CN110934774A - Wrinkle-removing and tightening stock solution containing recombinant collagen - Google Patents

Abstract

The invention discloses a wrinkle-removing and tightening stock solution containing recombinant collagen, belonging to the field of cosmetics. The wrinkle-removing and tightening stock solution comprises the following components in percentage by mass: 0.1-1% of recombinant collagen, 10-40% of butanediol, 1-5% of 1, 2-hexanediol, 81-10% of polyethylene glycol, 0.1-0.5% of sodium hyaluronate, 0.1-0.5% of polyglutamic acid, 0. 80.05-2% of acetyl hexapeptide and the balance of water. The invention adopts the recombinant collagen from microbial fermentation to be compounded with other raw materials, and the problems of precipitation, color change, precipitation and the like can not occur. The wrinkle-removing and tightening stock solution disclosed by the invention has an excellent moisturizing and water-locking effect on skin and has the effects of tightening and removing wrinkles.

Description

Wrinkle-removing and tightening stock solution containing recombinant collagen

Technical Field

The invention belongs to the field of cosmetic preparations, and relates to a wrinkle-removing and tightening stock solution containing recombinant collagen.

Background

Women begin to lose collagen at 20 years of age, enter peak periods of loss at 25 years of age, and have collagen content less than half of 18 years of age at 40 years of age. Loss of collagen means the appearance of wrinkles and aging of the appearance. Anti-aging cosmetic formulations are receiving increasing attention and are continuously increasing in total volume on the market each year. Sufficient collagen is supplemented to achieve the effects of tightening skin and removing wrinkles. At present, most of the collagen used in wrinkle-removing products on the market is extracted from animal bones or skin, for example, marine collagen extracted from deep-sea fish. However, because the collagen is derived from animals, the potential biological safety hazard still exists, the wide application of the collagen in various products such as cosmetics, foods, medicines and the like is limited, and the wrinkle-removing and tightening effects of the collagen are still not ideal.

Disclosure of Invention

The invention provides a wrinkle-removing and tightening stock solution containing recombinant collagen. The wrinkle-removing firming stock solution takes recombinant collagen with excellent skin affinity and histocompatibility as a raw material, increases the moisture-preserving and water-locking capacity of the skin, and has good wrinkle-removing effect.

The technical scheme of the invention is as follows:

the anti-wrinkle firming stock solution containing the recombinant collagen comprises the following components in percentage by mass: 0.1-1% of recombinant collagen, 10-40% of butanediol, 1-5% of 1, 2-hexanediol, 81-10% of polyethylene glycol, 0.1-0.5% of sodium hyaluronate, 0.1-0.5% of polyglutamic acid, 0. 80.05-2% of acetyl hexapeptide and the balance of water.

Preferably, the wrinkle-removing and tightening stock solution comprises the following components in percentage by mass: 0.4% of recombinant collagen, 30% of butanediol, 2.5% of 1, 2-hexanediol, 0.4% of polyethylene glycol-810%, 0.4% of sodium hyaluronate, 0.4% of polyglutamic acid, 0-80.07% of acetyl hexapeptide and the balance of water.

In the wrinkle-removing and firming stock solution, butanediol is used as a humectant, 1, 2-hexanediol is used as a humectant, polyethylene glycol-8 is used as a humectant, sodium hyaluronate is used as a humectant, polyglutamic acid is used as a humectant, acetyl hexapeptide-8 is used as a skin conditioner, and recombinant collagen is used as the skin conditioner.

As a preferable scheme of the invention, the recombinant collagen is produced by fermenting Pichia pastoris with the preservation number of CGMCCNo.5021. The recombinant collagen adopted by the invention takes a human collagen gene as a template, is optimized and modified, is introduced into pichia pastoris, and is obtained by fermentation induction, separation and purification. The recombinant collagen is highly consistent with human collagen, and has good skin affinity and tissue compatibility.

