CN118161435B

CN118161435B - Wrinkle removing and resisting composition containing coastal cress tissue culture

Info

Publication number: CN118161435B
Application number: CN202410208886.2A
Authority: CN
Inventors: 陈影霞
Original assignee: Huaze Ruifu Biotechnology Guangzhou Co ltd
Current assignee: Huaze Ruifu Biotechnology Guangzhou Co ltd
Priority date: 2024-02-26
Filing date: 2024-02-26
Publication date: 2024-08-23
Anticipated expiration: 2044-02-26
Also published as: CN118161435A

Abstract

The invention belongs to the field of cosmetic preparations, and particularly relates to a wrinkle removing and resisting composition containing a coastal cress tissue culture. The composition comprises the following components: coastal cress callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, tocopherol, dipotassium glycyrrhizinate; the method also comprises the following steps: flos Althaeae Roseae extract and herba Cypress branch extract. The wrinkle-removing and wrinkle-removing composition disclosed by the invention has certain effects of tightening skin, removing wrinkles and resisting wrinkles, and has good synergistic effect by compounding the littoral cress callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, tocopherol (vitamin E) and dipotassium glycyrrhizinate with plant extract composition hollyhock flower extract and water cypress branch extract.

Description

Wrinkle removing and resisting composition containing coastal cress tissue culture

Technical Field

The invention belongs to the field of cosmetic preparations, and particularly relates to a wrinkle removing and resisting composition containing a coastal cress tissue culture.

Background

Callus refers to tissue which is newly grown on the surface of a wound after the local part of the plant body is stimulated by the wound. Generally refers to tissue in plants that consists of parenchyma cells and does not form a specific structure, and is usually found at the wound of the plant. Modern researches have found that plant callus contains various active functional components, and has been well accepted and applied in the fields of medicine, cosmetics and the like because of good safety and reliability. For example, the tobacco callus extract has the effects of resisting acne and treating glaucoma, the saussurea involucrata callus extract has the effects of resisting tumor, the xanthium sibiricum callus extract has the effects of inhibiting bacteria, and the rhodiola sachalinensis callus has the effects of resisting oxidation.

However, the research degree of the callus extract is limited, and no report on the synergistic effect of the callus extract combined with other components exists. Accordingly, it is desirable to provide a composition containing coastal cress callus culture filtrate that is excellent in performance.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a composition containing coastal cress callus culture filtrate and application thereof. The composition generates a certain synergistic effect through the cooperation of the plant callus extract and other components, and achieves remarkable effects of removing wrinkles, resisting oxidization and aging.

In order to achieve the above purpose, the present invention discloses the following technical solutions:

In a first aspect, the invention provides a composition comprising the following components in parts by weight:

Further, the composition comprises the following components in parts by weight:

still further, the composition also comprises: flos Althaeae Roseae extract and herba Cypress branch extract.

Further preferably, the hollyhock flower extract is 0.12-0.15 parts by mass; the water cypress branch extract is 0.5-1 part.

In a second aspect, the present invention provides a wrinkle-reducing, anti-wrinkle skin-resident formulation comprising the composition of the first aspect.

Further, the composition of the first aspect is added to the wrinkle-removing and anti-wrinkle skin resident agent in an amount of 0.17% -1.7% by mass.

Further, the skin resident agent for removing the wrinkles and resisting the wrinkles further comprises an emulsifying agent, a preservative, a humectant, a pH regulator and a solvent.

Further, the emulsifier is at least one of acrylic acid (ester)/C10-30 alkanol acrylate cross-linked polymer, C14-22 alcohol and C12-20 alkyl glucoside.

Further, the humectant is at least one of glycerin, sodium hyaluronate, squalane, butanediol and 1, 2-hexanediol;

the preservative is at least one of p-hydroxyacetophenone, phenoxyethanol and ethylhexyl glycerol;

The pH regulator is at least one of arginine, sodium hydroxide and disodium hydrogen phosphate.

In a third aspect, the present invention provides a method for preparing the wrinkle-removing and anti-wrinkle skin resident agent according to the second aspect, comprising the following steps:

(1) Heating a certain amount of preservative, humectant, emulsifier and solvent to 65-85deg.C for dissolving, stirring, homogenizing and mixing, and maintaining the temperature for 20min or more to obtain a mixture;

(2) Cooling the mixture obtained in the step (1) to 40-50 ℃, adding the composition according to any one of claims 1-4 into the mixture, dissolving, stirring, mixing, adding a pH regulator, and regulating the pH value to obtain the skin residence agent for removing wrinkles.

