Polypeptide-wrapped anti-wrinkle composition containing camellia sinensis extract and application thereof

Abstract

The application discloses a polypeptide-wrapped anti-wrinkle composition containing camellia sinensis leaf extract and application thereof, which comprises component A, component B and component C; the component A comprises palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, polyol A and caprylic / capric triglyceride; the component B comprises PPG-13-decyltetradecin polyether-24 and polyol B; and the component C comprises water and camellia sinensis leaf extract. The application firstly solves the solubility problem of palmitoyl peptides, and secondly solves the compatibility problem of camellia sinensis leaf extract and palmitoyl peptides by low-energy emulsification technology, so that a transparent flowing liquid is obtained, which is more stable, safer, more effective, more convenient to apply, can promote collagen production and inhibit the activity of elastase, and better achieves the effects of improving wrinkles and delaying aging.

Description

Technical Field

[0001] This invention relates to the field of skincare technology, specifically to a polypeptide-encapsulated anti-wrinkle composition containing red camellia extract and its application. Background Technology

[0002] Skin aging is a physiological or pathological phenomenon characterized by degenerative changes in skin morphology and function caused by the combined effects of multiple factors. It is generally influenced by both internal and external factors. External factors are primarily environmental, such as damage from ultraviolet radiation and blue light, which leads to the production of matrix metalloproteinases and the degradation of various skin proteins. Internal factors mainly refer to the degradation of the extracellular matrix (ECM) of skin cells with increasing age, especially the continuous loss of collagen in the dermis, resulting in wrinkles, loss of elasticity, and sagging skin. There are many types of collagen, with type I collagen being the most abundant structural protein, accounting for approximately 80-85%. Sufficient collagen can maintain plump and elastic skin.

[0003] Bioactive peptides, also known as functional peptides, are recognized as highly effective and safe active ingredients for preventing and combating skin aging, and are widely used in the beauty and skincare field, such as for anti-aging skincare. Peptides are a class of compounds with molecular structures between amino acids and proteins, formed by the dehydration condensation of two or more amino acids to form peptide bonds. Each active peptide has a unique structure and function, with a relatively clear mechanism of action and target on the skin, and significant efficacy is achieved even at concentrations in the ppm range. Currently, the peptides involved in the cosmetics industry are mainly small-molecule active peptides with anti-wrinkle, firming, and soothing effects, such as tripeptides, tetrapeptides, pentapeptides, and hexapeptides. With the continuous increase in consumer demand, peptides have great potential in the beauty and skincare field.

[0004] Palmitoyl-modified small peptides are among the most commonly available peptides on the market, typically existing as hydrochloride or acetate salts in crystalline solid form. However, these peptides present challenges in product development and application, primarily due to poor solubility. They are poorly soluble in water and oil, exhibiting some alcohol solubility, but also showing selectivity with different types of polyols. For example, palmitoyl tripeptide-5 has a solubility of 30 mg / mL in dimethyl sulfoxide and 30 mg / mL in ethanol, but both dimethyl sulfoxide and ethanol are unsuitable for conventional cosmetic development as they can easily cause skin allergies. Insufficient dissolution of palmitoyl peptides can also lead to precipitation, reducing their efficacy and affecting product stability, thus limiting their application in practical formulation development.

[0005] CN 116370331 A discloses a polypeptide anti-wrinkle composition and its application, comprising carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline, which has antioxidant, free radical scavenging, and collagen synthesis promoting effects, and can be used in various formulations of cosmetics. However, it has the following problem: this invention does not consider the solubility of different peptides, their compatibility, and the skin permeability of the peptides.

[0006] CN 117379335 A discloses a polypeptide liposome composition and its application, comprising acetyl heptapeptide-4, palmitoyl tripeptide-8, oligopeptide-1, phospholipids, PPG-13-decyltetradecyl alcohol polyether-24, and glycerol. The phospholipids are used to encapsulate the polypeptides via a high-pressure homogenization process, resulting in a product with high stability and easy transdermal absorption. However, the peptide efficacy involved in this patent primarily focuses on repair and soothing. Furthermore, the preparation process for encapsulating liposomes in this patent requires high pressure and multiple homogenization processes, which are complex, time-consuming, and energy-intensive. The resulting liposomes are emulsion-like and non-transparent, making them unsuitable for development of transparent products.

