ammonia tank, revived, and seeded into culture flasks. The cells were cultured at 37°C in a 5% CO2 incubator. When the cells reached 70%–80% confluence, they were digested and passaged. The digested cells were resuspended in culture medium and counted for later use. Cell seeding was carried out at a rate of 1... 10 5 Cells were seeded at a rate of 2 cells / well into 6-well plates and incubated overnight in an incubator (37°C, 5% CO2). Each well contained 2 mL of the drug (comparative examples 1-4, and the compositions prepared in Examples 1 and 2). Cells were irradiated with 700 mJ of UVB, with a negative control group (no sample added after UVB induction). After 48 h of cell incubation, the supernatant was discarded, and each well was washed with 1 mL of PBS according to NAD... + The NADH assay kit (WST-8 assay) instructions describe how to process cells, use a microplate reader for detection, and calculate NAD+ and NADH levels based on a standard curve, and then calculate the NAD+ content. + / NADH ratio.

[0044] The results are as follows Figure 2As shown, the NAD+ / NADH ratios in the negative control group and Comparative Examples 1-4 were not significantly different, with contents of 27.66, 28.44, 28.33, 28.45, and 28.43, respectively. In contrast, the NAD+ / NADH ratios in Examples 1 and 2 were 33.12 and 35.75, respectively, indicating that the NAD+ / NADH ratios in Polygluconate were significantly higher. ® Microsphere-shaped PDRN, Polyglycan ® Mitochondrial PDRN can increase NAD + The ratio of NADH, and urolithiasis A can promote polyglycoside A ® Microsphere-shaped PDRN, Polyglycan ® Mitochondrial PDRN composition better enhances NAD + The ratio of NADH has the effect of repairing mitochondrial mtDNA damage.

[0045] 3. mtDNA detection based on UVB-induced photodamage to skin fibroblasts Comparative Examples 1-4 and Examples 1 and 2 were subjected to mtDNA detection based on UVB-induced photodamage in skin fibroblasts. For cell resuscitation and culture, HFB cells of passage P6 were taken from a liquid nitrogen tank, resuscitated, and seeded into culture flasks. They were cultured at 37°C in a 5% CO2 incubator. When the cells reached 70%-80% confluence, they were digested and passaged. The digested cells were resuspended in culture medium and counted for later use. (The last sentence appears to be incomplete and possibly refers to a separate process.) 10 5 Cells were seeded into 6-well plates at a seeding rate of 1 cell per well and incubated overnight in an incubator (37°C, 5% CO2). The dosage per well (comparative examples 1-4, and the compositions prepared in Examples 1 and 2) was 2 mL. The cells were irradiated with 700 mJ of UVB. A negative control group without any sample was used after UVB induction. DNA extraction and RT-PCR were performed, and the mtDNA copy number was calculated.

[0046] The results are as follows Figure 3 As shown, the mtDNA copy numbers of the negative control group and comparative examples 1-4 were not significantly different, with contents of 164, 164, 165, 166, and 166, respectively. In contrast, the mtDNA copy numbers of Examples 1 and 2 were 184 and 195, respectively, indicating that the mtDNA copy numbers of Polyglucopyranoside... ® Microsphere-shaped PDRN, Polyglycan ® Mitochondrial PDRN can increase mtDNA copy number, and urolithin A can promote polyglycoside A. ® Microsphere-shaped PDRN, Polyglycan ® The mitochondrial PDRN composition better increases mtDNA copy number and has the effect of repairing mitochondrial mtDNA damage.

[0047] 4. Anti-wrinkle and firming efficacy test This test recruited 60 volunteers aged 25-55 with sensitive skin in each group. The participants were randomly divided into 6 groups and given the compositions of Comparative Examples 1-4, Examples 1 and 2, respectively. After 7 and 14 days of use, the anti-wrinkle and firming effects were tested using a cutometer device. The results are as follows: Table 2 - Results of Subjects Using the Product for 7 Days and 14 Days

[0048] In summary, the test data sufficiently demonstrates that Examples 1 and 2 have superior anti-wrinkle and firming effects compared to Comparative Examples 1-4. This indicates that polyglycosides... ® Microsphere-shaped PDRN, Polyglycan ® Mitochondrial PDRN has anti-wrinkle and firming effects, and urolithiasis A can promote polyglycoside A ® Microsphere-shaped PDRN, Polyglycan ® The mitochondrial PDRN composition provides better anti-wrinkle and firming effects, and has an anti-aging effect.