In the invention, the following components are added:

The coastal cress callus culture filtrate can effectively promote the natural generation of skin collagen, elastic fiber and other active proteins.

Palmitoyl tripeptide-1 can promote the synthesis of extracellular cytoplasm such as type I and III collagen, elastin and structural glycoprotein such as laminin and fibronectin, promote wound restoration, promote the upgrading and reconstruction of the surface layer of skin, make the skin more compact and elastic, and simultaneously facilitate the repair of the skin after ultraviolet irradiation.

The palmitoyl tetrapeptide-7, palmitoyl tripeptide-1 and acetyl hexapeptide-8 are used in a compound way, so that the synergistic effect is achieved, the generation of interleukin IL-6 in keratinocytes and fibroblasts caused by external injury and ultraviolet is effectively reduced, inflammatory reaction is reduced, skin elasticity is protected and stable, and skin aging is delayed.

Acetyl hexapeptide-8, also called argireline, botulinum-like, etc., is a small molecule protein composed of six amino acids, and has the effects of mainly removing wrinkles, promoting collagen production, repairing damaged skin, and improving cellular metabolism.

The tocopherol is an important phenolic antioxidant, can prevent lipid peroxidation, protect the integrity of cell membranes, reduce the damage of ultraviolet rays to skin, has anti-aging effect, and can promote metabolism of skin and improve moisture and elasticity of skin.

The main chemical components in the hollyhock flower extract comprise flavone and flavonoid glycoside compounds, wherein the flavonoid glycoside compounds comprise quercetin, hyperoside, myricetin-3 '-O-beta-D-glucoside, quercetin-3' -glucoside and the like, and the flavonoid glycoside compounds are very excellent antioxidant components, have higher antioxidant capacity than vitamin C and vitamin E, and can help to remove free radicals and delay aging.

The extract of herba Cypress has positive effects of balancing skin oil secretion, relieving acne, etc., and enhancing transdermal absorption rate. After the water cypress branch extract and the hollyhock flower extract are compounded and used, the wrinkle removing and resisting effects of skin on the composition coastal cress callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7 and acetyl hexapeptide-8 can be improved under the condition of the same dosage.

The invention has the beneficial effects that:

1. The wrinkle removing and resisting composition containing the littoral celery callus culture filtrate provided by the invention has good effects of tightening skin, removing wrinkles and resisting wrinkles;

2. The coastal thorny celery callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, tocopherol (vitamin E) and dipotassium glycyrrhizinate are compounded with the plant extract composition hollyhock flower extract and the water cypress branch extract, so that the invention has good synergistic effect, and the wrinkle removing and resisting effects of the skin resident agent formula are improved.

Detailed Description

For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the following specific examples. It will be appreciated by persons skilled in the art that the specific embodiments described herein are for purposes of illustration only and are not intended to be limiting.

The test methods used in the specific embodiments are all conventional methods unless otherwise specified; materials, reagents, and the like used, unless otherwise specified, are commercially available; the percentages mentioned in the detailed description are mass percentages unless otherwise indicated.

In the invention, the following components are added:

The coastal cress callus culture filtrate was coastal cress (ERYNGIUM MARITIMUM) callus culture filtrate, INCI name ERYNGIUM MARITIMUM CALLUS CULTURE FILTRATE, available from SEPPIC, france.

The sea fennel callus culture filtrate was sea fennel (CRITHMUM MARITIMUM) callus culture filtrate, INCI name CRITHMUM MARITIMUM CALLUS CULTURE FILTRATE, available from SEPPIC, france.

The EXTRACT of herba Cypress is EXTRACT of herba Cypress (MYRICARIA GERMANICA), INCI named MYRICARIA GERMANICA EXTRACT, available from Shaanxi Sihai Biotechnology Co.

The flos Althaeae Roseae extract is flos Althaeae Roseae (ALTHAEAROSEA) extract, INCI name ALTHAEA ROSEAFLOWER EXTRACT, available from Shaanxi Sisea Biotechnology Co.

Other raw materials are available commercially.

Preparation of wrinkle-removing and anti-wrinkle composition

The raw materials are weighed according to the mass parts in the table 1, and then stirred and mixed uniformly to obtain the anti-wrinkle composition.

Table 1 raw materials for preparing the anti-wrinkle composition

Note that: the raw materials in the table are in parts by mass, "-" means not added.