[0007] CN201910957516.8 discloses an anti-aging matrix composed of the following components in weight percentage: 1,2-pentanediol 3-5%, acetyl hexapeptide-8 10-20%, palmitoyl tripeptide-1 5-7%, palmitoyl tetrapeptide-7 5-8%, acetyl tetrapeptide-2 1-5%, palmitoyl tripeptide-5 3-6%, pepper seed extract 0.01-0.1%, butylene glycol 10-20%, propylene glycol 5-10%, inositol 1-5%, goldenrain fruit extract 0.05-0.1%, with the balance being deionized water. However, this patent only considers the anti-aging efficacy of the ingredients and the solubility of the peptides. Furthermore, the polyols used to dissolve the peptides have a high addition amount, resulting in high cost and insufficient solubility for the peptides. It also fails to consider peptide encapsulation and penetration, leading to low peptide bioavailability.

[0008] CN109846787A discloses a composition and its application in the preparation of cosmetics for repairing the physiological and / or biochemical structure of the skin. The composition provided by this invention comprises any two or more components selected from palmitoyl oligopeptide, flaxseed extract, hydrolyzed collagen, yarrow extract, fructose, and inulin. It utilizes a combination of multiple active ingredients to achieve skin repair and barrier function enhancement; however, its preparation method mainly involves conventional cosmetic preparation processes such as dissolution and mixing, without considering the encapsulation of palmitoyl peptides or the particle size of the composition. Although ethylhexylglycerin and 1,2-hexanediol are used to assist in the dissolution of palmitoyl peptides, effective encapsulation of active ingredients such as palmitoyl peptides is not performed. Unencapsulated active ingredients have molecular sizes and morphologies that are not conducive to effective skin penetration and absorption. This leads to reduced efficacy of the cosmetic; even if the formula contains highly effective repairing ingredients, if they cannot be fully absorbed and utilized by the skin, the expected results cannot be achieved. It also failed to mention the control of particle size in the composition; particle size directly affects the texture, stability, and skin absorption of the product; excessively large particle size results in a rough texture that is difficult to apply; while excessively small particle size may increase the difficulty and cost of preparation. Its mixing method relies on simple mechanical stirring, which cannot evenly disperse all active ingredients into the matrix, especially when there are interactions or differences in solubility between active ingredients. Summary of the Invention

[0009] Therefore, the purpose of this invention is to provide a peptide-encapsulated anti-wrinkle composition containing red camellia extract and its application. Firstly, the solubility problem of palmitoyl peptide is solved. Secondly, by encapsulating the palmitoyl peptide using low-energy emulsification technology, the compatibility problem between red camellia extract and palmitoyl peptide is resolved, resulting in a transparent, flowing liquid that is not only more stable and safer, but also more effective and easier to apply. Furthermore, it promotes collagen production and inhibits elastase activity, thus better achieving the effects of improving wrinkles and delaying aging.

[0010] The technical solution provided by this invention is as follows:

[0011] A polypeptide-encapsulated anti-wrinkle composition containing camellia extract, comprising component A, component B, and component C;

[0012] Component A includes palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, polyol A, and caprylic / capric triglyceride.

[0013] Component B includes PPG-13-decyltetradecyl alcohol polyether-24 and polyol B;

[0014] Component C includes water and red camellia extract.

[0015] The mass ratio of component A, component B, and component C is 1:(1-4):(1-6).

[0016] Preferably, the mass ratio of component A, component B, and component C is (13-18):(36-44):(38-50).

[0017] The polyol A comprises ethylhexylglycerol and 1,2-hexanediol. The mass ratio of ethylhexylglycerol to 1,2-hexanediol is between 1:1 and 6:1.

[0018] Preferably, the mass ratio of ethylhexylglycerol to 1,2-hexanediol is between 3:1 and 6:1.

[0019] The polyol B includes glycerol and butylene glycol. The mass ratio of glycerol to butylene glycol is between 1:1 and 7:1.

[0020] Preferably, the mass ratio of glycerol to butylene glycol is between 2.5:1 and 7:1.