The invention also provides a preparation method of the wrinkle-removing and tightening stock solution containing the recombinant collagen, which comprises the following steps:

adding sodium hyaluronate, polyglutamic acid and recombinant collagen into butanediol according to a certain proportion, stirring until uniformly mixing, then adding 1, 2-hexanediol, polyethylene glycol-8 and acetyl hexapeptide-8, adding water to 100%, stirring until completely dissolving, and obtaining the wrinkle-removing and tightening stock solution containing the recombinant collagen.

Compared with the prior art, the invention has the following advantages:

the invention adopts the molecular weight determination of the recombinant collagen from microbial fermentation, is the single-component collagen and has no peculiar smell. It has good water solubility, maximum solubility of 20%, good transdermal property, and easy absorption. In addition, the recombinant collagen is derived from microbial fermentation, has good performance when being compounded with other raw materials, and can not generate the problems of precipitation, color change, precipitation and the like. The wrinkle-removing tightening stock solution disclosed by the invention is prepared by compounding the recombinant collagen with sodium hyaluronate, polyglutamic acid and the like, so that the wrinkle-removing tightening stock solution not only has an excellent moisturizing and water-locking effect on skin, but also has the effects of tightening and removing wrinkles.

Drawings

Fig. 1 is a graph showing wrinkle improvement rates in T, B and C groups after 8 weeks of use.

Fig. 2 is a graph of pore improvement rates in groups T, B, and C after 2 weeks of use.

Fig. 3 is a graph of pore improvement after 8 weeks of use in groups T, B and C.

Fig. 4 is a graph showing the improvement rate of moisture in T, B and C groups after 8 weeks of use.

Fig. 5 is a graph of texture improvement using groups T, B and C after 8 weeks.

Fig. 6 is a real object diagram (a) of the left canthus and a real object diagram (B) of the right canthus of subject 1 in group C before use (W0) and after use for 8 weeks (W8).

Detailed Description

The present invention will be described in more detail with reference to the following examples and drawings, which are given by way of illustration only and are not intended to limit the scope of the present invention.

Example 1

The anti-wrinkle firming stock solution containing the recombinant collagen comprises the following components in percentage by mass: 0.4% of recombinant collagen, 30% of butanediol, 2.5% of 1, 2-hexanediol, 0.4% of polyethylene glycol-810%, 0.4% of sodium hyaluronate, 0.4% of polyglutamic acid, 0-80.07% of acetyl hexapeptide and the balance of water. Is prepared by the following steps: according to the formula proportion, adding sodium hyaluronate, polyglutamic acid and recombinant collagen into butanediol, stirring until the mixture is uniformly mixed, then adding 1, 2-hexanediol, polyethylene glycol-8 and acetyl hexapeptide-8, adding water to 100%, and uniformly stirring until the mixture is completely dissolved to obtain the wrinkle-removing and tightening stock solution containing the recombinant collagen.

In the embodiment, the recombinant collagen is produced by fermenting Pichia pastoris with the preservation number of CGMCC No.5021, and other raw materials are conventional products sold in the market.

In this embodiment, the raw materials may be flexibly combined within a given mixture ratio range, which is not always enumerated herein.

Example 2

1. Experimental products:

and (3) frosting: 68.65% of water, 0.2% of carbomer, 0.2% of triethanolamine, 0.1% of xanthan gum, 4% of glycerol, 3% of butanediol, 20.5% of bioglycan gum, 1% of cetostearyl alcohol, 0.8% of C20-22 alcohol phosphate (and) C20-22 alcohol, 0.5% of glyceryl stearate, 3% of caprylic/capric glycerides, 3% of polydimethylsiloxane, 4.5% of shea butter (Butyrrhiza PARKII), 0.15% of methylparaben, 0.4% of caprylyl hydroxamic acid and 0.5% of 1, 2-hexanediol.