Preparation of skin resident agent containing anti-wrinkle composition

Step (1), taking acrylic acid (ester)/C10-30 alkanol acrylate crosslinked polymer, glycerin, sodium hyaluronate, dipotassium glycyrrhizinate and water according to the mass parts of table 2, stirring and heating to 85 ℃, preserving heat for 20 minutes, and stirring and dissolving completely to obtain a mixture a;

step (2), taking C14-22 alcohol, C12-20 alkyl glucoside, squalane and tocopherol (vitamin E), heating to 80 ℃, stirring and dissolving completely, and preserving heat for 10 minutes to obtain a mixture b;

Step (3), adding the mixture a and the mixture b into an emulsifying pot, stirring at 80 ℃, homogenizing until complete emulsification is uniform, and obtaining a mixture c;

step (4), cooling the mixture c to 65 ℃, adding butanediol, 1, 2-hexanediol and p-hydroxyacetophenone, and stirring until the mixture is completely and uniformly dispersed to obtain a mixture d;

Step (5), cooling the mixture d to 45 ℃, sequentially adding palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, coastal thorn celery callus culture filtrate, hollyhock flower extract and water cypress branch extract, and stirring until the mixture is completely and uniformly mixed to obtain a mixture e;

And (6) cooling the mixture e to 38 ℃, adding arginine dissolved in water in advance, and regulating the pH value to be more than or equal to 6.5 to obtain the skin resident agent for removing the wrinkles.

TABLE 2 raw materials for skin resident agent

Performance test:

inhibition of elastase assay

The test uses porcine pancreatic elastase as a test subject and N-succinyl-alanine-p-nitroaniline (AAAPVN) as a substrate. The porcine pancreatic elastase can hydrolyze AAAPVN, and the hydrolysis product can cause the increase of absorbance at 420nm wavelength, and the absorbance is measured by an enzyme-labeled instrument, so that the tightening and anti-wrinkle effects of the sample to be measured can be evaluated.

1. Solution preparation

(1) Sample to be measured: the above prepared wrinkle-removing and wrinkle-resisting compositions 1-7 are respectively taken.

(2) Tris-HCl buffer (0.1M PH=8.0) was prepared: 2.42gTris was weighed into a beaker, 200mL of ultrapure water was added, and after dissolution was completed, the pH was adjusted to 8.0 with concentrated HCl.

(3) Configuration of positive control tea polyphenol solution (1 mg/ml): 5mg of tea polyphenols were weighed and dissolved in 5ml of Tris-HCl buffer.

(4) Substrate solution AAAPVN (2 mM) was prepared: 4.51mg of N-succinyl-alanine-p-nitroaniline are weighed out and dissolved in 5ml of Tris-HCl buffer.

(5) Porcine pancreatic elastase solution (0.171U/mL) was prepared: mu.l of the solution was dissolved in 10ml of Tris-HCl buffer.

2. Grouping and sampling

The test is divided into 4 groups, namely a sample group to be tested, a positive control group, a blank control group and a model control group, and 4 compound holes are arranged under the same concentration of the same group. The amounts of each solution added are shown in Table 3.

Table 3 test group

3. Measurement method

The reaction was allowed to stand at room temperature for 15min, and the absorbance was measured at 420nm with a microplate reader.

4. Calculation formula

Wherein: average value of absorbance of a ₀ -blank wells;

A ₁ -average value of absorbance of sample wells;

Average value of absorbance of a ₂ -model control wells.

5. Statistical method

Statistical analysis was performed using the SPSS22.0 statistical software package to descriptive statistics of the measurements of the test area. The change in the analytical value was calculated and the difference between the control group and the sample group. If the test data are normally distributed, adopting an independent T test method to carry out statistical analysis; if the test data are in non-normal distribution, adopting a rank sum test method to carry out statistical analysis. The statistical methods all use a two-tailed test with a test level α=0.05.

6. Experimental results

TABLE 4 results of porcine pancreatic elastase inhibition rate

Promote type I collagen synthesis assay

In the test, fibroblasts are taken as a detection object, a cell photoaging damage model is established through UVA irradiation, the change of the content of type I Collagen (Collagen I) after the action of a sample to be detected is detected, and the tightening and anti-wrinkle effects of the sample to be detected are evaluated.

(1) Inoculating: cells were seeded at a seeding density of 1×10 ⁴ cells/well into 96-well plates and incubated overnight in an incubator (37 ℃, 5% co ₂).

(2) UVA irradiation: after washing the cells with PBS, the groups with UVA irradiation were subjected to UVA irradiation of 30J/cm ² according to the experimental group of Table 5.

