Anti-aging composition

Abstract

The present invention discloses an anti-aging composition, which is composed of the following components by weight: 0.5 to 3 parts of Meconopsis pilosula extract; 0.0005 to 0.0025 parts of nonapeptide-1 or acetyl hexapeptide; 0.0001 to 0.0005 parts of palmitoyl hexapeptide-12; 0.005 to 0.01 parts of mulberry root extract; 0.5 to 3 parts of supramolecular schizonepeta tenuifolia extract; 0.15 to 0.5 parts of peony root bark extract; and 0.5 to 3 parts of arginine / lysine polypeptide. The novel anti-aging composition of the present invention is based on the comprehensive anti-aging solution that takes into account the epidermis, dermis, and muscle layers proposed in the early stage. It is further designed from the perspectives of epidermal barrier repair, basement membrane repair, dermal repair, and mitochondrial ROS clearance, and does not use strong active ingredients that are irritating to the skin, ensuring that the product itself will not damage the skin barrier and cause skin inflammation.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics and relates to an anti-aging composition. Background Art

[0002] As the aging population intensifies, skin aging has become a common concern in the health and cosmetics industries. Due to natural aging and external stimuli such as ultraviolet rays, pollution and other environmental issues, the demand for anti-aging products is constantly increasing. Continuously improving the efficacy of anti-aging products is of great significance.

[0003] The skin is the largest organ in the human body, and its most important function is the barrier function, which prevents external stimuli from damaging the skin and prevents the loss of water from the skin. As described in the patent application number CN202111658056.2, with natural aging, the epidermis first shows barrier damage, resulting in a decrease in skin water content, weakened proliferation of keratinocytes, thinning of the epidermis, and the appearance of fine lines on the skin. The damaged barrier leads to a decrease in the skin's protective ability, making the skin more susceptible to external damage, leading to the activation of MMPs, which degrades collagen and elastin in the dermis. At the same time, the fibroblasts in the dermis show signs of aging. Due to reasons such as mitochondrial oxygen supply and shortened telomeres of genes in the cell nucleus, the proliferation ability of fibroblasts is weakened, and the ability to secrete collagen and elastin is reduced, further exacerbating the reduction of dermal extracellular matrix substances. The dermis's support for the skin weakens with age, and the skin gradually becomes loose and sagging. In the problem of aging, due to the continuous contraction of muscles, expression lines accumulate over time, making the skin look old. Expression lines are an important factor in the skin's aging state. The realization of skin functions depends on the energy generated by respiration. Mitochondria produce a large amount of ROS while generating energy. Cell aging leads to a weakening of the ability to eliminate free radicals. Excessive ROS can cause a series of skin problems.

[0004] The skin barrier is very important to skin health, but the existence of the skin barrier will also affect the effectiveness of topical preparations and the penetration and absorption of anti-aging active substances. Therefore, in order to further improve the efficacy of anti-aging active substances, consideration of the transdermal penetration of raw materials is very critical.

[0005] In patent application number CN202111658056.2, the applicant proposed an anti-aging composition that takes into account the epidermis, dermis, and muscle and follows the anti-aging mechanism, as well as a cosmetic containing the composition. However, there are three aspects that need further optimization:

[0006] (1) The basement membrane of the skin is responsible for the riveting effect of the keratinocytes in the basal layer of the epidermis and plays a very important role in the communication of information and nutrition between the epidermis and the dermis. However, this is not mentioned in the patent application number CN202111658056.2;

[0007] (2) Beauty peptides generally refer to peptides containing no more than 10 amino acids, which are widely used in cosmetics, especially anti-aging products. The patent application number CN202111658056.2 also adds palmitoyl hexapeptide-12, which has a significant effect on promoting collagen production. However, the stability of the peptide raw material will be affected by the pH value of the formula, other active ingredients in the formula, the storage temperature of the product, etc. How to improve the stability of the peptide raw material and maximize its efficacy is not considered in the patent application number CN202111658056.2;

[0008] (3) The transdermal ability of active ingredients is an important factor in the effectiveness of cosmetics, especially anti-aging cosmetics, but it is not mentioned in the patent application number CN202111658056.2.

