Disclosure of Invention
In view of the shortcomings in the prior art, the inventor of the present invention has long studied and practiced in a large number of ways to propose the technical scheme of the present invention. The technical scheme, the implementation process, the principle and the like are further explained as follows.
The technical scheme of the invention is as follows: the invention provides a recombinant human collagen mask with repairing activity and a preparation method thereof, wherein the recombinant human collagen mask comprises a close-fitting mask carrier and a mask material body, and raw materials for preparing the mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methyl parahydroxybenzoate, sodium propyl parahydroxybenzoate, phenoxyethanol, glycerol and purified water.
In some embodiments of the present invention, the mask material comprises the following components in parts by weight: 3-10 parts of recombinant humanized collagen solution; 1-5 parts of an efficacy promoting composition; 0.01-0.5 part of hydroxyethyl cellulose; 0.01-0.5 parts of carbomer; 0.01-0.2 part of sodium methylparaben; 0.01-0.2 part of sodium propyl parahydroxybenzoate; 0.1-0.8 part of phenoxyethanol; 1-5 parts of glycerol; purifying the water balance.
Preferably, the mask material comprises the following components in parts by weight: 5-10 parts of recombinant humanized collagen solution; 1-3 parts of an efficacy promoting composition; 0.05-0.25 part of hydroxyethyl cellulose; 0.05-0.15 parts of carbomer; 0.03-0.13 part of sodium methylparaben; 0.02-0.1 part of sodium propyl parahydroxybenzoate; 0.2-0.5 part of phenoxyethanol; 3-5 parts of glycerol; purifying the water balance.
Preferably, the recombinant humanized collagen solution is contained at a concentration of 1-15mg/mL.
Preferably, the close-fitting mask carrier is selected from one of non-woven fabrics, silk or biological fibers.
In some embodiments of the invention, the actuation composition comprises the following components in parts by weight: 1-5 parts of gamma-oryzanol; 0.1-0.8 part of bird's nest acid; 0.1-2 parts of butterflybush flower extract.
Preferably, the actuation composition comprises the following components in parts by weight: 1-3 parts of gamma-oryzanol; 0.1-0.5 part of bird's nest acid; 0.5-2 parts of butterflybush flower extract.
Preferably, the preparation method of the buddleja officinalis extract comprises the following steps: soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under atmospheric pressure for 3-4h, saidThe weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
Further, the preparation method of the recombinant humanized collagen solution comprises the following steps: preparing 10-12g/L of recombinant human collagen into a solution, adjusting the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain the recombinant humanized collagen solution.
Preferably, the molecular weight of the recombinant humanized collagen solution is in the range of 204-2530Da.
In some embodiments of the present invention, the present invention further provides a method for preparing a recombinant humanized collagen mask having a repair activity, the method comprising the steps of:
(1) Weighing hydroxyethyl cellulose, carbomer, sodium methyl parahydroxybenzoate, sodium propyl parahydroxybenzoate, phenoxyethanol, glycerol and purified water, stirring at 80deg.C for dissolving, sterilizing at 115deg.C for 30min, and cooling to below 40deg.C to obtain mixture A;
(2) Filtering the recombinant humanized collagen solution through a 0.22 mu m filter membrane, adding the filtered recombinant humanized collagen solution into the mixture A in the step (1), and stirring and mixing to obtain a mixture B;
(3) Adding the actuation composition into the mixture B, stirring and mixing to obtain a mask material, and filling the mask material into a sterile film bag;
(4) Folding and putting the cling type facial mask carrier into 25g of facial mask material body for soaking, and sealing to obtain the recombinant human collagen facial mask.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a recombinant human collagen facial mask with repairing activity and a preparation method thereof, wherein after the recombinant human collagen facial mask is used, the moisture content of skin cuticle is improved, the redness area a value, the transepidermal moisture loss value and the skin heme value of skin are reduced, so that the recombinant human collagen facial mask can be used for repairing skin barriers, improving the moisture content of skin, and relieving and improving the redness state of skin;
