CN116509748A - Anti-wrinkle composition and preparation method and application thereof - Google Patents

Abstract

The invention discloses an anti-wrinkle composition, a preparation method and application thereof. The anti-wrinkle composition of the present invention includes: a polypeptide that induces elastin and type I collagen synthesis with a long-term anti-wrinkle effect, an acetylcholine antagonist and a sodium ion channel inhibitor with an immediate anti-wrinkle effect, a supramolecular ionic liquid with a penetration-promoting effect, HEPES. The anti-wrinkle composition provided by the present invention starts from the different action targets of the polypeptide, combines supramolecular ionic liquid to deliver the composite polypeptide, takes into account both specimens, and achieves the anti-wrinkle effect of 1+1>2. At the same time, supramolecular ionic liquids are also components of functional active substances in cosmetics. While promoting the absorption of active substances, they can also protect the structure of active substances and play a synergistic role.

Description

A kind of anti-wrinkle composition and its preparation method and application

technical field

The invention relates to the field of cosmetics, in particular to an anti-wrinkle composition and its preparation method and application.

Background technique

The appearance of wrinkles is an important sign of skin aging. The formation of wrinkles is not only affected by physiological factors, but also by the environment. No matter what kind of factors are affected, it will cause skin wrinkles, dryness, reduced elasticity, pigmentation and other problems. The appearance of these symptoms is due to the change of the collagen triple helix conformation of the facial skin, the decomposition of elastin polypeptides, and the disorder of the lipids filled in the skin. But these phenomena can be improved by controlling muscle contraction and supplementing collagen.

The anti-wrinkle products on the market mainly involve instant anti-wrinkle products and long-acting anti-wrinkle products. Instant anti-wrinkle focuses on blocking the conduction of neuromuscular current, so as to relax the facial muscles and instantly reduce wrinkles to achieve instant anti-wrinkle effect. Its products can meet people's temporary or short-term maintenance needs. Both CN110302072A and CN106038351B provide a fast wrinkle-removing composition, which can take effect quickly on the expression lines. Long-term anti-wrinkle is mainly achieved by promoting the synthesis of collagen and elastin, reducing the degradation of collagen and elastin, strengthening the basement membrane, and fighting against true wrinkles. Its products need long-term use to fundamentally reduce wrinkles. Compared with instant anti-wrinkle products, The effect is slower. The approaches of the above two products are relatively single, so the following technologies are available: CN112043664A, CN113171319B, CN115252456A, these technologies all provide a kind of instant and long-acting anti-wrinkle composition, wherein the composition provided by CN112043664A includes snake-like venom skin, bee venom skin, scorpion venom skin, cone snail skin, moon jellyfish extract, and anti-free radical components, but it is difficult to screen and compound such products, and the skins used are mainly from synthetic or fermentation sources, and the cost is relatively high , the price is higher after the product is made, and it is not matched with a penetration enhancer, which is likely to cause low bioavailability and poor effect; CN113171319B uses natural plant-derived ingredients to achieve instant and long-term wrinkle-removing effects, but it is effective Slow and requires long-term use.

Therefore, it is necessary to develop a product that simultaneously achieves instant and long-acting anti-wrinkle and anti-aging, has a good effect of promoting penetration, and has high bioavailability.

Contents of the invention

The purpose of the present invention is to provide a product that simultaneously achieves immediate and long-term anti-wrinkle and anti-aging effects, good permeation promoting effect and high bioavailability.

Technical scheme of the present invention is as follows:

An anti-wrinkle composition provided by the present invention, including: a polypeptide that induces the synthesis of elastin and type I collagen, an acetylcholine antagonist, a sodium ion channel inhibitor, a supramolecular ionic liquid, and HEPES (playing a synergistic role in promoting penetration) .

The anti-wrinkle composition provided by the present invention, said anti-wrinkle composition includes: a sodium ion channel inhibitor and an acetylcholine antagonist with an immediate anti-wrinkle effect, a polypeptide that induces the synthesis of elastin and type I collagen with a long-term anti-wrinkle effect, and a polypeptide with Supramolecular ionic liquids with permeation-enhancing effects. The anti-wrinkle composition provided by the present invention starts from the different action targets of the polypeptide, combines supramolecular ionic liquid to deliver the composite polypeptide, takes into account both specimens, and achieves the anti-wrinkle effect of 1+1>2. At the same time, supramolecular ionic liquids are also components of functional active substances in cosmetics. While promoting the absorption of active substances, they can also protect the structure of active substances and play a synergistic role.