[0009] Therefore, in order to further improve the efficacy and ensure the anti-aging effect of the product, continuously improving the anti-aging dimension of the skin and optimizing the product stability and transdermal ability are key points in product design. Summary of the Invention

[0010] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a skin anti-aging composition, particularly for increasing the secretion of basement membrane DEJ-related proteins and collagen and mucopolysaccharide in the extracellular matrix. The present invention provides a skin anti-aging composition that forms liposomes by combining the core collagen secretion-promoting peptide raw materials, palmitoyl hexapeptide-12 and nonapeptide-1 / acetyl hexapeptide, to promote the penetration of the raw materials and enhance the effect of promoting collagen secretion by fibroblasts. To enhance the effect of the efficacy of the ingredients at the muscle level, schizonepeta is combined with propylene glycol and betaine ligands to form a supramolecular structure to promote penetration and absorption, thereby enhancing its effect in improving expression lines. Furthermore, the composition is designed from the perspectives of epidermal barrier repair, DEJ repair, dermal repair, and mitochondrial ROS clearance, still taking into account the epidermis, dermis, and muscle layers from a dermatological perspective to achieve multi-dimensional skin anti-aging.

[0011] The technical solution for achieving the above object is: an anti-aging composition, which is composed of the following components in parts by weight:

[0012]

[0013]

[0014] In the above-mentioned anti-aging composition, the Meconopsis pilosula extract has the effect of improving cell vitality.

[0015] In the above-mentioned anti-aging composition, the Meconopsis extract is obtained by extracting Meconopsis plants, and the Meconopsis plants are one or more of Meconopsis oleifera, Meconopsis yunnanensis, Meconopsis racemosa and Meconopsis entireta.

[0016] In the above-mentioned anti-aging composition, the extraction process of the Meconopsis plant is one or more of water extraction, alcohol extraction, NADES extraction technology and subcritical extraction technology.

[0017] The above-mentioned anti-aging composition, wherein the nonapeptide-1 / acetyl hexapeptide has the effect of promoting the secretion of basement membrane-associated proteins and mucopolysaccharides, and the basement membrane-associated proteins include collagen IV, collagen VII, collagen XVII and laminin.

[0018] In the above-mentioned anti-aging composition, the palmitoyl hexapeptide-12 and nonapeptide-1 are encapsulated in liposomes and used together to promote dermal fibroblasts to secrete type I and type III collagen.

[0019] In the above-mentioned anti-aging composition, the encapsulating material of the liposome encapsulating palmitoyl hexapeptide-12 and nonapeptide-1 is one or more of ginsenoside, lecithin and hydrogenated lecithin.

[0020] The above-mentioned anti-aging composition, wherein the supramolecular spilionella extract forms a supramolecular structure with propylene glycol and betaine, and the supramolecular spilionella extract is used to block the Na+ ion influx channel, reduce the transmission of nerve impulse signals, and relax muscles.

[0021] In the above-mentioned anti-aging composition, the peony root bark extract and the arginine / lysine polypeptide act together to reduce the production of ROS in mitochondria.

[0022] The anti-aging composition of the present invention is designed from the perspectives of epidermal barrier repair, basement membrane DEJ repair, dermal repair, and mitochondrial ROS clearance. Based on the patent application number CN202111658056.2, it is more comprehensive and stronger in terms of dermatological anti-aging, taking into account the epidermis, dermis, and muscle levels, while also taking into account safety. It does not use strong active ingredients that are irritating to the skin, ensuring that the product itself will not damage the skin barrier and cause skin inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the liposome particle size distribution diagram of palmitoyl hexapeptide-12 and nonapeptide-1;

[0024] Figure 2 This is the result of the stability study of nonapeptide-1 in liposomes;

[0025] Figure 3 This is the result of the stability study of palmitoyl hexapeptide-12 in liposomes;

[0026] Figure 4This is a diagram showing the results of a zebrafish experiment on the neuroinhibition effect of common spinach extract;

[0027] Figure 5 This is a diagram showing the results of a zebrafish experiment on the neuroinhibition effect of supramolecular Spinach extract;

[0028] Figure 6 This is the collagen I test result chart;

[0029] Figure 7 This is the collagen III test result chart;

[0030] Figure 8 This is the collagen IV test result chart. DETAILED DESCRIPTION

[0031] In order to enable those skilled in the art to better understand the technical solution of the present invention, the specific implementation methods thereof are described in detail below with reference to the accompanying drawings:

[0032] An anti-aging composition, comprising the following components in parts by weight:

[0033]

[0034] The anti-aging composition of the present invention comprises the Meconopsis thunbergii extract which can increase the expression of HES1 and thus improve cell activity.