the recombinant human collagen III sequence of the active ingredient recombinant humanized collagen solution in each mask is a sequence optimized by long-term screening, the preparation method is described in Chinese patent application No. 107033238B, which is a purification method and a preparation method of recombinant human collagen III, and the fermentation liquor obtained by fermenting recombinant saccharomycetes capable of expressing the recombinant human collagen III is subjected to heat treatment at the temperature of 65-75 ℃, so that the degradation of the recombinant human collagen III in the fermentation liquor into small molecular peptides is effectively prevented, the production of endotoxin in the fermentation liquor is reduced, and the purity of the recombinant human collagen III is greatly improved;
the recombinant human III type collagen has strong water solubility and biological activity, has hydrophilic amino acid residues, has strong hydrophilicity and adhesiveness, and is favorable for the adhesion growth of fibroblasts; in a short time, a microenvironment is created for the cells, the cells are guided to quickly enter the damaged part of the skin to be repaired for growth, and then the skin barrier is repaired; the buddleja officinalis extract in the promotion composition is rich in sodium hyaluronate and glycoside substances, and can promote the efficacy of a recombinant humanized collagen solution by combining with gamma-oryzanol and nidus Collocaliae acid, and the promotion composition can cooperatively repair skin barrier, effectively lock water and promote the synthesis of human skin collagen;
in addition, the hydroxyethyl cellulose has good solubility and viscosity, can keep the original shape of the cosmetics in seasons with alternating cold and hot, and has the characteristic of moisture preservation; carbomers, which are commonly used thickeners in cosmetics, have a good affinity for the skin, increasing the viscosity of the cosmetics; sodium methylparaben and sodium propylparaben are common preservatives for cosmetics, have broad-spectrum balanced inhibition effects on bacteria, saccharomycetes and moulds, are typical antibacterial and antiseptic products, and have certain bactericidal capacity; the phenoxyethanol acts on the cell membrane to increase the passing rate of potassium ions and reduce the activity of enzyme, so that the cell growth is inhibited, the antibacterial effect is achieved, and meanwhile, the preservative effect is achieved;
meanwhile, the carrier of the clinging type facial mask adopts the lightest and fine silk, and can cling to skin textures, so that the clinging degree of the facial mask on the face is very high, the nose wing and mouth corners can be clinged, the nose wing and mouth corners are transparent and breathable, water locking and moisturizing are durable and deep, the silk facial mask can accelerate the repair of the skin, and the damaged skin can be quickly repaired through fine clinging of silk fibers.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the invention; all other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the present invention. The preferred methods and materials described herein are illustrative only and should not be construed as limiting the scope of the present application.
The experimental methods in the following examples are all conventional methods unless otherwise specified; the test materials and test strains used in the examples described below, unless otherwise specified, were commercially available.
Example 1
Recombinant human collagen mask with repairing activity and preparation method thereof
The recombinant human collagen facial mask comprises a cling type facial mask carrier and a facial mask material body, wherein raw materials for preparing the facial mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methylparaben, sodium propylparaben, phenoxyethanol, glycerol and purified water.
The mask material body comprises the following components in parts by weight (g/part): 10 parts of recombinant humanized collagen solution; 1 part of an efficacy promoting composition; 0.1 parts of hydroxyethyl cellulose; 0.15 parts of carbomer; 0.04 parts of sodium methylparaben; 0.02 parts of sodium propyl parahydroxybenzoate; 0.3 parts of phenoxyethanol; 3 parts of glycerol; 85.39 parts of purified water.
The concentration of the recombinant humanized collagen solution is 8mg/mL.
The cling type facial mask carrier is selected from one of non-woven fabrics.
The promoting composition comprises the following components in parts by weight: 1 part of gamma-oryzanol; 0.1 part of bird's nest acid; 0.5 part of butterflybush flower extract.
The preparation method of the buddleja officinalis extract comprises the following steps: soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under air pressure for 3h, wherein the weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
The preparation method of the recombinant humanized collagen solution comprises the following steps: preparing a solution of 10g/L recombinant human collagen, regulating the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain a recombinant humanized collagen solution, wherein the molecular weight of the recombinant humanized collagen solution is 204-2530Da.