[0021] As one embodiment of the present invention, the polypeptide-encapsulated anti-wrinkle composition containing red camellia extract comprises the following components by mass fraction:

[0022]

[0023] The polyol A comprises ethylhexylglycerol and 1,2-hexanediol. The mass ratio of ethylhexylglycerol to 1,2-hexanediol is between 1:1 and 6:1.

[0024] Preferably, the mass ratio of ethylhexylglycerol to 1,2-hexanediol is between 3:1 and 6:1.

[0025] The polyol B includes glycerol and butylene glycol. The mass ratio of glycerol to butylene glycol is between 1:1 and 7:1.

[0026] Preferably, the mass ratio of glycerol to butylene glycol is between 2.5:1 and 5:1.

[0027] As one embodiment of the present invention, the polypeptide-encapsulated anti-wrinkle composition containing red camellia extract comprises the following components by mass fraction:

[0028]

[0029] Preferably, the peptide-encapsulated anti-wrinkle composition containing camellia extract comprises the following components in the indicated mass fractions:

[0030]

[0031] The present invention also provides a method for preparing the peptide-encapsulated anti-wrinkle composition containing camellia extract as described above, comprising the following steps:

[0032] S1. Mix palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol and 1,2-hexanediol until homogeneous, add caprylic / capric triglyceride and mix further until homogeneous to obtain mixture a;

[0033] S2. Mix PPG-13-decyltetradecyl alcohol polyether-24, glycerol, and butanediol to form mixture b.

[0034] S3. Stir and mix water and red camellia extract evenly to form mixture c;

[0035] S4. Under stirring, add mixture a to mixture b and stir evenly, then add mixture c and stir (10-30 min) to obtain the peptide-encapsulated anti-wrinkle composition containing red camellia extract.

[0036] In step S4, the heating temperature is 50-65℃.

[0037] The time for adding mixture a to mixture b is 10-15 minutes.

[0038] The preparation method of red camellia extract refers to the article "Study on the anti-aging effects of red camellia flower and leaf extract in cosmetics" published in "Chinese Wild Plant Resources" and the article "Study on the efficacy of camellia flower extract" published in "Science and Technology Square".

[0039] The key features of this invention are:

[0040] The two palmitoyl-modified peptides are palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7, both of which belong to the signaling peptide class. Palmitoyl tripeptide-5, also known as a collagen peptide, promotes the production of various collagen proteins in the extracellular matrix by stimulating TGF-β (transforming growth factor), enhancing cell activity, maintaining skin elasticity, reducing wrinkles, and making the skin look younger. Palmitoyl tetrapeptide-7 is a fragment of immunoglobulin G that can reduce the secretion of IL-6 by keratinocytes after UVB exposure, thereby inhibiting the inflammatory process caused by extracellular matrix degradation, stimulating the synthesis of elastin, fibronectin, glycosaminoglycans, and type I, III, and IV collagen, improving skin texture and combating skin aging.

[0041] Red camellia extract is not only rich in flavonoid polyphenols that can scavenge free radicals and have excellent antioxidant effects, but it also contains a variety of small molecule peptides that can provide nutrition to the skin and resist skin aging.

[0042] Combining small molecule peptides with different mechanisms of action with naturally derived active ingredients like red camellia extract can create a synergistic effect, better meeting consumer needs and achieving superior anti-wrinkle efficacy. However, the varying solubility of the components and the complex composition of red camellia extract lead to poor stability and reduced efficacy in directly mixed compositions. Therefore, this invention utilizes low-energy emulsification technology to encapsulate palmitoyl peptides, protecting them from environmental damage while enhancing their penetration and release into the skin. The encapsulated palmitoyl peptides are then rationally proportioned with natural active ingredients such as red camellia extract to ensure synergistic effects and maximize the overall efficacy of the composition.

[0043] Compared with the prior art, the specific innovations of this invention are reflected in the following aspects:

[0044] 1. This invention significantly improves the solubility of palmitoyl-modified small molecule peptides (such as palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7) by carefully selecting and rationally combining different solvents, so that a high content of small molecule peptides can be added to the composition while maintaining excellent stability and avoiding precipitation.