[0035] Experimental methods:

[0036] 1) Inoculation: 2×10 5 The cells were seeded into 6-well plates at a seeding density of 100 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ).

[0037] 2) Solution preparation: Prepare the working solution of the test substance according to a certain concentration.

[0038] 3) Drug administration: When the cell plating rate in the 6-well plate reaches 40% to 60%, the cells are divided into groups for drug administration. The dosage per well is 2 mL. Each group has 3 replicate wells and is incubated in an incubator (37° C., 5% CO 2 ) for 24 h.

[0039] 4) UVA irradiation: UVA irradiation was performed on the groups that needed irradiation according to the test group, with an irradiation dose of 30 J / cm2. After irradiation, the cells were placed in an incubator (37°C, 5% CO2) and cultured for 24 hours.

[0040] 5) Gene Expression Assay: After 24 hours of culture, cell supernatants were collected and washed twice with 1 mL / well PBS. 1 mL of RNAiso Plus was added to each well, and cells were lysed by pipetting. Samples were then collected. RNA was extracted, reverse-transcribed to cDNA, and analyzed by quantitative PCR. Results were calculated using the 2-ΔΔCT method.

[0041] Table 1, results of Meconopsis extract increasing the expression of HES1:

[0042] Sample name Average relative expression level SD P-value Increase rate BC 1.00 0.12 / / NC 0.75 0.03 0.027# / PC 2.14 0.21 0.000** 185.33％ Meconopsis extract 1.18 0.06 0.000** 57.33％

[0043] See also Figure 1 、 Figure 2 and Figure 3 The liposome particle size of palmitoyl hexapeptide-12 and nonapeptide-1 was analyzed, and the stability of palmitoyl hexapeptide-12 and nonapeptide-1 in liposomes was investigated. The specific stability method is as follows:

[0044] 1. Photothermal stability test

[0045] The liposome samples and standard solutions were placed at -15℃, 4℃, 25℃, 40℃ light and 49℃ conditions for 7 days, and the contents of nonapeptide-1 and palmitoyl hexapeptide-12 were determined to investigate the photothermal stability of the liposomes.

[0046] 2. Freeze-thaw test

[0047] The liposome samples and standard solutions were separately packed into small test tubes, frozen in the refrigerator for 24 hours, thawed in cold water, and then continued to be frozen in the refrigerator. After the samples and standards underwent four freeze-thaw cycles, the contents of encapsulated and unencapsulated nonapeptide-1 and palmitoyl hexapeptide-12 were measured to investigate the freezing stability of the liposomes.

[0048] The contents of nonapeptide-1 and palmitoyl hexapeptide-12 in the liposomes were tested by HPLC. After the liposomes were dissolved in ethanol, the samples were analyzed after sonication and centrifugation. A ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm) was used for separation. The mobile phase was 0.1% trifluoroacetic acid aqueous solution-acetonitrile as the gradient elution, and the detection was carried out at a wavelength of 215 nm using a UV detector.

[0049] from Figure 1 、 Figure 2 and Figure 3 The results show that palmitoyl hexapeptide-12 and nonapeptide-1 can promote the secretion of type I and type III collagen by dermal fibroblasts, and have the effect of repairing the dermal extracellular matrix. Using one or more of ginsenosides, lecithin, and hydrogenated lecithin as encapsulating materials to form liposomes improves the stability of palmitoyl hexapeptide-12 and nonapeptide-1.

[0050] Because peony root bark extract contains paeonol, it has the function of scavenging free radicals. In the patent application number CN202111658056.2, peony root bark extract is combined with ergothioneine, which can play the role of both superoxide dismutase and catalase in the antioxidant chain. However, due to the stability of ergothioneine in the formula and its unpleasant odor, it is difficult to use it in products without adding flavors. Therefore, considering the importance of the pleasant smell of anti-aging products, arginine / lysine peptide is used instead of ergothioneine and combined with peony root bark extract to jointly play the role of reducing ROS in mitochondria.

