CN106176277A - Gene recombinant human lysozyme facial film and preparation method thereof - Google Patents
Gene recombinant human lysozyme facial film and preparation method thereof Download PDFInfo
- Publication number
- CN106176277A CN106176277A CN201610803396.2A CN201610803396A CN106176277A CN 106176277 A CN106176277 A CN 106176277A CN 201610803396 A CN201610803396 A CN 201610803396A CN 106176277 A CN106176277 A CN 106176277A
- Authority
- CN
- China
- Prior art keywords
- recombinant human
- lysozyme
- sodium
- phosphate buffer
- gene recombinant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/36—Carboxylic acids; Salts or anhydrides thereof
- A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/63—Steroids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/731—Cellulose; Quaternized cellulose derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/735—Mucopolysaccharides, e.g. hyaluronic acid; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/88—Two- or multipart kits
- A61K2800/882—Mixing prior to application
Abstract
The invention provides a kind of gene recombinant human lysozyme facial film, comprise: (1) lysozyme lyophilized powder, including gene recombinant human lysozyme and mannitol phosphate buffer, the ratio of the two is 0.5 0.8g: 1ml；(2) solvent, every 1000ml solvent includes hyaluronate sodium 0.35g 0.5g, low-numerator sodium hyaluronate 0.2g 0.6g, trimethyl glycine 3g 7g, sodium lactate 40ml 80ml, sorbitol 30ml 50ml, hydroxyethyl cellulose 1.5g 3g, glycyrrhizic acid dipotassium 1.5g 3g, EDTA 2Na 0.4g 0.7g and the sodium phosphate buffer of surplus.Present invention also offers the preparation method of gene recombinant human lysozyme facial film, including preparing the step of lysozyme lyophilized powder and preparing the step of solvent.
Description
Technical field
The invention belongs to cosmetic field, in particular it relates to a kind of facial film and preparation method thereof, particularly relate to
A kind of gene recombinant human lysozyme facial film and preparation method thereof.
Background technology
Facial film is a kind of special cosmetics, is coated on skin of face and forms thin film, by skin with extraneous every
Leaving, the effective ingredient in facial film can penetrate into skin, plays skin care, has additional nutrients, promotes that skin function and metabolism are made
With, so that skin of face is pure white, soft, pleasant, retain youthful vitality.At present, commercially available facial film goods are divided into by its form
Paste, liquid, glue and powdery.In terms of constituent, face mask of traditional Chinese medicine is in the majority, and activated protein facial film is the most rare.
Lysozyme (Lysozyme, LYZ), is a kind of to hydrolyze the alkaline enzyme of mucopolysaccharide in pathogenic bacterium, is widely present in people
In body Various Tissues, liquid and the microorganism such as birds and the Ovum Gallus domesticus album of poultry, the tear of mammal, saliva, blood plasma, urine, milk
In also containing this enzyme, wherein the abundantest with Ovum Gallus domesticus album content.Lysozyme is the most stable protein, has stronger heat resistance；Will not
Inactivating because of the process of organic solvent, when transferring in aqueous solution, vigor can full recovery；Can be frozen or be dried place
Reason, and vigor is stable；Antimicrobial spectrum is relatively wide, and safety is high.Anti-acne, reducing swelling and alleviating pain, degerming effect can be played for skin surface.
Summary of the invention
Present invention aims to the defect of prior art, it is provided that a kind of gene recombinant human lysozyme facial film and system thereof
Preparation Method.
On the one hand, the invention provides a kind of gene recombinant human lysozyme facial film, comprise:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and mannitol phosphate buffer, the ratio of the two is
0.5-0.8g gene recombinant human lysozyme: 1ml mannitol phosphate buffer；
(2) solvent, wherein, every 1000ml solvent includes hyaluronate sodium 0.35g-0.5g, low-numerator sodium hyaluronate
0.2g-0.6g, trimethyl glycine 3g-7g, sodium lactate 40ml-80ml, sorbitol 30ml-50ml, hydroxyethyl cellulose 1.5g-
3g, glycyrrhizic acid dipotassium 1.5g-3g, EDTA-2Na 0.4g-0.7g and the sodium phosphate buffer of surplus.
Aforesaid gene recombinant human lysozyme facial film, in lysozyme lyophilized powder, gene recombinant human lysozyme and mannitol
The proportioning of phosphate buffer is 0.68g gene recombinant human lysozyme: 1ml mannitol phosphate buffer.