The wrinkle-removing and tightening stock solution containing the recombinant collagen comprises the following components in percentage by mass: 0.4% of recombinant collagen, 30% of butanediol, 2.5% of 1, 2-hexanediol, 0.4% of polyethylene glycol-810%, 0.4% of sodium hyaluronate, 0.4% of polyglutamic acid, 0-80.07% of acetyl hexapeptide and the balance of water.

The wrinkle-removing and tightening stock solution without recombinant collagen comprises the following components in percentage by mass: 30% of butanediol, 2.5% of 1, 2-hexanediol, 0.4% of polyethylene glycol-810%, 0.4% of sodium hyaluronate, 0.4% of polyglutamic acid, 0-80.07% of acetyl hexapeptide and the balance of water.

2. Experiment grouping

The experiment is divided into a group T, a group B and a group C, the same batch of cream with the same formula is used as a substrate, the authenticity of the experiment is ensured, and the group T is a placebo group, and 10% purified water is added for homogenizing and stirring uniformly; group B is a control group, and 10% of wrinkle-removing and tightening stock solution without recombinant collagen is added, homogenized and stirred uniformly; and C, adding 10% of wrinkle-removing and tightening stock solution containing recombinant collagen, homogenizing and stirring uniformly.

Group T (placebo group): frost + 10% purified water;

group B (control group): cream + 10% wrinkle-removing tightening stock solution without recombinant collagen;

group C (experimental group): cream + 10% wrinkle-removing firming stock solution containing recombinant collagen.

3. The test purpose is as follows: open 8 week test, changes in moisturization, wrinkles, pores, texture efficacy after 2 weeks, 4 weeks, 8 weeks of product use were assessed by instrumental testing.

4. The subject claims:

4.1 inclusion criteria

(1) Chinese female;

(2) age 22-55 years;

(3) dry or semi-dry skin;

(4) those with fine lines or wrinkles on the face;

(5) the subject should be insensitive to common cosmetics;

(6) subjects did not participate in additional clinical studies for the last 3 months;

4.2 exclusion criteria

(1) Antihistamines administered in the near week or immunosuppressants administered in the near month;

(2) any anti-inflammatory drug is used on the tested part in the last 2 months;

(3) insulin-dependent diabetic patients;

(4) patients suffering from respiratory diseases undergoing treatment;

(5) lactating or pregnant women;

(6) people with allergic constitution or allergic dermatitis have a history of skin diseases or diseases;

(7) a population undergoing dermatological treatment, or a subject taking within one month hydroxy-acids, whitening and anti-aging drugs;

(8) the skin characterization that the test region has large-area birthmarks, scratch marks, white spots, pigmented nevi, pimples and scars and the like which influence the experiment is carried out, and in addition, the responsible person of the experiment judges that the skin characterization is not suitable for the crowd as the experimental object.

4.3 limitations on items

(1) During the test, the treatment for the effect test on the skin to be tested is prohibited.

(2) During the test, retesting with other skin care products and other products that affect the test was prohibited.

(3) During the test period, daily products such as bath foam, soap, shampoo and the like are consistent with those used before the test is started, and replacement is prohibited.

4.4 target number of experiments: eligible subjects were enrolled for 35, ensuring a final completion of 30.

5. Testing an instrument:

(1) skin moisture tester (mileure intelligent skin moisture tester)

(2) Antera3D skin tester (Ireland)

(3) Canon E600 camera

(4) Balance: sensory quantity 0.01g (Mettler)

And (3) testing environment: the temperature is 25 +/-1 ℃, the humidity is 50% +/-5%, and the light rays are ensured to be unchanged every time.

6. The experimental process comprises the following steps:

(1) at the first visit, subjects were instructed to perform the trial and signed an informed consent.

(2) Screening the test subjects according to the test requirements, screening 35 subjects in the test group, and finally ensuring more than 30 subjects.