[0051] See also Figure 4 and Figure 5 , schisandra extract has the effect of blocking the influx of Na+ ions, reducing the transmission of nerve impulse signals and relaxing muscles. In order to further improve the penetration of schisandra extract and exert its efficacy, schisandra extract is combined with propylene glycol and betaine to form a supramolecular structure to improve its efficacy. Figure 5 and Figure 6 showed that, by comparison, the neuroinhibitory effect of supramolecular schizonepeta extract was enhanced by 74%.

[0052] Experimental Methods: A pentylenetetrazol (PTZ)-induced seizure model in zebrafish was used. Twenty-four five-day-old zebrafish embryos were exposed to a 7.5 mM PTZ solution supplemented with a spilanthus radiata extract sample in a 96-well plate. A PTZ control group was also included. After 40 minutes of exposure, the activity of each embryo was recorded over a 20-minute period using a DanioVision zebrafish behavioral recorder using EthoVision XT infrared tracking software, and statistical analysis was performed.

[0053] Table 2, formula of the anti-aging cosmetics of Examples 1 to 4:

[0054]

[0055]

[0056] Anti-aging cosmetics containing the anti-aging compositions of the present invention according to Examples 1-4 were prepared according to the anti-aging composition formulas in Table 2. The anti-aging compositions of the present invention are comprised of Meconopsis pilosa extract, nonapeptide-1 / acetyl hexapeptide, palmitoyl hexapeptide-12, mulberry root extract, supramolecular Spinach extract, Paeonia suffruticosa root bark extract, and arginine / lysine polypeptide.

[0057] See also Figure 6 、 Figure 7 and Figure 8The upgraded version of MLYAAT composition in Example 3 was tested for in vitro efficacy. The expression of collagen I, collagen III, and collagen IV on the in vitro skin model was tested. The test results of collagen I are shown in Tables 3 and Figure 6 , collagen Ⅲ test results are shown in Table 4 and Figure 7 , collagen Ⅳ test results are shown in Table 5 and Figure 8 . Figure 6 、 Figure 7 and Figure 8 BC represents the blank group, which was not stimulated or medicated; NC represents the negative group, which was stimulated but not medicated; PC (VC+VE) represents the positive group, which was given a certain concentration of vitamin C and vitamin D after the stimulation treatment as a positive control group; the upgraded version of MYAT combination is Example 3 of the present invention, also known as the upgraded MLYAAT combination.

[0058] The specific method for in vitro skin model collagen testing is as follows:

[0059] 1. Tissue Processing: Freshly obtained skin tissue was immersed in 75% alcohol for 30 seconds and then washed three times with sterile PBS buffer. After washing, the skin was cut into 24 ± 2 mm2 tissue blocks with the epidermis facing up and the dermis facing down. The blocks were placed in a culture mold. The culture mold was then transferred to a 6-well plate. 3.7 mL of culture medium was added to each well. The plates were incubated at 37°C in a 5% CO2 incubator, with the medium changed daily.

[0060] 2. Dosing: After 2 days of in vitro skin tissue culture, the tissues were treated according to the corresponding treatment conditions according to the grouping, and irradiation and dosing began. The irradiation dose was UVA (30J / cm2) and UVB (50mJ / cm2) for 4 consecutive days. Fresh culture medium was replaced after each irradiation period, and dosing was performed. The positive control (VC+VE) was administered via submerged dosing, and the test samples were administered via surface dosing. After 4 days of continuous irradiation, the ex vivo skin tissues were cultured for an additional 3 days without irradiation, during which only sample dosing was performed.

[0061] 3. Collagen fiber detection: After the administration, the skin tissue was fixed with 4% paraformaldehyde, embedded, and sliced ​​for Masson staining and Victoria blue staining. The slices were collected and photographed using a microscope. Plus image processing software for analysis.

[0062] 4. Immunohistochemistry: Perform the test according to the specific immunohistochemistry procedures.