The invention also provides a preparation method of the recombinant human collagen facial mask with repairing activity, which comprises the following steps: (1) Weighing hydroxyethyl cellulose, carbomer, sodium methyl parahydroxybenzoate, sodium propyl parahydroxybenzoate, phenoxyethanol, glycerol and purified water, stirring at 80deg.C for dissolving, sterilizing at 115deg.C for 30min, and cooling to below 40deg.C to obtain mixture A; (2) Filtering the recombinant humanized collagen solution through a 0.22 mu m filter membrane, adding the filtered recombinant humanized collagen solution into the mixture A in the step (1), and stirring and mixing to obtain a mixture B; (3) Adding the synergistic composition into the mixture B, stirring and mixing to obtain a mask material body, and filling the mask material body into a sterile film bag; (4) Folding and putting the cling type facial mask carrier into 25g of facial mask material body for soaking, and sealing to obtain the recombinant human collagen facial mask.
Example 2
Recombinant human collagen mask with repairing activity and preparation method thereof
The recombinant human collagen facial mask comprises a cling type facial mask carrier and a facial mask material body, wherein raw materials for preparing the facial mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methylparaben, sodium propylparaben, phenoxyethanol, glycerol and purified water.
The mask material body comprises the following components in parts by weight (g/part): 5 parts of recombinant humanized collagen solution; 3 parts of an efficacy promoting composition; 0.2 parts of hydroxyethyl cellulose; 0.1 parts of carbomer; 0.1 part of sodium methylparaben; 0.02 parts of sodium propyl parahydroxybenzoate; 0.2 parts of phenoxyethanol; 5 parts of glycerol; 86.38 parts of purified water.
The concentration of the recombinant humanized collagen solution is 15mg/mL.
The cling type mask carrier is selected from one of silk.
The promoting composition comprises the following components in parts by weight: 3 parts of gamma-oryzanol; 0.5 parts of bird's nest acid; 2 parts of butterflybush flower extract.
Soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under air pressure for 4h, wherein the weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
The preparation method of the recombinant humanized collagen solution comprises the following steps: preparing a solution of 12g/L recombinant human collagen, regulating the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain a recombinant humanized collagen solution, wherein the molecular weight of the recombinant humanized collagen solution is 204-2530Da.
The invention also provides a preparation method of the recombinant human collagen facial mask with repairing activity, which is carried out according to the embodiment 1.
Example 3
Recombinant human collagen mask with repairing activity and preparation method thereof
The recombinant human collagen facial mask comprises a cling type facial mask carrier and a facial mask material body, wherein raw materials for preparing the facial mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methylparaben, sodium propylparaben, phenoxyethanol, glycerol and purified water.
The mask material body comprises the following components in parts by weight (g/part): 8 parts of recombinant humanized collagen solution; 2 parts of an efficacy promoting composition; 0.2 parts of hydroxyethyl cellulose; 0.05 parts of carbomer; 0.1 part of sodium methylparaben; 0.1 part of sodium propyl parahydroxybenzoate; 0.3 parts of phenoxyethanol; 5 parts of glycerol; 84.25 parts of purified water.
The concentration of the recombinant humanized collagen solution is 1mg/mL.
The cling type facial mask carrier is selected from biological fibers.
The promoting composition comprises the following components in parts by weight: 2 parts of gamma-oryzanol; 0.3 parts of bird's nest acid; 1.2 parts of butterflybush flower extract.
Soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under air pressure for 3h, wherein the weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
The preparation method of the recombinant humanized collagen solution comprises the following steps: preparing a solution of 10g/L recombinant human collagen, regulating the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain a recombinant humanized collagen solution, wherein the molecular weight of the recombinant humanized collagen solution is 204-2530Da.
The invention also provides a preparation method of the recombinant human collagen facial mask with repairing activity, which is carried out according to the embodiment 1.