(3) Screening qualified subjects can not use any cosmetics in the morning, after the face is cleaned with clear water, the face is rested for 30 minutes in a constant temperature and humidity room, and after 30 minutes, the face is tested.

(4) And (3) carrying out use instructions on the testee after the test is finished, issuing the product after the testee listens to the instructions, using the test sample by the testee for 8 weeks continuously at home, carrying out return visit of the appointed place after 2 weeks, 4 weeks and 8 weeks of continuous use of the product according to the instruction of the test, and carrying out the face test according to the step (3). The amount of product used was confirmed at the beginning and end of the test.

7. And (3) observing test items:

subjects were tested 4 times 2 weeks, 4 weeks, 8 weeks after use prior to use:

(1) skin moisture was measured by using a mileure test 3 times and averaged, and the test position was near the temple on the right face of the volunteer.

And (4) judging a result: after the product is used, the moisture content in the test area is obviously increased, and the moisture retention effect of the test sample is shown.

(2) Skin wrinkles: wrinkle analysis with antera3D combined with canon camera image acquisition.

And (4) judging a result: the picture was taken with antera3D and wrinkles were analyzed.

(3) Skin texture: texture canon camera image acquisition was analyzed with antea 3D.

And (4) judging a result: the texture was analyzed by photographing with antera 3D.

8. The statistical method of the test results comprises the following steps:

TABLE 1 whether example Classification Table is adopted

TABLE 2 sample usage determination of whether to use case classification

Status of use	Whether to employ data
		During the test period, more than 3/4 are used	By using
During the test, the general method	By using
		Less than half of the test period	Conference for others
Is not substantially used	Without using
		In addition to the above-mentioned others,the administration and dosage do not comply with	Conference for others

9. The data statistical analysis method comprises the steps of carrying out descriptive statistics on each measured value by SPSS software, wherein the descriptive statistics comprises the number, the mean value, the standard deviation, the minimum value, the maximum value and the like, carrying out significance test on data improvement value normal distribution by Shapiro-wilk test, wherein Sig (double sides) > 0.01 shows normal distribution, carrying out T test, removing 0.05 from significance difference level α, and carrying out non-normal distribution if weak Sig (double sides) <0.01, carrying out Wilcoxon test, wherein the significance difference level α shows 0.05, the significance difference level is P value <0.05, which shows that the statistical difference exists, and P <0.01, and the statistical difference is obvious.

10. Analysis of Experimental results

10.1T group test item result data and analysis

(1) Summary of data for T-group test items 0 weeks (W0) and 8 weeks (W8)

TABLE 3 data of T-group test items at week 0 (W0) and week 8 (W8)

(2) Analysis of results of T-group experiments

The test results, 8 weeks after the use of placebo, showed that the group of subjects (30) had:

-moisture: compared with the prior art, after the test subjects of the group continuously use the product for 8 weeks, the product has certain moisturizing effect (P is 0.06), and the moisture increasing rate is 3.5%.

-wrinkles: compared with the prior use, after 8 weeks of continuous 8 weeks of the use of the subjects in the group, the total size of the wrinkles is reduced by 22 percent (P is less than or equal to 0.05), the depth of the wrinkles is reduced by 21 percent (P is 0.8),

-pore: there were no statistical differences at each time point.

-texture: there were no statistical differences at each time point.

10.2B group test item result data and analysis

(1) Summary of data in 0 week (W0) and 8 weeks (W8) of group B test items

TABLE 4 week 0 (W0) and week 8 (W8) group B test item data

P is less than or equal to 0.05, and P is less than or equal to 0.01. P.ltoreq.0.05 with statistical differences, the closer P is to 0, the higher the confidence.

(2) Analysis of group B test results

After the test population (33 persons) of this group, which used the wrinkle-removing tightening stock solution without recombinant collagen for B8 weeks, the test results showed that:

-moisture: compared with the prior art, after the test subjects of the group continuously use the product for 8 weeks, the product has certain moisturizing effect (P is less than or equal to 0.01), and the water lifting rate is 4.07%.