Compared with the instant and long-acting anti-wrinkle composition in CN112043664A, the anti-wrinkle composition provided by the invention has the following technical advantages:

1. The introduction of a penetration enhancer can make active substances penetrate into the skin better;

2. Less peptides are used, and the types are simple, which reduces the burden on the skin;

3. The long-acting anti-wrinkle components in CN112043664A are snake-like peptides, melittin and Haiyue Shuimu extracts, but it is the snake-like peptides that actually work, and the role of melittin is anti-inflammatory. The function of the extract is moisturizing; and the long-acting anti-wrinkle component in the present invention is acetyl tetrapeptide-2.

Preferably, the polypeptide that induces the synthesis of elastin and type I collagen is acetyl tetrapeptide-2.

Preferably, the acetylcholine antagonist is one or both of snake venom and β-alanyl hydroxyprolyl diaminobutyryl benzylamide.

Preferably, the sodium ion channel inhibitor is conopeptide.

Preferably, the supramolecular ionic liquid is one or both of L-proline-lactic acid, L-proline-glycerol, and betaine-propylene glycol.

More preferably, the supramolecular ionic liquid is L-proline-lactic acid, L-proline-glycerol. L-proline/glycerol and L-proline/lactic acid constitute supramolecular ionic liquids, which can promote the penetration of compounded polypeptides; in addition, among all amino acids that form collagen, proline , which is often referred to as the "building block of protein". It is the main amino acid involved in collagen synthesis, accounting for 15% of the amino acid composition of collagen, and can be converted into hydroxylysine and hydroxyproline in the body to help form collagen. In addition, proline plays an important role in the structure of collagen. If the collagen lacks proline and cannot be hydroxylated, it will affect the stability of the collagen triple helix structure, thereby affecting the synthesis of the overall collagen.

Preferably, the anti-wrinkle composition further includes polyhydric alcohol, and the polyhydric alcohol is one or both of glycerin and 1,2-pentanediol. The polyols can act as preservatives.

Preferably, in parts by weight, the anti-wrinkle composition includes: 0.1-4.5 parts of acetyl tetrapeptide-2, 0.01-4 parts of snake venom peptide, 0.01-5 parts of conus peptide, 0.5-40 parts of supramolecular ionic liquid , 1-10 parts of HEPES, 1-62 parts of glycerin, 1-20 parts of 1,2-pentanediol.

In order to better understand the present invention, a detailed description will be given below in conjunction with a specific mechanism.

Conopeptide can block the conduction of neuromuscular current and instantly fade wrinkles. It can specifically block voltage-gated Na ⁺ channels, especially for muscle channel Na V1.4, and the blocking rate can reach more than 90%. It can block and maintain a certain signal flow, inhibit muscle contraction, but not tense and stiff, so as to achieve instant wrinkle removal effect. In addition, conopeptides help stimulate the synthesis and accumulation of extracellular matrix structural components, and can also help inhibit disordered collagen cross-linking and the release of metalloproteinase 1 (MMP-1); promote fibroblast regeneration, Stimulates the synthesis of collagen and restores skin elasticity.

Acetyl tetrapeptide-2 promotes the synthesis and enhances the activity of FBLN5 and LOXL1, thereby increasing the amount of elastin synthesis. In addition, acetyl tetrapeptide-2 can up-regulate the expression of genes related to collagen synthesis and focal adhesions, and induce elasticity protein, type I collagen synthesis, thereby increasing skin firmness and rebuilding epidermal structure.

Snake venom peptide is a small molecular polypeptide that mimics the activity of snake venom Waglerin-1, which is 5 times more effective than botulinum in reducing dynamic wrinkles. It mainly acts on the prominent posterior mode and is a reversible antagonist of the muscle nicotinic acetylcholine receptor (nmAChR). It binds to the epsilon subunit of nmAChR to block the binding of acetylcholine to the receptor, eventually leading to receptor closure. In the closed state Sodium ions cannot be ingested, cannot be depolarized, nerve excitation transmission is blocked, and muscles relax, so wrinkles are smoothed, and facial expressions are not affected.