Aforesaid gene recombinant human lysozyme facial film, every 1000ml solvent includes that hyaluronate sodium 0.45g, low molecule are transparent
Matter acid sodium 0.5g, trimethyl glycine 5g, sodium lactate 60ml, sorbitol 40ml, hydroxyethyl cellulose 2g, glycyrrhizic acid dipotassium 2g,
EDTA-2Na 0.5g and the sodium phosphate buffer of surplus.
Aforesaid gene recombinant human lysozyme facial film, described gene recombinant human lysozyme is made up of 130 aminoacid, its ammonia
Base acid sequence is: KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGI FQINSRYWCN
DGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV。
Aforesaid gene recombinant human lysozyme facial film, the pH value of described mannitol phosphate buffer be 6.5-7.2 (preferably
It is 7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L)；The matter of mannitol in described mannitol phosphate buffer
Amount percentage composition is 2%-5% (preferably 3%).
Aforesaid gene recombinant human lysozyme facial film, the molecular weight of described hyaluronate sodium is 120-150 ten thousand dalton；Institute
The molecular weight stating low-numerator sodium hyaluronate is 20-40 ten thousand dalton.
Aforesaid gene recombinant human lysozyme facial film, the pH value of described sodium phosphate buffer be 6.5-7.2 (preferably
7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L).
Aforesaid gene recombinant human lysozyme facial film, in use, first adds 1ml solvent in lysozyme lyophilized powder and dissolves
Sucking-off after uniformly, it is fed back to mix homogeneously in solvent, is then injected to soak on face paper.
On the other hand, the invention provides the preparation method of aforesaid gene recombinant human lysozyme facial film, including walking as follows
Rapid:
(1) lysozyme lyophilized powder is prepared
(1.1) 0.5g-0.8g gene recombinant human lysozyme is dissolved in aseptic mannitol phosphate buffer, subsequently
Filtration sterilization, obtains filtrate；
(1.2) adding aseptic mannitol phosphate buffer in filtrate and be settled to 100ml, mix homogeneously obtains bacteriolyze
Enzymatic solution；
(1.3) by lysozyme soln subpackage, lyophilizing, lysozyme lyophilized powder is obtained；
Wherein, above-mentioned steps (1.1) is aseptically carried out to (1.3)；
(2) solvent is prepared
(2.1) by transparent to 0.35g-0.5g (preferably 0.45g) hyaluronate sodium and 0.2g-0.6g (preferably 0.5g) low molecule
Matter acid sodium is dissolved in sodium phosphate buffer and obtains sodium hyaluronate solution；
(2.2) 1.5g-3g (preferably 2g) hydroxyethyl cellulose is dissolved in sodium phosphate buffer obtains hydroxyethyl cellulose
Solution；
(2.3) adding in hydroxyethyl cellulose solution by sodium hyaluronate solution, mix homogeneously obtains mixed solution；
(2.4) in mixed solution, 3g-7g (preferably 5g) trimethyl glycine, 1.5g-3g (preferably 2g) are added successively sweet
Oxalic acid dipotassium, 0.4g-0.7g (preferably 0.5g) EDTA-2Na, 40ml-80ml (preferably 60ml) sodium lactate and 30ml-50ml are (excellent
Select 40ml) sorbitol, it is subsequently added sodium phosphate buffer and is settled to 1000ml, mix homogeneously obtains solvent solutions；
(2.5) by solvent solutions subpackage, sterilizing, solvent is obtained.
Aforesaid preparation method, in step (1.1), the aqueous phase filter using aperture to be 0.22 μm carries out filtration sterilization.
Aforesaid preparation method, step (2.1) and (2.2) are carried out at 50-70 DEG C (preferably 60 DEG C).
Aforesaid preparation method, in step (2.5), described sterilizing is 121 DEG C of sterilizings 30 minutes.
Aforesaid preparation method, described gene recombinant human lysozyme is made up of 130 aminoacid, and its aminoacid sequence is:
KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNAC
HLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV。
Aforesaid preparation method, the pH value of described mannitol phosphate buffer is 6.5-7.2 (preferably 7.0), concentration
It is 4.5-6.0mmol/L (preferably 5mmol/L)；In described mannitol phosphate buffer, the weight/mass percentage composition of mannitol is
2%-5% (preferably 3%).