[0045] 2. Furthermore, by employing low-energy emulsification technology, not only was the compatibility issue of directly mixing red camellia extract and palmitoyl small molecule peptides resolved, but a nano-sized composition was also successfully obtained. This innovation significantly improves the skin permeability of small molecule active peptides, resulting in higher bioavailability. The encapsulation process of this invention is green, environmentally friendly, low-energy, and simple to operate, making it highly suitable for large-scale production and achieving the dual goals of high efficiency and environmental protection. In addition, the composition is in aqueous solution form and does not contain preservatives, but through the clever combination of polyols, a good preservative effect is also achieved.

[0046] 3. After rigorous testing and verification, the peptide-encapsulated anti-wrinkle composition of this invention exhibits excellent stability. The content of encapsulated active peptides remains stable, the particle size is stable and narrow, and it has successfully passed the preservative challenge test. More importantly, it can effectively promote collagen synthesis and inhibit the activity of matrix metalloproteinases, thereby further enhancing its anti-wrinkle efficacy. Therefore, this highly effective ingredient can be widely used in various cosmetic types such as liquids, serums, creams, and lotions, satisfying consumers' eternal pursuit of anti-aging and maintaining a youthful appearance.

[0047] 4. This invention ingeniously combines small molecule peptide anti-aging ingredients with the rich skin-active ingredients in the plant red camellia, and forms a more stable and effective polypeptide anti-wrinkle composition through low-temperature emulsification technology, successfully overcoming the difficulties in formula application.

[0048] 5. Encapsulated active ingredients can more easily penetrate the skin barrier and reach deeper layers to exert their effects. The surface of the encapsulation has a lipid structure similar to that of the skin, allowing for better integration and penetration. Simultaneously, encapsulation technology can control the release rate of active ingredients, achieving a continuous and stable output of efficacy. Encapsulation technology allows for precise control of the particle size of active ingredients, thereby optimizing the product's texture, stability, and skin absorption. Attached Figure Description

[0049] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0050] Figure 1 This is a sample image of Example 1;

[0051] Figure 2 This is a particle size distribution diagram at room temperature for Example 1;

[0052] Figure 3 The particle size distribution of the peptide anti-wrinkle composition in Example 1 is shown in the diagram (under different stability testing conditions). Detailed Implementation

[0053] The present invention will be described in detail below with reference to embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0054] Preparation Example 1

[0055] For the preparation process of red camellia extract, please refer to the article "Study on the anti-aging effects of red camellia flower and leaf extract in cosmetics" published in "Chinese Wild Plant Resources" and the article "Study on the efficacy of camellia flower extract" published in "Science and Technology Square".

[0056] Examples 1-6

[0057] Examples 1-6 provide a polypeptide anti-wrinkle composition and its preparation method.

[0058] 1. Specific information on components and their contents is shown in Table 1.

[0059] Table 1 Units (mass percentage: %)

[0060]

[0061] The preparation methods of Examples 1-3 include the following steps:

[0062] 1. Mix palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol and 1,2-hexanediol until the powder is completely dissolved, then add caprylic / capric triglyceride and mix evenly to form mixture a;

[0063] 2. Mix PPG-13-decyltetradecyl alcohol polyether-24, glycerol, and butanediol until homogeneous, and this mixture is called mixture b.

[0064] 3. Mix water and red camellia extract thoroughly to form mixture c;

[0065] 4. In a 65℃ water bath, while maintaining stirring, add mixture a to mixture b at a uniform speed over 15 minutes, stir until homogeneous, and then slowly add mixture c in the same manner, stirring for 10 minutes.

[0066] 5. Stir the above mixture and cool it to room temperature, then filter to obtain a homogeneous and transparent composition.

[0067] In step 1: Palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol, and 1,2-hexanediol are mixed and stirred until the powder is completely dissolved. Ethylhexylglycerol and 1,2-hexanediol act as solubilizers, helping the peptides to dissolve more fully in the system.