The present invention provides a kind of preparation method of anti-wrinkle composition as above, wherein, comprise steps:

Mix deionized water and supramolecular ionic liquid, and ultrasonically disperse, then add acetylcholine antagonists, sodium ion channel inhibitors, polypeptides that induce elastin and type I collagen synthesis, and continue ultrasonication to obtain solution 1;

After mixing HEPES and deionized water, add to the solution 1 to obtain the solution 2;

Lactic acid (for pH adjustment) was added to the solution 2, stirred, and sieved to obtain the anti-wrinkle composition.

Description of drawings

Figure 1 is a graph showing the results of cumulative penetration of conopeptides in the composition.

Figure 2 is a graph showing the results of the intradermal retention of conopeptides in the composition.

Figure 3 is a graph showing the results of cumulative penetration of snake venom in the composition.

Fig. 4 is a graph showing the results of intradermal retention of snake venom in the composition.

Figure 5 is a graph showing the results of cumulative penetration of acetyl tetrapeptide-2 in the composition.

Fig. 6 is a graph showing the results of intradermal retention of acetyl tetrapeptide-2 in the composition.

Figure 7 is a graph of the relative viability of SIRC cells.

Figure 8 is a graph of the relative survival rate of keratinocytes.

Figure 9 is a graph showing the relative survival rate of human dermal fibroblasts.

Figure 10 is a histogram of the relative area of collagen fibers.

Figure 11 is a histogram of the average optical density value of Collagen IV.

Figure 12 is a histogram of the average optical density value of Collagen VII.

Figure 13 is a histogram of the average optical density value of Collagen XVII.

Figure 14 is a histogram of the average optical density value of Laminin 5.

Fig. 15 is a graph showing the test results of COLIV and COLVII.

Figure 16 is a histogram of c-fos gene expression.

Detailed ways

The present invention provides an anti-wrinkle composition and its preparation method and application. In order to make the purpose, technical solution and effect of the present invention more clear and definite, the present invention will be further described in detail below. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

Example 1

The formula of the anti-wrinkle composition of the present embodiment sees the following table 1:

Table 1

Configuration method: Weigh 84.53 parts of deionized water, 5 parts of 1,2-pentanediol, and 10 parts of glycerin in a beaker according to the required parts by weight, ultrasonically disperse for 10 minutes, then add 0.304 parts of snake venom and 0.016 parts of taro Spiropeptide and 0.15 parts of acetyl tetrapeptide-2 were stirred to dissolve and become transparent, and the ultrasonic wave was continued for about 10 minutes. The obtained solution was passed through a 400-mesh filter cloth to obtain an anti-wrinkle composition.

Example 2

The formula of the anti-wrinkle composition of the present embodiment is shown in the following table 2:

Table 2

name content Deionized water 76.53 snake venom 0.304 Conopeptide 0.016 Acetyl Tetrapeptide-2 0.15 1,2-pentanediol 5 HEPES 8 glycerin 10

Configuration method: Weigh 10 parts of deionized water, 5 parts of 1,2-pentanediol, and 10 parts of glycerin in a beaker according to the required parts by weight, ultrasonically disperse for 10 minutes, then add 0.304 parts of snake venom and 0.016 parts of taro Spiropeptide, 0.15 parts of acetyl tetrapeptide-2, stir to dissolve transparently, continue to sonicate for about 10 minutes to obtain solution 1; weigh 8 parts of HEPES and 66.53 parts of deionized water according to the required parts by weight and pre-dissolve it, add the solution In 1, stir to dissolve and become transparent to obtain solution 2, which is passed through a 400-mesh filter cloth to obtain an anti-wrinkle composition.

Example 3

The formula of the anti-wrinkle composition of the present embodiment is shown in the following table 3:

table 3

name content Deionized water 73.53 snake venom 0.304 Conopeptide 0.016 Acetyl Tetrapeptide-2 0.15 1,2-pentanediol 5 HEPES 8 L-Proline-Lactic Acid 1 L-Proline-Glycerol 1 glycerin 10 lactic acid 1

Configuration method: Weigh 10 parts of deionized water, 1 part of L-proline-lactic acid, 1 part of L-proline-glycerin, 5 parts of 1,2-pentanediol, 10 parts Put glycerin in a beaker, ultrasonically disperse for 10 minutes, then add 0.304 parts of snake venom peptide, 0.016 parts of conopeptide, and 0.15 parts of acetyl tetrapeptide-2, stir to dissolve transparently, continue ultrasonication for about 10 minutes, and obtain solution 1; according to the required weight Number of parts Weigh 8 parts of HEPES and 63.53 parts of deionized water, after pre-dissolving, add to solution 1 to obtain solution 2; add 1 part of lactic acid to solution 2, stir to dissolve and make it transparent, and pass through a 400-mesh filter cloth to obtain an anti-wrinkle composition .