Aforesaid preparation method, the pH value of described sodium phosphate buffer is 6.5-7.2 (preferably 7.0), and concentration is 4.5
6.0mmol/L (preferably 5mmol/L).
Relative to prior art, gene recombinant human lysozyme facial film that the present invention provides and preparation method thereof has the most excellent
Point:
(1) the lysozyme raw material used is gene recombinant human lysozyme, uses biotechnology modified yeast pichia, logical
Crossing large scale fermentation, expression and purification to be prepared from, purity is high, activity is high, and safety is good, and cost is relatively low.
(2) overcome activated protein to preserve the deficiency being prone to degraded inactivation the most for a long time, be prepared as lyophilized powder, protect
The activity of card lysozyme, and without preservative.
Detailed description of the invention
For being fully understood by the purpose of the present invention, feature and effect, by following specific embodiment, the present invention is done in detail
Describe in detail bright.
In the majority with face mask of traditional Chinese medicine for facial film in the market, activated protein facial film is less, and undesirable the asking of effect
Topic, in first aspect, the invention provides a kind of gene recombinant human lysozyme facial film, and this gene recombinant human lysozyme facial film includes
Following component:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and mannitol phosphate buffer, the ratio of the two is
0.5-0.8g gene recombinant human lysozyme: 1ml mannitol phosphate buffer, it is preferable that the ratio of the two is 0.68g gene weight
Group human lysozyme: 1ml mannitol phosphate buffer.
Wherein, gene recombinant human lysozyme is to use gene engineering method to produce, and is homology with the lysozyme of human body,
It is made up of 130 aminoacid, and aminoacid sequence is:
KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNAC
HLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV.This lysozyme is
Host Strains, screening obtains expressing the pichia yeast genetic engineering bacteria of lysozyme, then produces by fermentation and obtain.The present invention uses
Gene recombinant human lysozyme be consistent with the lysozyme in human body, first, its biological homology, biocompatibility, biology
Safety is preferable；Secondly, gene recombinant human lysozyme has anti-acne, reducing swelling and alleviating pain, degermation, itself is exactly a kind of molten
Fungus matter, can play the effect of anticorrosion, sterilization, it is to avoid add the chemical preservative harm to human body.
Wherein, the pH value of mannitol phosphate buffer is 6.5-7.2 (preferably 7.0), and concentration is 4.5-6.0mmol/L
(preferably 5mmol/L)；In mannitol phosphate buffer, the weight/mass percentage composition of mannitol be 2%-5% (preferably
3%).In the freeze-drying process of gene recombinant human lysozyme, the mannitol phosphate of pH value used in the present invention and concentration delays
Rush liquid and can play the effect of protection lysozyme activity, simultaneously after lyophilizing, lysozyme in lyophilized powder can be made dispersed, homogeneous
Property high.
(2) solvent, including hyaluronate sodium, low-numerator sodium hyaluronate, trimethyl glycine, sodium lactate, sorbitol, hydroxyl
Ethyl cellulose, glycyrrhizic acid dipotassium, EDTA-2Na and sodium phosphate buffer, wherein, every 1000ml solvent includes hyaluronate sodium
0.35g-0.5g, low-numerator sodium hyaluronate 0.2g-0.6g, trimethyl glycine 3g-7g, sodium lactate 40ml-80ml, sorbitol
30ml-50ml, hydroxyethyl cellulose 1.5g-3g, glycyrrhizic acid dipotassium 1.5g-3g, EDTA-2Na 0.4g-0.7g and the phosphorus of surplus
Acid sodium buffer；Preferably, every 1000ml solvent includes hyaluronate sodium 0.45g, low-numerator sodium hyaluronate 0.5g, trimethyl
Glycine 5g, sodium lactate 60ml, sorbitol 40ml, hydroxyethyl cellulose 2g, glycyrrhizic acid dipotassium 2g, EDTA-2Na 0.5g and remaining
The sodium phosphate buffer of amount.
Wherein, the pH value of sodium phosphate buffer is 6.5-7.2 (preferably 7.0), concentration be 4.5 6.0mmol/L (preferably
It is 5mmol/L).The molecular weight of hyaluronate sodium is 120-150 ten thousand dalton, and the molecular weight of low-numerator sodium hyaluronate is 20-
400000 dalton.