[0063] 5. Immunofluorescence detection: Take the tissue for immunofluorescence detection and fix it with 4% paraformaldehyde. After fixation for 24 hours, take pictures with a fluorescence microscope and use Plus image processing software for analysis.

[0064] 6. Statistical Analysis: Graphs were generated using GraphPad Prism, and results are expressed as mean ± SD. Comparisons between groups were analyzed using the t-test. All statistical analyses were two-tailed. P < 0.05 was considered a significant difference, and P < 0.01 was considered a highly significant difference.

[0065] Table 3, Collagen I test results

[0066] Group Relative IOD average SD P-value Improvement rate (vs NC) BC 1.00 0.10 / / NC 0.49 0.03 0.001## / PC(VC+VE) 1.16 0.04 0.000** / MYAT upgraded version combination 1.13 0.02 0.000** 130.61％

[0067] Table 4, Collagen III test results

[0068] Group Relative IOD average SD P-value Improvement rate (vs NC) BC 1.00 0.16 / / NC 0.31 0.06 0.002## / PC(VC+VE) 0.71 0.12 0.006** / MYAT upgraded version combination 0.84 0.12 0.003** 170.97％

[0069] Table 5, Collagen IV test results

[0070]

[0071] According to Table 3-5 and Figure 6-8 The results show that Example 3, i.e., the upgraded MLYAAT combination, significantly increased the expression of collagen I and collagen III in the dermal extracellular matrix (ECM) and collagen IV in the DEJ in the in vitro skin model, and had a significant anti-wrinkle effect.

[0072] In summary, the anti-aging composition of the present invention fully considers the importance of the anti-aging dimension of the basement membrane, the stability of the peptide raw materials, and the transdermal permeability of the functional raw materials. It is designed more comprehensively and stronger from the perspectives of epidermal barrier repair, basement membrane DEJ repair, dermal repair, and mitochondrial ROS clearance. It takes into account the anti-aging of the epidermis, dermis, and muscle levels from a dermatological perspective, and takes into account safety. It does not use strong active ingredients that are irritating to the skin, ensuring that the product itself will not damage the skin barrier and cause skin inflammation.

[0073] Those skilled in the art should recognize that the above embodiments are merely intended to illustrate the present invention and are not intended to limit the present invention. As long as they are within the spirit of the present invention, any changes or modifications to the above embodiments will fall within the scope of the claims of the present invention.

Claims

1. An anti-aging composition, characterized in that By weight, it is composed of the following components: The palmitoyl hexapeptide-12 and nonapeptide-1 are encapsulated in liposomes and used together to promote the secretion of type I and type III collagen by dermal fibroblasts; In the supramolecular spilionella extract, the spilionella extract forms a supramolecular structure with propylene glycol and betaine. The supramolecular spilionella extract is used to block the Na+ ion influx channel, reduce the transmission of nerve impulse signals, and relax muscles.

2. An anti-aging composition according to claim 1, characterized in that The Meconopsis pilosula extract has the effect of improving cell vitality.

3. An anti-aging composition according to claim 1 or 2, characterized in that: The Meconopsis extract is extracted from Meconopsis plants, and the Meconopsis plants are one or more of Meconopsis oleifera, Meconopsis yunnanensis, Meconopsis racemosa and Meconopsis integrifolia.

4. An anti-aging composition according to claim 3, characterized in that The extraction process of Meconopsis plant is one or more of water extraction, alcohol extraction, NADES extraction technology and subcritical extraction technology.

5. An anti-aging composition according to claim 1, characterized in that: The nonapeptide-1 or acetyl hexapeptide has the function of promoting the secretion of basement membrane-related proteins and mucopolysaccharides. The basement membrane-related proteins include collagen IV, collagen VII, collagen XVII and laminin.

6. An anti-aging composition according to claim 1, characterized in that: The encapsulating material of the liposome encapsulating palmitoyl hexapeptide-12 and nonapeptide-1 is one or more of ginsenoside, lecithin and hydrogenated lecithin.

7. An anti-aging composition according to claim 1, characterized in that: The peony root bark extract and arginine / lysine polypeptide act together to reduce the generation of ROS by mitochondria.

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