[0068] In step 4, during the mixing process: mixture a is slowly added to mixture b, followed by mixture c, and then stirred. Both temperature and the slow addition method affect the encapsulation effect. A suitable temperature helps the components maintain good fluidity, allowing for more uniform mixing. In particular, the surfactant PPG-13-decyltetradecyl alcohol polyether-24 will gel if the temperature is too low, but will reduce the activity content of the active peptides if the temperature is too high. Slow addition avoids localized high concentrations or rapid reactions that could damage the encapsulation structure or create unevenness. In mixture b, PPG-13-decyltetradecyl alcohol polyether-24 acts as the main surfactant, while glycerol and butanediol act as co-surfactants, helping to reduce the resistance of the emulsifier during self-assembly at the oil-water interface. The interaction between mixture b and the components in mixture a further refines and stabilizes the encapsulation structure, forming an oil-in-alcohol mixture. The slow addition of mixture c then forms an oil-in-water mixture, reinforcing or optimizing the already preliminarily formed encapsulated active peptide structure, making it more tightly and stably encapsulated, thus achieving a stable and narrow particle size range.

[0069] The preparation methods of Examples 4-6 include the following steps:

[0070] 1. Mix palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol and 1,2-hexanediol until the powder is completely dissolved, then add caprylic / capric triglyceride and mix evenly to form mixture a;

[0071] 2. Mix PPG-13-decyltetradecyl alcohol polyether-24, glycerol, and butanediol until homogeneous, and this mixture is called mixture b.

[0072] 3. Mix water and red camellia extract thoroughly to form mixture c;

[0073] 4. In a 55℃ water bath, while maintaining stirring, slowly add mixture a to mixture b at a uniform speed over 10 minutes, stirring until homogeneous. Then slowly add mixture c and stir for 30 minutes.

[0074] 5. Stir the above mixture and cool it to room temperature, then filter to obtain a homogeneous and transparent composition.

[0075] Based on the composition, dosage, and preparation process described above, the compositions obtained in Examples 1-6 are all transparent flowing liquids, and the measured particle sizes are all in the range of 30-200 nm.

[0076] Comparative Example 1

[0077] The preparation process of this comparative example is the same as that of Example 1. The only difference is that red camellia extract is not added to the components. The resulting composition is a transparent flowing liquid with a particle size (D50) of 40 nm. However, the anti-wrinkle effect of this composition is not as good as that of Example 1.

[0078] Comparative Example 2

[0079] The preparation process of this comparative example is the same as that of Example 1, except that the ratio of component A: component B: component C is 3:0.5:2. Using this ratio, a transparent and uniform composition cannot be prepared, which does not meet the application requirements.

[0080] Comparative Example 3

[0081] Publication patent 202211695584.X relates to a microemulsion and its preparation method, comprising an emulsifier, a polyol, an oil phase, and water. The emulsifier is PPG-13-decyltetradecyl alcohol polyether-24; the polyol is glycerol and / or diglycerol, which may also be loaded with active ingredients. Using the preparation method related to oil-soluble active ingredients in this patent, experiments revealed that neither caprylic / capric triglyceride nor camellia seed oil could dissolve palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7, nor could a transparent and homogeneous composition be obtained.

[0082] Comparative Example 4

[0083] The preparation process of this comparative example is the same as that of Example 1. The only difference is that butanediol is not added to the components. The resulting composition is a transparent flowing liquid with a particle size (D50) of 60 nm. However, obvious stratification occurred during the stability test at high temperatures of 40°C and 48°C, and the stability test was not passed.

[0084] Comparative Example 5

[0085] The preparation process of this comparative example is the same as that of Example 1. The only difference is that the mass ratio of glycerol to butylene glycol is 1:2, and the specific addition amounts are 7% and 14%, respectively. The resulting composition is a semi-transparent, slightly whitish, bluish liquid with a particle size (D50) of 250.28 nm. During the stability test, it showed fogging at low temperatures of 3°C and -18°C, and oil precipitation. The stability test was not passed.

[0086] Comparative Example 6

[0087] The preparation process of this comparative example is the same as that of Example 1. The only difference is that the ratio of PPG-13-decyltetradecyl alcohol polyether-24 to caprylic / capric triglyceride is 1:2, and the addition amounts are 6% and 12%, respectively, to obtain a semi-permeable emulsion composition with a particle size (D50) greater than 1000 nm.