(3) Preparing liquid: compositions 1-7 were selected as test sample solutions, respectively, and experiments were performed using TGF-beta (concentration 100 ng/mL) as a positive control. 100 μl was loaded per well, and 3 duplicate wells were set per group. After the completion of the administration, the 96-well plate was placed in an incubator at 37℃and incubated for 24 hours in the dark.

TABLE 5 grouping of experiments

(4) Cell supernatant collection: taking out the 96-well plate 24h after administration, respectively collecting cell supernatant corresponding to each sample group into a centrifuge tube, centrifuging at 1000rpm for 10min, collecting supernatant, placing the supernatant into a 1.5mL centrifuge tube, and storing at-20 ℃ for later use.

(5) ELISA kit for determining type I collagen concentration in cell supernatant: taking out the kit and the sample to be tested 30min before testing, and placing the kit and the sample to be tested at room temperature for use, wherein the testing operation is strictly operated according to the instruction of the kit.

TABLE 6 type I collagen content test results

Antioxidant test (DPPH radical scavenging)

(1) Test design

Preparing DPPH into 2X 10 ^-4 mol/L solution with absolute ethyl alcohol, preparing 2mL of each of the solution to be detected, the absolute ethyl alcohol and the DPPH solution as 2-7 parts by taking the composition as the solution to be detected, respectively uniformly mixing, standing at room temperature for 30 minutes, measuring absorbance at a wavelength of 517nm to obtain Ai, measuring each sample in parallel for 3 times, and taking an average value.

Vc is used as a positive control, absolute ethyl alcohol is used for replacing DPPH to measure absorbance value to be used as a control Aj blank, and DPPH solution and absolute ethyl alcohol are mixed to measure absorbance Ac.

The clearance of DPPH radicals from each sample to be tested was calculated according to the following formula (1) and recorded in the following table 7.

Clearance (%) = (1- (Ai-Aj)/Ac) ×100%........... (1)

Wherein: ai is absorbance of 2mL of the mixed solution of the liquid to be detected and the reagent of 2 mLDPPH; aj is absorbance of 2mL of to-be-detected liquid and 2mL of absolute ethyl alcohol mixed liquid; ac is the absorbance of a 2mLDPPH solution+2 mL absolute ethanol mixture.

TABLE 7DPPH clearance rate

Group of	DPPH clearance rate
		Composition 1	73.14％
Composition 2	76.41％
		Composition 3	72.37％
Composition 4	43.63％
		Composition 5	66.34％
Composition 6	63.71％
		Composition 7	32.11％

Analysis of results

1. According to the test result, the anti-wrinkle composition has certain effects of tightening skin and removing wrinkles;

2. According to the test results, the coastal thorny celery callus culture filtrate, the palmitoyl tripeptide-1, the palmitoyl tetrapeptide-7, the acetyl hexapeptide-8, the tocopherol (vitamin E) and the dipotassium glycyrrhizinate in the anti-wrinkle composition have the effect of promoting skin repair together with the hollyhock flower extract and the water cypress branch extract which are plant extract compositions, and under the synergistic effect, the capability of inhibiting the activity of elastase can be effectively improved, the synthesis of skin collagen can be promoted, and the scavenging effect on free radical ions can be enhanced;

3. From the test results, it is clear that the addition amounts of the functional components (coastal cress callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, tocopherol (vitamin E), dipotassium glycyrrhizinate, hollyhock flower extract and water cypress branch extract) of the composition 2 in the compositions 1, 2 and 3 have the optimal performance compared with other compositions.

Skin resident agent examples 1-4 efficacy test

Stability test:

1. Heat resistance test: the temperature of the constant temperature incubator is regulated to 40 ℃, the prepared examples are respectively used as 4 groups of samples, three samples of each group are taken and are filled in transparent glass bottles, the sample filling amount is 20 ml/bottle, the samples are placed in the constant temperature incubator after being sealed, and the samples are taken out after three months, are restored to room temperature, and are observed for appearance change.

2. Cold resistance test: and (3) regulating the temperature of the constant temperature incubator to-10 ℃, taking three samples from each group, filling the three samples into transparent glass bottles, sealing the bottles with the sample filling quantity of 20 ml/bottle, putting the bottles into the constant temperature incubator for three months, taking out the bottles, recovering the bottles to room temperature, and observing the appearance change.

3. And (3) normal temperature test: and (3) putting the 4 groups of samples into transparent glass bottles, wherein the sample loading amount is 20 ml/bottle, sealing, standing for 6 months at normal temperature, and observing the appearance change of the emulsion.