Example 4
Recombinant human collagen mask with repairing activity and preparation method thereof
The recombinant human collagen facial mask comprises a cling type facial mask carrier and a facial mask material body, wherein raw materials for preparing the facial mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methylparaben, sodium propylparaben, phenoxyethanol, glycerol and purified water.
The mask material body comprises the following components in parts by weight (g/part): 10 parts of recombinant humanized collagen solution; 1 part of an efficacy promoting composition; 0.25 parts of hydroxyethyl cellulose; 0.1 parts of carbomer; 0.13 parts of sodium methylparaben; 0.02 parts of sodium propyl parahydroxybenzoate; 0.2 parts of phenoxyethanol; 4 parts of glycerol; 84.3 parts of purified water.
The concentration of the recombinant humanized collagen solution is 5mg/mL.
The cling type facial mask carrier is selected from silk.
The promoting composition comprises the following components in parts by weight: 3 parts of gamma-oryzanol; 0.3 parts of bird's nest acid; 1 part of buddleja officinalis extract.
Soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under air pressure for 3h, wherein the weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
The preparation method of the recombinant humanized collagen solution comprises the following steps: preparing a solution of 10g/L recombinant human collagen, regulating the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain a recombinant humanized collagen solution, wherein the molecular weight of the recombinant humanized collagen solution is 204-2530Da.
The invention also provides a preparation method of the recombinant human collagen facial mask with repairing activity, which is carried out according to the embodiment 1.
Example 5
Recombinant human collagen mask with repairing activity and preparation method thereof
The recombinant human collagen facial mask comprises a cling type facial mask carrier and a facial mask material body, wherein raw materials for preparing the facial mask material body comprise a recombinant humanized collagen solution, hydroxyethyl cellulose, carbomer, an effect-promoting composition, sodium methylparaben, sodium propylparaben, phenoxyethanol, glycerol and purified water.
The mask material body comprises the following components in parts by weight (g/part): 5 parts of recombinant humanized collagen solution; 2 parts of an efficacy promoting composition; 0.05 parts of hydroxyethyl cellulose; 0.15 parts of carbomer; 0.03 parts of sodium methylparaben; 0.1 part of sodium propyl parahydroxybenzoate; 0.5 part of phenoxyethanol; 3 parts of glycerol; 89.17 parts of purified water.
The concentration of the recombinant humanized collagen solution is 10mg/mL.
The cling type facial mask carrier is selected from silk.
The promoting composition comprises the following components in parts by weight: 2 parts of gamma-oryzanol; 0.3 parts of bird's nest acid; 1 part of buddleja officinalis extract.
Soaking flos Buddlejae powder in water at a weight ratio of 1:10, extracting at 40deg.C under stirring for 1 hr, and adding lactic acid in CO2 Extracting under air pressure for 3h, wherein the weight ratio of the powder of the buddleja officinalis to the lactic acid is 1:5, filtering the extract, and concentrating under reduced pressure to obtain the buddleja officinalis extract.
The preparation method of the recombinant humanized collagen solution comprises the following steps: preparing a solution of 10g/L recombinant human collagen, regulating the pH to 8-9, and adding protease with the volume ratio of 0.1% and trypsin with the volume ratio of 0.1% for hydrolysis for 3 hours; and inactivating enzyme after the reaction is finished, centrifuging, collecting supernatant, and freeze-drying to obtain a recombinant humanized collagen solution, wherein the molecular weight of the recombinant humanized collagen solution is 204-2530Da.
The invention also provides a preparation method of the recombinant human collagen facial mask with repairing activity, which is carried out according to the embodiment 1.
Comparative example 1
This comparative example provides a recombinant human collagen mask having a repair activity and a method for preparing the same, which is different from example 4 only in that no agonist composition is contained in all the components, and the remaining components and the content of the components are the same as example 4.
Comparative example 2
This comparative example provides a recombinant humanized collagen mask having a repair activity and a method for preparing the same, which is different from example 4 only in that the recombinant humanized collagen solution is not contained in all components, and the remaining components and the content of the components are the same as example 4.