Example 4

The formula of the anti-wrinkle composition of the present embodiment is shown in the following table 4:

Table 4

name content Deionized water 71.53 snake venom 0.304 Conopeptide 0.016 Acetyl Tetrapeptide-2 0.15 1,2-pentanediol 5 HEPES 10 L-Proline-Lactic Acid 1 L-Proline-Glycerol 1 glycerin 10 lactic acid 1

Configuration method:

Weigh 10 parts of deionized water, 1 part of L-proline-lactic acid, 1 part of L-proline-glycerin, 5 parts of 1,2-pentanediol, and 10 parts of glycerin in the emulsified In the pot, ultrasonically disperse for 10 minutes, then add 0.304 parts of venom-like peptides, 0.016 parts of cono peptides, and 0.15 parts of acetyl tetrapeptide-2, stir to dissolve and become transparent, and continue ultrasonication for about 10 minutes to obtain solution 1; according to the required parts by weight 10 parts of HEPES and 61.53 parts of deionized water were weighed and pre-dissolved, then added to solution 1 to obtain solution 2; 1 part of lactic acid was added to solution 2, stirred to dissolve and become transparent, and passed through a 400-mesh filter cloth to obtain an anti-wrinkle composition.

Example 5

The formula of the anti-wrinkle composition of the present embodiment is shown in the following table 5:

table 5

name content Deionized water 56.18 snake venom 0.304 Conopeptide 0.016 Acetyl Tetrapeptide-2 0.15 1,2-pentanediol 5 HEPES 8 L-Proline-Lactic Acid 0.5 Betaine-Propylene Glycol 30

Configuration method: Weigh 10 parts of deionized water, 0.5 parts of L-proline-lactic acid, 30 parts of betaine-propylene glycol, and 5 parts of 1,2-pentanediol in a beaker according to the required parts by weight, and ultrasonically disperse After 10 minutes, add 0.304 parts of snake venom peptide, 0.016 parts of conus peptide, and 0.15 parts of acetyl tetrapeptide-2, stir to dissolve and make it transparent, and continue ultrasonication for about 10 minutes to obtain solution 1; weigh 8 parts of HEPES according to the required parts by weight 1. After 46.18 parts of deionized water are pre-dissolved, add to solution 1, stir to dissolve and make it transparent, and pass through a 400-mesh filter cloth to obtain an anti-wrinkle composition.

test case 1

In order to further demonstrate the penetration-enhancing effect, the above-mentioned 5 examples were subjected to a transdermal absorption test.

Using pigskin as a model, the Franz diffusion cell was used to conduct an in vitro skin penetration test to detect the cumulative transdermal and intradermal retention of conopeptide, acetyl tetrapeptide-2, and snake venom in the anti-wrinkle composition at various time points. This is used to evaluate the skin penetration performance of the composition with the penetration enhancing system.

It can be seen from Figure 1 and Figure 2 that:

Example 1 showed a downward trend after reaching the peak in the 23rd hour;

Example 2 shows a downward trend after reaching the peak in the 23rd hour;

Embodiment 3 shows a downward trend after reaching the peak in the 23rd hour;

Example 4 shows a downward trend after reaching the peak in the 23rd hour;

Example 5 shows a downward trend after reaching the peak in the 22nd hour;

It can be seen from the amount of intradermal retention: Example 1>Example 2>Example 4>Example 5>Example 3.

In summary, it can be seen from the penetration trend that the trends presented by the five examples are almost the same, and example 5 reaches the highest peak first; at the 24th hour, the cumulative permeation amount of conus peptide is example 5 > example 4>embodiment 3>embodiment 1>embodiment 2; Therefore, the penetration promoting effect of conopeptide in the composition is better in embodiment 5, followed by embodiment 4.

It can be seen from Figure 3 and Figure 4 that:

Embodiment 1 showed a downward trend after reaching a small peak in the 6th hour, and had an upward trend after reaching the lowest point in 22 hours;

Embodiment 2 showed a downward trend after reaching a small peak in the 6th hour, and had an upward trend after reaching the lowest point in 23 hours;

Embodiment 3 reached a small peak in the 2nd hour, after having reached the highest peak in 22 hours, dropped to the lowest point in 23 hours, and then showed an increasing trend;

Embodiment 4 reached a small peak in the 2nd hour, after reaching the highest peak in 22 hours, it was a flat trend;

Embodiment 5 reached a small peak in the 4th hour, and after reaching the highest peak in 23 hours, it showed a downward trend;

As can be seen from the intradermal retention: Example 1>Example 2>Example 4>Example 3>Example 5.