No phenomena such as layering, precipitation, color change and the like are observed in the heat resistance test, the cold resistance test and the normal temperature test, and the original appearance is maintained.

Human skin patch test:

Examples 1-4 of the foregoing preparations were referred to the human skin patch test in 2022 cosmetic safety Specification.

Skin reactions were observed as standard at 30min (after the disappearance of the indentations), 24h and 48h, respectively, and the observations were recorded, see table 5.

TABLE 8 human safety test results

Numbering device	30min	24h	48h
				Example 1	Grade 0, 30	Grade 0, 30	Grade 0, 30
Example 2	Grade 0, 30	Grade 0, 30	Grade 0, 30
				Example 3	Grade 0, 30	Grade 0, 30	Grade 0, 30
Example 4	Grade 0, 30	Grade 0, 30	Grade 0, 30

Subjective evaluation test of cosmetic effect:

a subject: 40 healthy volunteers, sex women, age 45.+ -.2 years, were selected and divided into 4 groups (i.e., skin resident agents prepared in examples 1-4).

The using method comprises the following steps: the prepared liquid cosmetic was applied to the face in the morning and evening every day for 4 weeks. During the test, the volunteer could not change the original facial cleansing pattern and habit. In addition, other cosmetics than the test products could not be used.

The evaluation method comprises the following steps: the subjective evaluation method was used to evaluate the change in facial wrinkles, no change was noted as×, slight change was noted as ①, the reduction in wrinkles was more than ②, and the reduction in wrinkles was noted as ③, and the results are shown in table 9.

Table 9 subjective evaluation results

Analysis of results:

According to experimental results, compared with examples 2-4, the embodiment 1 has obvious wrinkle removing effect, which shows that the composition provided by the invention has good synergistic effect with the compounding of plant extract composition hollyhock flower extract and water cypress branch extract, namely the littoral root callus culture filtrate, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, tocopherol (vitamin E) and dipotassium glycyrrhizinate, and the wrinkle removing and resisting effect of the whole formula is improved.

Finally, it should be noted that the above embodiments and comparative examples are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. The composition is characterized by comprising the following components in parts by mass:

0.02-0.08 part of coastal cress callus culture filtrate;

0.0002 to 0.0004 parts of palmitoyl tripeptide-1;

0.00010 to 0.00020 parts of palmitoyl tetrapeptide-7;

0.002-0.006 parts of acetyl hexapeptide-8;

0.1-0.3 parts of tocopherol;

0.05-0.15 part of dipotassium glycyrrhizinate;

0.12-0.15 part of hollyhock flower extract;

0.5-1 part of water cypress branch extract.

2. The composition according to claim 1, characterized by comprising the following components in parts by mass:

0.05 part of coastal cress callus culture filtrate;

palmitoyl tripeptide-1.0003 parts;

0.00015 parts of palmitoyl tetrapeptide-7;

0.004 part of acetyl hexapeptide-8;

0.2 parts of tocopherol;

0.1 part of dipotassium glycyrrhizinate;

0.12-0.15 part of hollyhock flower extract;

0.5-1 part of water cypress branch extract.

3. A wrinkle-removing and anti-wrinkle skin-resident agent comprising the composition according to claim 1 or 2.

4. The wrinkle-removing and anti-wrinkle skin resident agent according to claim 3, wherein the composition according to claim 1 or 2 is added to the wrinkle-removing and anti-wrinkle skin resident agent in an amount of 0.17% to 1.7% by mass.

5. The wrinkle-removing and anti-wrinkle skin resident agent according to claim 3, further comprising an emulsifier, a preservative, a humectant, a pH adjustor, and a solvent.

6. The anti-wrinkle skin resident agent according to claim 5, wherein the emulsifier is at least one of acrylic acid (esters)/C10-30 alkanol acrylate cross-linked polymer, C14-22 alcohol, and C12-20 alkyl glucoside.

7. The anti-wrinkle skin resident agent according to claim 5, wherein the humectant is at least one of glycerin, sodium hyaluronate, squalane, butylene glycol, 1, 2-hexanediol;

8. The method for preparing the wrinkle-removing and wrinkle-preventing skin resident agent according to any one of claims 3 to 7, comprising the steps of:

(2) Cooling the mixture obtained in the step (1) to 40-50 ℃, adding the composition according to the claim 1 or 2 into the mixture, dissolving, stirring, mixing, adding a pH regulator, and regulating the pH value to obtain the skin residence agent for removing wrinkles and resisting wrinkles.