After using above-mentioned specific components and specific proportioning, between each component of solvent, there is synergy, molten dissolving
It is more beneficial for dispersing and dissolving during bacterium enzyme lyophilized powder, after making facial film, the skin absorption to nutrition, and energy can be promoted in use
The activity keeping lysozyme of enough longer times.
Wherein, above-mentioned hyaluronate sodium, low-numerator sodium hyaluronate, trimethyl glycine, sodium lactate, sorbitol, hydroxyl
Ethyl cellulose, glycyrrhizic acid dipotassium, EDTA-2Na (i.e. disodiumedetate) all can conventional city available from.
Above-mentioned gene recombinant human lysozyme facial film in use, first takes a small amount of solvent (such as 1ml) and joins lysozyme
Lyophilized powder is uniformly dissolved rear sucking-off, and is fed back to mix homogeneously in solvent and obtains gene recombinant human lysozyme facial film solution, with
By this solution sucking-off and it is expelled on face paper with syringe afterwards, can use.For the ratio of lysozyme lyophilized powder Yu solvent,
Without particular/special requirement, as long as lysozyme lyophilized powder can fully be dissolved by solvent；In order to obtain higher cost performance with preferable
Using effect, the volume ratio of lysozyme lyophilized powder and solvent is preferably 1: 10-1: 15, and more preferably 1: 13.5.
The lysozyme component of above-mentioned gene recombinant human lysozyme facial film is prepared to lyophilized powder, and when stored with molten
Matchmaker is retained separately, and so ensure that the activity of lysozyme, it is to avoid activated protein preserves the most for a long time and to be prone to fall
The problem solving inactivation.
Second aspect, the invention provides the preparation method of said gene recombinant human lysozyme facial film, and the method includes system
The step of standby lysozyme lyophilized powder and the step preparing solvent, specific as follows:
The first step, prepares lysozyme lyophilized powder
First 0.5g-0.8g gene recombinant human lysozyme is dissolved in aseptic mannitol phosphate buffer (such as
Until being completely dissolved in 5ml), filtration sterilization subsequently, obtain filtrate.Wherein, filtration sterilization can use aqueous phase filter to carry out, example
As, the aqueous phase filter that aperture can be used to be 0.22 μm carries out filtration sterilization.
Adding aseptic mannitol phosphate buffer subsequently in above-mentioned filtrate and be settled to 100ml, mix homogeneously obtains
Lysozyme soln.
Subsequently by lysozyme soln subpackage, and use freeze dryer lyophilizing, thus obtain lysozyme lyophilized powder.According to actual need
, those skilled in the art can determine the volume of subpackage, such as, can be 1ml.
The above-mentioned step preparing lysozyme lyophilized powder is aseptically carried out, for example, it is possible at superclean bench
In aseptic complete.
The mannitol phosphate buffer used in this step can use conventional method to prepare, and its pH value is 6.5-7.2
(preferably 7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L)；In mannitol phosphate buffer, mannitol
Weight/mass percentage composition be 2%-5% (preferably 3%).According to the character of mannitol phosphate buffer, those skilled in the art
Suitable sterilizing methods can be selected, it is, for example possible to use aperture is the aqueous phase filter of the 0.45 μm mannitol phosphorus to preparing
Phthalate buffer filters, subsequently 121 DEG C of sterilizings 30 minutes.
Second step, prepares solvent
First by transparent to 0.35g-0.5g (preferably 0.45g) hyaluronate sodium and 0.2g-0.6g (preferably 0.5g) low molecule
Matter acid sodium is dissolved in sodium phosphate buffer (such as 200ml) and obtains sodium hyaluronate solution, in temperature 50-70 DEG C, and preferably 60 DEG C,
Mix homogeneously.
1.5g-3g hydroxyethyl cellulose is dissolved in sodium phosphate buffer (such as 300ml) subsequently and obtains hydroxy ethyl fiber
Cellulose solution, in temperature 50-70 DEG C, preferably 60 DEG C, mix homogeneously.