[0088] Comparative Example 7

[0089] The preparation process of this comparative example is the same as that of Example 1. The only difference is that ethylhexylglycerin is not added to the components. The resulting composition is a nearly transparent flowing liquid with a particle size (D50) of 50 nm. However, after being placed at room temperature for 1 week, solid particles precipitated out. After testing, it was confirmed that the precipitates were palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7. The reason for this was that the palmitoyl peptides were not fully dissolved.

[0090] Comparative Example 8

[0091] The preparation process of this comparative example is the same as that of Example 1. The only difference is that ethylhexylglycerol is replaced with tocopherol acetate in the same proportion. In step 1, a transparent and uniform mixture a cannot be obtained. There is white powder in the semi-transparent liquid, which was identified as palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7 by analysis, indicating that tocopherol acetate cannot dissolve palmitoyl peptides well.

[0092] Comparative Example 9

[0093] The preparation process of this comparative example is the same as that of Example 1. The only difference is that ethylhexylglycerol is replaced with 1,2-pentanediol in the same proportion. The resulting composition is a semi-transparent bluish flowing liquid. After being left at room temperature for 1 week, solid particles precipitated at the bottom. After testing, the precipitates were confirmed to be palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7. The reason for this was that 1,2-pentanediol does not have good solubility for palmitoyl peptides.

[0094] Comparative Example 10

[0095] The preparation process of this comparative example is the same as that of Example 1. The only difference is that ethylhexylglycerol is replaced with diisopropyl adipate in the same proportion. In step 1, a transparent and uniform mixture a cannot be obtained. There are white flocculent substances in the semi-permeable liquid. After analysis, they were identified as palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7, indicating that diisopropyl adipate cannot dissolve palmitoyl peptides well.

[0096] Comparative Example 11

[0097] The composition and proportions of this comparative example are the same as those of Example 1, the only difference being the preparation method of the composition.

[0098] The specific preparation method for this comparative example is as follows:

[0099] At 65°C, water, glycerol, butylene glycol, camellia extract, palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerin, 1,2-hexanediol, acetyl hexapeptide-8, PPG-13-decyltetradecyl alcohol polyether-24, and caprylic / capric triglyceride were added sequentially under stirring. The mixture was stirred until homogeneous, but a uniform composition could not be obtained.

[0100] Comparative Example 12

[0101] The composition and proportions of this comparative example are the same as those of Example 1, the only difference being the preparation method of the composition.

[0102] In step 4 of the preparation process, mixture b and mixture c are added to mixture a simultaneously and stirred until they are homogeneous, finally obtaining a semi-transparent bluish composition. The composition has a wide particle size range and poor uniformity.

[0103] Comparative Example 13

[0104] The composition and proportions of this comparative example are the same as those of Example 1, the only difference being in step 1.

[0105] Palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol, 1,2-hexanediol, and caprylic / capric triglyceride are simultaneously mixed and dispersed as mixture a. Using this method, a transparent and homogeneous mixture a cannot be obtained, which ultimately affects the transparency of the composition and causes the palmitoyl peptide to not dissolve sufficiently.

[0106] Performance Test Example 1: Stability Study of the Peptide Anti-wrinkle Composition

[0107] The compositions prepared in Examples 1-6 were subjected to a 2-month stability study, and the results are shown in Table 2.

[0108] Table 2. Results of stability study of peptide anti-wrinkle compositions

[0109]

[0110]

[0111] As can be seen from the results in Table 2, the compositions under different conditions all maintained a uniform and transparent liquid state, and no precipitation, fogging, or stratification occurred, indicating that the stability test was passed.

[0112] Performance Test 2: Particle Size Test of the Peptide Anti-Wrinkle Composition

[0113] The particle size of the compositions was tested using a Bettersize 3000 particle size analyzer. The particle size test results for Examples 1-6 are shown in Table 3. Taking Example 1 as a representative example, the initial particle size test results and the particle size test results after a 2-month stability study are shown in the figure. Figure 2 and Figure 3 As shown.

[0114] Table 3. Particle size test results of Examples 1-6

[0115]

[0116] The results show that the particle size of the peptide-encapsulated anti-wrinkle composition containing camellia extract obtained in this invention is within 200 nm, with most particles within 100 nm. After a two-month stability study, the particle size of the peptide anti-wrinkle composition remained within 200 nm, with most particles within 100 nm, and the particle size variation under different conditions was small, indicating that the composition is very stable.