-wrinkles: compared with the prior use, the total size of the wrinkles is reduced by 32.43 percent (P is less than or equal to 0.01) and the depth of the wrinkles is reduced by 21.61 percent (P is less than or equal to 0.05) after the subjects in the group continuously use for 8 weeks.

-pore: the pore volume and the pore density have no statistical change.

-texture: there were no statistical differences.

10.3C group test item result data and analysis

(1) Summary of data for group C test items week 0 (W0) and week 8 (W8)

TABLE 5 week 0 (W0) and week 8 (W8) C test item data

(2) Analysis of group C Experimental results

The test results of this group of subjects (32) who used the wrinkle-removing tightening stock solution containing recombinant collagen for 8 weeks showed that:

moisture content: the water content was significantly changed at W8, the P value was less than 0.01, the statistical difference was observed, and the percentage improvement was 5.58%.

Wrinkling: the overall size of wrinkles and the depth of wrinkles were improved at week 8, with an overall wrinkle size improvement of 36.64% and a P value of less than 0.01, and an wrinkle depth improvement of 29.33% and a P value of 0.01 or less.

Pore: pore and pore density, no statistical difference.

Texture: the improvement is greater in week 8, the improvement rate is 8.7%, and the P value is less than 0.05, which has statistical significance.

In conclusion, the group B of the wrinkle-removing and tightening stock solution without recombinant collagen is superior to the group T of the placebo in moisture and wrinkles, which indicates that the wrinkle-removing and tightening stock solution has a certain wrinkle-removing effect. However, the group C is a test group containing the recombinant collagen wrinkle-removing and tightening stock solution, and the water and wrinkle-resisting effects of the group C are obviously superior to those of the group B and the group T of the control group which do not contain the recombinant collagen wrinkle-removing and tightening stock solution, so that the recombinant collagen has more obvious improvement effects on the overall size of wrinkles and the wrinkle depth, has obvious moisturizing effect and has better wrinkle-removing and tightening effects on the skin. The recombinant collagen fermentation source adopted in the embodiment ensures the safety of the product, has no peculiar smell of animal collagen, is close to the collagen lost in human skin, supplements the collagen in the skin, has good compatibility with skin tissues, and has good effects of removing wrinkles, tightening and moisturizing.

Claims (4)

1. The anti-wrinkle firming stock solution containing the recombinant collagen is characterized by comprising the following components in percentage by mass: 0.1-1% of recombinant collagen, 10-40% of butanediol, 1-5% of 1, 2-hexanediol, 81-10% of polyethylene glycol, 0.1-0.5% of sodium hyaluronate, 0.1-0.5% of polyglutamic acid, 0. 80.05-2% of acetyl hexapeptide and the balance of water.

2. The wrinkle-removing and tightening stock solution as claimed in claim 1, which comprises the following components by mass percent: 0.4% of recombinant collagen, 30% of butanediol, 2.5% of 1, 2-hexanediol, 0.4% of polyethylene glycol-810%, 0.4% of sodium hyaluronate, 0.4% of polyglutamic acid, 0-80.07% of acetyl hexapeptide and the balance of water.

3. The wrinkle-removing compact stock solution as set forth in claim 1, wherein the recombinant collagen is produced by fermentation of Pichia pastoris with accession No. CGMCC No. 5021.

4. The preparation method of the wrinkle-removing and tightening stock solution as claimed in claim 1, comprising the steps of:

adding sodium hyaluronate, polyglutamic acid and recombinant collagen into butanediol according to a certain proportion, stirring until uniformly mixing, then adding 1, 2-hexanediol, polyethylene glycol-8 and acetyl hexapeptide-8, adding water to 100%, stirring until completely dissolving, and obtaining the wrinkle-removing and tightening stock solution containing the recombinant collagen.

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