In summary, it can be seen from the penetration trend that the overall trend of Example 4 and Example 5 is relatively gentle, and the intradermal retention of Example 4 is higher than that of Example 5, so the snake-like venom in the composition The penetration-promoting effect of the peptide in Example 4 is better.

It can be seen from Figure 5 and Figure 6 that:

In embodiment 1, after reaching the peak in the 22nd hour, it was extremely fast and dropped to the lowest point, and then showed an upward trend in 23 hours;

Example 2 showed a downward trend after reaching the peak in the 22nd hour, and reached the lowest point in the 23rd hour and showed a gentle trend;

After reaching the peak in the 23rd hour in embodiment 3, it was a very rapid downward trend;

Example 4 showed a slow downward trend after reaching the peak in the 22nd hour;

As can be seen from the intradermal retention: Example 4>Example 3>Example 2>Example 1>Example 5.

In summary, it can be seen from the penetration trend that Example 4 and the overall trend are relatively gentle, and the intradermal retention of Example 4 is high, so the acetyl tetrapeptide-2 peptide in the composition is the same as in Example 4. The effect of promoting penetration is better.

In view of the sum of the above conclusions, the L-proline-lactic acid and L-proline-glycerol systems of Example 4 have the best penetration promoting effects.

L-proline/glycerol and L-proline/lactic acid constitute supramolecular ionic liquids, which can promote the penetration of compounded polypeptides; in addition, among all amino acids that form collagen, proline , which is often referred to as the "building block of protein". It is the main amino acid involved in collagen synthesis, accounting for 15% of the amino acid composition of collagen, and can be converted into hydroxylysine and hydroxyproline in the body to help form collagen. In addition, proline plays an important role in the structure of collagen. If the collagen lacks proline and cannot be hydroxylated, it will affect the stability of the collagen triple helix structure, thereby affecting the synthesis of the overall collagen.

test case 2

Select embodiment 4 to carry out safety test

1. In vitro SIRC cell model eye irritation detection

The stimulating effect caused by the direct action of the compound on the cornea is similar to the cytotoxic damage effect, and the degree of corneal damage can be reflected by detecting the cytotoxicity index. In this experiment, rabbit corneal epithelial cells (SIRC) cultured in vitro were exposed to the composition for a short time to simulate acute corneal irritation, and the eye damage caused by the composition was predicted by calculating the relative survival rate of the cells.

The results are shown in Figure 7, from which we can see that:

When the mass concentration of the sample is 5%, 0.5%, and 0.05%, the SIRC cell survival rate of the composition is all greater than 70, indicating that the composition has no or slight irritation.

2. Toxicity detection of keratinocytes and human dermal fibroblasts

The samples were tested for cytotoxicity on keratinocytes, and the MTT model was used for detection. The test results are shown in FIG. 8 .

The samples were tested for cytotoxicity on human dermal fibroblasts, and the MTT model was used for detection. The test results are shown in FIG. 9 .

From Figure 8-9 we can see:

Based on keratinocytes, if the composition is within the mass concentration range of 5.000%, the keratinocyte survival rate is above 90%, and the safe concentration range is within 5%, IC50=8.5802% (95%CI: 7.7693%-9.3910%);

Based on human dermal fibroblasts, the composition is within 0.156%, and the survival rate of human dermal fibroblasts is above 90%, so the safe mass concentration range is within 0.156%, and IC50=9.3821%.

3. Chicken embryo chorioallantoic membrane vessel experiment

The chicken chorioallantoic membrane (CAM) test is a commonly used method for evaluating the irritation of cosmetics in vitro. Drop a certain test substance on the CAM, observe the changes in the blood vessels of the CAM within a certain period of time, and evaluate the eye irritation of the test substance according to the degree of damage to the CAM.

The test results are shown in Table 6 below:

Table 6

Experimental results:

The composition has an RC50 of 47.26% (95% Cl: 35.16%-63.16%) to chicken fetal chorioallantoic membrane, and is non-irritating.