[0049] 5. Degradation capacity test of the composition for repairing mitochondrial mtDNA damage and anti-aging The compositions for repairing mitochondrial mtDNA damage and anti-aging prepared in Examples 1 and 2 were dispersed in 0.9% NaCl aqueous solution and left to stand at 32°C for 24 h to simulate the human body environment. Under laser irradiation, the Tyndall effect was used to see if the composition was degraded. If the Tyndall effect was present, it indicated that the spherical structure was still intact; if not, it indicated that the spherical structure had been degraded.

[0050] The results are as follows Figure 4 As shown, compared to Example 1, the composition with added urolithin A remained intact for 24 hours in physiological saline, reaching 88%, indicating that the urolithin A alcohol-protected group can maintain a spherical structure. This result is consistent with the previous human test results, showing that urolithin A can protect against polyoxin. ® Microsphere-shaped PDRN, Polyglycan ® The mitochondrial PDRN composition plays a synergistic role, primarily because urolithin A forms a hydrogen bond network with the microsphere carrier through its polyphenolic hydroxyl groups, enhancing the cross-linking density of the wall material. This results in the spherical structure maintaining 88% integrity in physiological saline for 24 hours, significantly improving the physiological stability of the microspheres and prolonging the sustained-release time of the active ingredients. Urolithin A activates mitophagy to clear damaged mitochondria, while PDRN repairs DNA damage and promotes cell proliferation, forming a closed loop of "clearance-repair." Urolithin A induces mitochondrial biogeneration to enhance cellular energy metabolism, while PDRN provides nucleotide raw materials to support genetic material synthesis, achieving a dual-dimensional anti-aging effect of "energy + structure." Microsphere technology simultaneously delivers the two active ingredients to the dermis, exerting a targeted synergistic effect.

[0051] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of this invention.

Claims

1. A composition for repairing mitochondrial mtDNA damage and anti-aging, characterized in that, By weight, it comprises: 0.01 to 0.5 parts acrylate / C10-30 alkanol acrylate crosspolymer, 0.05 to 10.0 parts microsphere PDRN, 0.01 to 10.0 parts mitochondrial PDRN, 0.01 to 2 parts inositol, 0.01 to 2.5 parts skin feeler, 1 to 30 parts moisturizer, and 0.001 to 0.2 parts anti-inflammatory and soothing ingredients, with the balance being water.

2. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1, characterized in that, It also includes 0.001 to 0.2 parts of urolithiasis A.

3. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1 or 2, characterized in that, The skin-feeling agent is mannitol and / or dextran.

4. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1 or 2, characterized in that, The moisturizer is any one or more of butylene glycol, glycerin, and 1,2-pentanediol.

5. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1 or 2, characterized in that, The anti-inflammatory and soothing ingredients are heparin sodium and / or asiaticoside.

6. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1 or 2, characterized in that, The microsphere-shaped PDRN is a polyglycoside. ® Microsphere-modified PDRN, wherein the mitochondrial PDRN is polyglycoside. ® Mitochondrial PDRN.

7. The composition for repairing mitochondrial mtDNA damage and anti-aging according to claim 1 or 2, characterized in that, By weight, it includes: 0.01~0.5 parts of acrylate / C10-30 alkanol acrylate crosspolymer, and 0.05~10.0 parts of polyglycoside. ® Microsphere-sized PDRN, 0.01~10.0 parts of polyglycoside ® Mitochondrial PDRN, 0.01-2 parts inositol, 0.01-2.5 parts mannitol, 0.2-7 parts butylene glycol, 0.2-7 parts glycerol, 0.6-16 parts 1,2-pentanediol and 0.001-0.2 parts sodium heparin, with the balance being water.

8. A method for preparing the composition for repairing mitochondrial mtDNA damage and anti-aging according to any one of claims 1 to 7, characterized in that, The acrylate / C10-30 alkanol acrylate crosspolymer and a portion of the moisturizer are dissolved in water at 70-85°C. Skin feeler and anti-inflammatory soothing ingredients are added and mixed well. The mixture is then cooled to 30-55°C, and microsphere-based PDRN, mitochondrial PDRN, inositol, the remaining moisturizer, and water are added and mixed well to obtain a composition for repairing mitochondrial mtDNA damage and anti-aging.

9. The use of the composition for repairing mitochondrial mtDNA damage and anti-aging according to any one of claims 1 to 7 in the preparation of anti-aging products.

10. A mitochondrial damage repair agent, characterized in that, A composition containing any one of claims 1 to 7 for repairing mitochondrial mtDNA damage and anti-aging.

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