Performance testing
In order to verify the repairing performance of the recombinant human collagen facial mask prepared by the invention, the recombinant human collagen facial masks prepared by the example 4, the comparative example 1 and the comparative example 2 are taken and sent to Hangzhou Sieve quality technical service Co., ltd for testing.
The testing method and the testing process are as follows: collecting qualified subjects, cleaning the face of the subjects by using a facial cleaning product of a laboratory under the guidance of a detection personnel, sucking the face by using a chipless water-absorbing dry paper towel after the face is washed clean, sitting for 20 minutes in a rest waiting area with the temperature of 21+/-1 ℃ and the relative humidity of 50+/-10%, exposing and keeping the face relaxed, and measuring the numerical value before use by using a method shown in a table 1; the subject uses the recombinant human collagen mask prepared by the invention to measure the transepidermal water loss value, the skin horny layer water content, the skin heme value and the redness area a of the face skin of the subject after 14 and 28 days later* Values, data before use, 14 days use, 28 days use, test environment: temperature: 20-22 ℃, relative humidity: 40% -60% of the test indexes and the instruments are shown in table 1.
Inclusion criteria: 1) Healthy interviewees (except pregnant or lactating women) between 18 and 60 years of age; 2) Value of transepidermal water loss at cheek and cheekbone>15g/h/m2 And (3) the person; 3) The facial skin is easy to dry, itchy, reddish, fragile, sensitive and rough; 4) The face skin has no obvious skin damage, scar and other conditions; 5) The test can be well matched according to the requirements of the scheme, and the regularity of life can be maintained during the test; 6) Voluntary participation in the trial is able to read and understand all of the content of the informed consent form and sign the informed consent form.
Exclusion criteria: 1) Antihistamines used in the last week or immunosuppressants used in the last month; 2) Patients with skin history such as psoriasis, eczema, atopic dermatitis, severe acne, etc., or other determinants that may affect the experimental results; 3) Allergic diseases and allergic history of cosmetics or other external preparations; 4) Facial cosmetic surgery, other cosmetic forms or topical medications were received in nearly three months; 5) Insulin dependent diabetes mellitus patients; 6) Asthma or other chronic respiratory disease patients undergoing treatment; 7) Patients who received anti-cancer chemotherapy for nearly six months; 8) Patients with immunodeficiency or autoimmune disease; 9) Women in lactation or gestation, or those who have a pregnancy preparation plan recently; 10 A) are participating in or nearly two months in other clinical trials; 11 A person with high sensitivity to physical constitution; 12 A person who cannot complete the prescribed content according to the test requirement; 13 In addition to the above, it is judged by the staff that it is not suitable for the efficacy tester.
Table 1 skin repair test index
The data statistical analysis method comprises the following steps: carrying out descriptive statistics on the measured values of all the test areas by using SPSS software; significance Test of normal distribution of data improvement values was performed with Shapiro-Wilk Test, which indicated that the level of significance difference reached 0.05, with significant difference; * Indicating significant difference levels up to 0.01, the difference was very significant.
As shown in fig. 1, the moisture content of the skin cuticle of the recombinant humanized collagen mask of example 4 is significantly improved, the skin barrier function is better, and the difference change of comparative example 2 is not obvious, which indicates that the recombinant humanized collagen solution can adjust the skin moisture content, and compared with comparative example 1, the effect of example 4 is better, which indicates that the effect promoting composition has the effect of promoting the recombinant humanized collagen solution to play a role, and as shown in fig. 2, 3 and 4, the redness area a, the transepidermal moisture loss value and the skin heme value of the skin can be reduced after the recombinant humanized collagen mask of example 4 is used, so that the recombinant humanized collagen mask prepared by the invention can repair and care the skin barrier, improve the skin moisture content, and relieve and improve the redness state of the skin.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
The invention and its embodiments have been described above with no limitation, and the invention is illustrated in the figures of the accompanying drawings as one of its embodiments, without limitation in practice. In summary, those skilled in the art, having benefit of this disclosure, will appreciate that the invention can be practiced without the specific details disclosed herein.