Adding in hydroxyethyl cellulose solution by sodium hyaluronate solution subsequently, mix homogeneously obtains mixed solution.Again to
Mixed solution is sequentially added into 3g-7g trimethyl glycine, 1.5g-3g glycyrrhizic acid dipotassium, 0.4g-0.7g EDTA-2Na, 40ml-
80ml sodium lactate and 30ml-50ml sorbitol, be subsequently added sodium phosphate buffer and be settled to 1000ml, and mix homogeneously obtains solvent
Solution；Preferably, it is sequentially added into 5g trimethyl glycine, 2g glycyrrhizic acid dipotassium, O.5g EDTA-2Na, 60ml to mixed solution
Sodium lactate and 40ml sorbitol, be subsequently added sodium phosphate buffer and be settled to 1000ml, and mix homogeneously obtains solvent solutions.
Subsequently by solvent solutions subpackage, and sterilizing, thus obtain solvent.According to actual needs, those skilled in the art's energy
Enough determine the volume of subpackage, such as, can be 13.5ml.Conventional method sterilizing can be used, such as 121 DEG C of sterilizings 30 minutes.
In this step use sodium phosphate buffer conventional method can be used to prepare, its pH value be 6.5-7.2 (preferably
7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L).Before use, first sodium phosphate buffer is filtered, example
As used the aqueous phase filter of 0.45 μm to filter.
Embodiment
Further illustrate the present invention below by the mode of embodiment, but the most therefore limit the present invention to described reality
Execute among example scope.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or according to business
Product description selects.Material used in embodiment is conventional commercial obtain.
Embodiment 1
The composition of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and pH value be 7.0, concentration be the mannitol of 5mmol/L
Phosphate buffer, the ratio of the two is 0.68g gene recombinant human lysozyme: 1ml mannitol phosphate buffer.
(2) solvent, wherein, every 1000ml solvent include hyaluronate sodium 0.45g, low-numerator sodium hyaluronate 0.5g, three
Methylglycine 5g, sodium lactate 60ml, sorbitol 40ml, hydroxyethyl cellulose 2g, glycyrrhizic acid dipotassium 2g, EDTA-2Na 0.5g
With the pH value of surplus be 7.0, concentration be the sodium phosphate buffer of 5mmol/L.
The preparation method of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder preparation: first join 5mmol/L pH7.0 phosphate buffer (the i.e. manna containing 3% mannitol
Alcohol phosphate buffer) 200ml, filters with the aqueous phase filter that aperture is 0.45 μm, then 121 DEG C, 30min sterilizing.Then claim
Amount gene recombinant human lysozyme dry powder 0.68g, the mannitol phosphate buffer adding 5ml sterilizing dissolves, and dissolves completely
Afterwards with the aqueous phase filter filtration sterilization of aperture 0.22 μm, then draw a small amount of mannitol phosphate buffer filtration and washing, then add
Mannitol phosphate buffer is settled to 100ml, is finally dispensed in aseptic cillin bottle, every bottle of 1ml, and whole operating process exists
Aseptic in super-clean bench complete, freeze dryer lyophilizing after subpackage.
(2) solvent preparation: the sodium phosphate buffer of the 5mmol/L pH7.0 of preparation 1200ml, is 0.45 μm with aperture
Aqueous phase filter filters.Then weigh hyaluronate sodium 0.45g, low-numerator sodium hyaluronate 0.5g, be dissolved in 200ml 5mmol/L
In the sodium phosphate buffer of pH7.0, stir in temperature 60 C standby.Weigh hydroxyethyl cellulose 2g, be dissolved in 300ml
In the sodium phosphate buffer of 5mmol/L pH7.0, after temperature 60 C stirs then molten by dissolving uniform hyaluronic acid
Liquid addition hydroxyethyl cellulose solution slowly continues to be stirred until homogeneous, adds trimethyl glycine 5g, glycyrrhizic acid dipotassium successively
2g, EDTA-2Na 0.5g, sodium lactate 60ml, sorbitol 40ml, with the sodium phosphate buffer of 5mmol/L pH7.0 after stirring
Liquid is settled to 1000ml, is again stirring for uniformly.Take the cillin bottle subpackage of 15ml, every bottle of 13.5ml, 121 DEG C, 30min sterilizing.
Before using, take lysozyme lyophilized powder one, draw with syringe and inhale after 1ml solvent addition lyophilized powder is uniformly dissolved
Go out, feed back in solvent and mix, the most again with the whole sucking-off of syringe, be expelled on face paper soak, it is thus achieved that gene recombinant human is molten
Bacterium enzyme facial film.