[0117] Performance Test 3: Determination of Peptide Content in the Peptide Anti-wrinkle Composition

[0118] Taking Example 1 as a representative example, the content of some small molecule peptides in Example 1 was determined after the stability study was completed.

[0119] The specific steps are as follows: High-performance liquid chromatography (HPLC) with a UV detector was used. Reagents included acetonitrile (chromatographic grade), phosphoric acid (analytical grade), triethylamine (analytical grade), potassium dihydrogen phosphate, palmitoyl tripeptide-5, and palmitoyl tetrapeptide-7. Blank solution, standard solution, and test solution were precisely injected into the HPLC system, and chromatograms were recorded. The contents of palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7 were calculated using the external standard method, and the purity of the test sample was calculated using the area normalization method. The results are shown in Table 4. The results show that after stability testing, the contents of palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7 encapsulated in the peptide anti-wrinkle composition remained stable under all conditions.

[0120] Table 4. Determination of the content of palmitoyl tripeptide-5 and palmitoyl tetrapeptide-7 in the peptide anti-wrinkle composition.

[0121] room temperature 200.20 200.89 40℃ 198.74 198.16 3℃ 197.69 200.77 -10℃ 199.51 198.79

[0122] Performance Test 4: Transdermal Absorption Test of the Peptide Anti-wrinkle Composition

[0123] Transdermal absorption in cosmetics refers to the process by which functional ingredients in cosmetics act on the skin surface or penetrate the epidermis or dermis according to the product's effectiveness, accumulating and exerting their effects at that site. Currently, the most commonly used in vitro method for detecting transdermal absorption of cosmetic ingredients is the diffusion cell method. Human or animal skin is attached to a diffusion cell, and the receiving solution is collected periodically. The content of the active ingredient is determined using appropriate analytical methods, and the transdermal rate is calculated to evaluate the penetration characteristics of the chemical substance.

[0124] This performance test used a transdermal absorption diffusion cell, selecting abdominal skin of Bama miniature pigs as the ex vivo skin. HPLC was used to detect the target active ingredient, ultimately obtaining the release and cumulative amount of the active ingredient at different time points. Taking Example 1 and Comparative Example 9 as representative examples, the sample volume was 0.5 ml of stock solution, with sampling points at 8 hours. The transdermal absorption results are shown in Table 5.

[0125] Table 5. Transdermal absorption test results of palmitoyl tripeptide-5 in the peptide anti-wrinkle composition.

[0126]

[0127] The results show that when the sample was taken for testing at 8 hours, the release amount of palmitoyl tripeptide-5 in Example 1 exceeded that of Comparative Example 9, and the amount retained in the skin was much greater than that of Comparative Example 9. This indicates that the polypeptide anti-wrinkle composition prepared by the present invention can help the active ingredients be released slowly and the transdermal absorption capacity is greatly improved.

[0128] Performance Test 5

[0129] Preservative challenge test of peptide anti-wrinkle composition:

[0130] The samples from Example 1 were tested according to the General Principles of Microbial Testing Methods, Total Colony Count, and Test Methods for Molds and Yeasts in Chapter 5 of the "Cosmetic Safety Technical Specifications" (2015 Edition). The test results should meet the limit requirements in the "Cosmetic Safety Technical Specifications" (2015 Edition). The preservative challenge test adopted a single-strain contamination method, that is, the test sample was artificially contaminated with a bacterial suspension of each test strain, and the number of surviving bacteria was measured at regular intervals. The challenge of the cosmetic preservative system was judged based on the changes in the number of surviving bacteria. The test methods and judgment criteria for this preservative challenge experiment were formulated with reference to the United States Pharmacopeia (USP51), the European Pharmacopoeia (EP7.0), and the CTFA method. A single inoculation method was adopted, with inoculation on day 0, and results were tested at 7, 14, 21, and 28 days after inoculation. The experimental results confirmed that the samples passed the preservative challenge and had good preservative efficacy. The test results are shown in Tables 6 and 7.

[0131] Table 6. Results of sterility testing

[0132]

[0133] Table 7 Corrosion Challenge Test Results

[0134] bacteria <10 <10 <10 <10 fungi 330 60 <10 <10 .