Test case 3

Select Example 4, based on the anti-wrinkle efficacy evaluation of isolated skin tissue

Taking Ex vivo isolated skin tissue cultured in vitro as the research object, the anti-wrinkle efficacy of the tested sample was evaluated by detecting the changes in the content of Collagen IV, Collagen VII, Collagen XVII, Laminin 5 and collagen fibers in the isolated skin tissue.

1. Collagen fiber test:

The test results are shown in Table 7 and Figure 10 below.

Table 7. Summary of Collagen Fiber Test Results

Experimental results:

Compared with the BC group, the content of collagen fibers in the NC group decreased significantly, indicating that the stimulation conditions of this test were effective.

Compared with the NC group, the collagen fiber content of the PC group increased significantly, indicating that the positive control of this test is effective.

Compared with the NC group, the collagen fiber content of the sample composition was significantly increased.

2. Collagen IV test:

The test results are shown in Table 8 and Figure 11 below.

Table 8. Summary of Collagen IV protein content test results

Experimental results:

Compared with the BC group, the Collagen IV protein content in the NC group decreased significantly, indicating that the stimulation conditions of this test were effective.

Compared with the NC group, the Collagen IV protein content in the PC group increased significantly, indicating that the positive control of this test is effective.

Compared with the NC group, the Collagen IV protein content of the composition increased significantly.

3. Collagen VII test:

The test results are shown in Table 9 and Figure 12 below.

Table 9. Summary of Collagen VII protein content test results

Experimental results:

Compared with the BC group, the Collagen VII protein content in the NC group decreased significantly, indicating that the stimulation conditions of this test were effective.

Compared with the NC group, the Collagen VII protein content in the PC group increased significantly, indicating that the positive control of this test is effective.

Compared with the NC group, the Collagen VII protein content of the composition increased significantly.

4. Collagen XVII test:

The test results are shown in Table 10 and Figure 13 below:

Table 10. Summary of Collagen XVII protein content test results

Experimental results:

Compared with the BC group, the Collagen XVII protein content in the NC group decreased significantly, indicating that the stimulation conditions of this test were effective.

Compared with the NC group, the Collagen XVII protein content in the PC group increased significantly, indicating that the positive control of this test is effective.

Compared with the NC group, the Collagen XVII protein content of the composition increased significantly.

5. Laminin 5 test:

The test results are shown in Table 11 and Figure 14 below:

Table 11. Summary of Laminin 5 test results

Experimental results:

Compared with the BC group, the Laminin 5 protein content in the NC group decreased significantly, indicating that the stimulation conditions of this test were effective.

Compared with the NC group, the Laminin 5 protein content in the PC group increased significantly, indicating that the positive control of this test is effective.

Compared with the NC group, the Laminin 5 protein content of the sample composition increased significantly.

Test case 4

Selecting Example 4, Evaluation of Anti-wrinkle Efficacy Based on Fibroblasts

The fibroblasts were stimulated by UVA, and the anti-wrinkle efficacy of the tested samples was evaluated by detecting the changes in the content of type I collagen and the content of CollagenIV, CollagenVII, Decorin, Biglycan, Smad3, Smad7, MMP1, MMP3 and genes.

1. ECM synthesis related gene test:

The fibroblasts were stimulated with UVA, and the fibroblasts were treated with RNAiso Plus to collect samples. According to the operation manual of the HAELISA kit, RNA extraction, reverse transcription and fluorescent quantitative PCR operations were carried out.

COLⅣ, COLⅦ test:

The results are shown in Figure 15, from Figure 15 we can see that:

Compared with the BC group, the expression of Collagen Ⅳ and Collagen Ⅶ genes in the NC group were significantly down-regulated, indicating that the test stimulation conditions were effective;

Compared with the NC group, the expressions of Collagen Ⅳ and Collagen Ⅶ genes in the PC group were significantly up-regulated, indicating that the positive control of this test was effective;

Compared with the NC group, the expressions of Collagen Ⅳ and Collagen Ⅶ genes of the composition were significantly up-regulated, indicating that it can increase the firmness of the skin and has anti-wrinkle effect.

2. ECM degradation related gene test:

The fibroblasts were stimulated with UVA, and the fibroblasts were treated with RNAiso Plus to collect samples. According to the operation manual of the HAELISA kit, RNA extraction, reverse transcription and fluorescent quantitative PCR operations were carried out.

c-fos test:

c-fos is one of the components of AP1 protein. AP1 is activator protein 1, which is a nuclear transcription factor. After being activated by phosphorylation, it enters the nucleus to regulate the transcription of matrix metalloproteinases, thereby playing a certain role in the collagen degradation process. effect.