[0135] Performance Test 6

[0136] Combining palmitoyl peptide and red camellia extract in a proper way can better promote collagen synthesis, thereby achieving better anti-wrinkle effects.

[0137] The specific test results are shown in Table 8.

[0138] In vitro efficacy testing of peptide anti-wrinkle compositions: testing for promoting type I collagen production.

[0139] The peptide-encapsulated anti-wrinkle compositions prepared in Examples 1-6 and Comparative Example 1 were tested for their ability to promote type I collagen. The testing method followed the group standard T / SHRH 031—2020 "Testing of Firming and Anti-wrinkle Efficacy of Cosmetics - Determination of Type I Collagen Content in In Vitro Fibroblasts" published by the Shanghai Daily Cosmetics Industry Association. HSF cells in the logarithmic growth phase were seeded in 3cm diameter cell culture dishes or 6-well cell culture plates. After the cells reached 80% confluence, different concentrations of the sample were added to the sample groups for culture. After replacing the culture medium with fresh medium, the cells were cultured for another 24 hours. The cell supernatant was collected, and the type I collagen content was detected using an ELISA kit. Higher content indicates stronger anti-wrinkle ability.

[0140] Relative content of type I collagen = T / C × 100%

[0141] In the formula:

[0142] T—The average content of type I collagen in the test substance;

[0143] C—Number of type I collagen contents (blank / solvent control)

[0144] Table 8 Results of determination of relative content of type I collagen

[0145] Normal control / 100.00％ Positive control 50 ng / mL 126.33％ Example 1 0.01％ 119.35％ Example 2 0.01％ 117.28％ Example 3 0.01％ 116.97％ Example 4 0.01％ 124.59％ Example 5 0.01％ 130.86％ Example 6 0.01％ 123.22％ Comparative Example 1 0.01％ 110.77％

[0146] The specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A polypeptide-encapsulated anti-wrinkle composition containing camellia extract, characterized in that, Includes component A, component B, and component C; Component A includes palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, polyol A, and caprylic / capric triglyceride; wherein polyol A includes ethylhexylglycerol and 1,2-hexanediol. Component B includes PPG-13-decyltetradecyl alcohol polyether-24 and polyol B; said polyol B includes glycerol and butanediol. Component C includes water and red camellia extract; The peptide-encapsulated anti-wrinkle composition containing red camellia extract, by mass fraction, comprises: Palmitoyl tripeptide - 50.001 - 0.1% Palmitoyl tetrapeptide - 70.001 - 0.1% Polyol A5-15% Caprylic / Capric Triglycerides 5-10% PPG-13-Decyltetradecyl alcohol polyether-245-20% Polyol B15-35% Water 20-50% Red camellia extract 0.1-10%; The aforementioned peptide-encapsulated anti-wrinkle composition containing camellia extract is prepared by a method comprising the following steps: S1. Mix palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, ethylhexylglycerol and 1,2-hexanediol until homogeneous, add caprylic / capric triglyceride and mix further until homogeneous to obtain mixture a; S2. Mix PPG-13-decyltetradecyl alcohol polyether-24, glycerol, and butanediol to form mixture b. S3. Stir and mix water and red camellia extract until homogeneous to form mixture c; S4. Under heating and stirring, add mixture a to mixture b and stir evenly, then add mixture c and continue stirring to obtain the peptide-encapsulated anti-wrinkle composition containing red camellia extract; In step S4, the heating temperature is 50-65℃.

2. The polypeptide-encapsulated anti-wrinkle composition containing camellia extract according to claim 1, characterized in that, The mass ratio of ethylhexylglycerol to 1,2-hexanediol is between 1:1 and 6:

1.

3. The polypeptide-encapsulated anti-wrinkle composition containing camellia extract according to claim 1, characterized in that, The mass ratio of glycerol to butylene glycol is between 1:1 and 7:

1.

4. The polypeptide-encapsulated anti-wrinkle composition containing camellia extract according to claim 1, characterized in that, The mass ratio of component A, component B, and component C is 1:1-4:1-6.

5. The use of a polypeptide-encapsulated anti-wrinkle composition containing camellia extract as described in any one of claims 1-4 in the preparation of cosmetics.

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