The result is shown in Figure 16, from Figure 16 we can see that:

Compared with the BC group, the expression of c-fos gene in the NC group was significantly up-regulated, indicating that the stimulation conditions of this test were effective;

Compared with the NC group, the expression of c-fos gene in the PC group was significantly down-regulated, indicating that the positive control of this test is effective;

Compared with the NC group, the c-fos gene expression of the composition was significantly down-regulated, indicating that the composition can inhibit the degradation of collagen and has anti-wrinkle effect.

It should be understood that the application of the present invention is not limited to the above examples, and those skilled in the art can make improvements or changes according to the above descriptions, and all these improvements and changes should belong to the scope of protection of the appended claims of the present invention.

Claims (10)

1. An anti-wrinkle composition, characterized in that it comprises: polypeptides that induce elastin and type I collagen synthesis, acetylcholine antagonists, sodium ion channel inhibitors, supramolecular ionic liquids, and HEPES. 2. The anti-wrinkle composition according to claim 1, wherein the polypeptide that induces the synthesis of elastin and type I collagen is acetyl tetrapeptide-2; And/or, the acetylcholine antagonist is one or both of snake venom and β-alanyl hydroxyprolyl diaminobutyryl benzylamide. 3. The anti-wrinkle composition according to claim 1, wherein the sodium ion channel inhibitor is conopeptide. 4. anti-wrinkle composition according to claim 1, is characterized in that, described supramolecular ionic liquid is one or both in L-proline-lactic acid, L-proline-glycerol, betaine-propylene glycol kind. 5. The anti-wrinkle composition according to claim 1, characterized in that, the anti-wrinkle composition further comprises a polyhydric alcohol, and the polyhydric alcohol is one or both of glycerin and 1,2-pentanediol. 6. The anti-wrinkle composition according to claim 1, characterized in that, in parts by weight, the anti-wrinkle composition comprises: 0.1-4.5 parts of acetyl tetrapeptide-2, 0.01-4 parts of snake venom, 0.01-4 parts 5 parts of conus peptide, 0.5-40 parts of supramolecular ionic liquid, 1-10 parts of HEPES, 1-62 parts of glycerin, 1-20 parts of 1,2-pentanediol. 7. The anti-wrinkle composition according to claim 1, characterized in that, in parts by weight, the anti-wrinkle composition is made up of the following components: 0.15 part of acetyl tetrapeptide-2, 0.304 part of snake venom, 0.016 part of Cono peptide, 1 part L-proline-lactic acid, 1 part L-proline-glycerol, 10 parts HEPES, 10 parts glycerin, 5 parts 1,2-pentanediol, 71.53 parts deionized water, 1 part lactic acid. 8. A preparation method of the anti-wrinkle composition according to any one of claims 1-7, characterized in that, comprising the steps of: Mix deionized water and supramolecular ionic liquid, and ultrasonically disperse, then add acetylcholine antagonists, sodium ion channel inhibitors, polypeptides that induce elastin and type I collagen synthesis, and continue ultrasonication to obtain solution 1; After mixing HEPES and deionized water, add to the solution 1 to obtain the solution 2; Add lactic acid into the solution 2, stir and sieve to obtain the anti-wrinkle composition. 9. the preparation method of anti-wrinkle composition according to claim 8, is characterized in that, the preparation method of described anti-wrinkle composition specifically comprises the steps: Take 10 parts of deionized water, 1 part of L-proline-lactic acid, 1 part of L-proline-glycerol, 5 parts of 1,2-pentanediol, and 10 parts of glycerin in an emulsifying pot, ultrasonically disperse for 10 minutes, and then Add 0.304 parts of venom-like peptides, 0.016 parts of conopeptides, and 0.15 parts of acetyl tetrapeptide-2, and continue to sonicate for 10 minutes to obtain solution 1; After mixing 10 parts of HEPES and 61.53 parts of deionized water, add it to solution 1 to obtain solution 2; Add 1 part of lactic acid into solution 2, stir until dissolved and transparent, and pass through a 400-mesh filter cloth to obtain the anti-wrinkle composition. 10. The application of the anti-wrinkle composition described in any one of claims 1-7 in the preparation of cosmetics.