Embodiment 2
The composition of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and pH value be 6.5, concentration be the manna of 4.5mmol/L
Alcohol phosphate buffer, the ratio of the two is 0.5g gene recombinant human lysozyme: 1ml mannitol phosphate buffer.
(2) solvent, wherein, every 1000ml solvent include hyaluronate sodium 0.35g, low-numerator sodium hyaluronate 0.6g, three
Methylglycine 3g, sodium lactate 80ml, sorbitol 30ml, hydroxyethyl cellulose 1.5g, glycyrrhizic acid dipotassium 1.5g, EDTA-2Na
The sodium phosphate buffer that the pH value of 0.4g and surplus is 6.5, concentration is 4.5mmol/L.
The preparation method of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder preparation: first join the 4.5mmol/L pH6.5 phosphate buffer containing 2% mannitol (the sweetest
Dew alcohol phosphate buffer) 200ml, filters with the aqueous phase filter that aperture is 0.45 μm, then 121 DEG C, 30min sterilizing.Then
Weighing gene recombinant human lysozyme dry powder 0.5g, the mannitol phosphate buffer adding 5ml sterilizing dissolves, and dissolves completely
Afterwards with the aqueous phase filter filtration sterilization of aperture 0.22 μm, then draw a small amount of mannitol phosphate buffer filtration and washing, then add
Mannitol phosphate buffer is settled to 100ml, is finally dispensed in aseptic cillin bottle, every bottle of 1ml, and whole operating process exists
Aseptic in super-clean bench complete, freeze dryer lyophilizing after subpackage.
(2) solvent preparation: the sodium phosphate buffer of the 4.5mmol/L pH6.5 of preparation 1200ml, is 0.45 μm with aperture
Aqueous phase filter filter.Then weigh hyaluronate sodium 0.35g, low-numerator sodium hyaluronate 0.6g, be dissolved in 200ml
In the sodium phosphate buffer of 4.5mmol/L pH6.5, stir in temperature 60 C standby.Weigh hydroxyethyl cellulose 1.5g,
It is dissolved in the sodium phosphate buffer of 300ml 4.5mmol/L pH6.5, after temperature 60 C stirs, then will be uniformly dissolved
Hyaluronic acid solution slowly add in hydroxyethyl cellulose solution and continue to be stirred until homogeneous, add trimethyl glycine successively
3g, glycyrrhizic acid dipotassium 1.5g, EDTA-2Na 0.4g, sodium lactate 80ml, sorbitol 30ml, use 4.5mmol/ after stirring
The sodium phosphate buffer of LpH6.5 is settled to 1000ml, is again stirring for uniformly.Take the cillin bottle subpackage of 15ml, every bottle of 13.5ml,
121 DEG C, 30min sterilizing.
Before using, take lysozyme lyophilized powder one, draw with syringe and inhale after 1ml solvent addition lyophilized powder is uniformly dissolved
Go out, feed back in solvent and mix, the most again with the whole sucking-off of syringe, be expelled on face paper soak, it is thus achieved that gene recombinant human is molten
Bacterium enzyme facial film.
Embodiment 3
The composition of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and pH value be 7.2, concentration be the mannitol of 6mmol/L
Phosphate buffer, the ratio of the two is 0.5g gene recombinant human lysozyme: 1ml mannitol phosphate buffer.
(2) solvent, wherein, every 1000ml solvent includes hyaluronate sodium 0.5g, low-numerator sodium hyaluronate 0.2g, front three
Base glycine 7g, sodium lactate 40ml, sorbitol 50ml, hydroxyethyl cellulose 3g, glycyrrhizic acid dipotassium 3g, EDTA-2Na 0.7g and
The sodium phosphate buffer that the pH value of surplus is 7.2, concentration is 6mmol/L.
The preparation method of the gene recombinant human lysozyme facial film of the present embodiment is as follows:
(1) lysozyme lyophilized powder preparation: first join 6mmol/L pH7.2 phosphate buffer (the i.e. manna containing 5% mannitol
Alcohol phosphate buffer) 200ml, filters with the aqueous phase filter that aperture is 0.45 μm, then 121 DEG C, 30min sterilizing.Then claim
Amount gene recombinant human lysozyme dry powder 0.5g, the mannitol phosphate buffer adding 5ml sterilizing dissolves, after dissolving completely
With the aqueous phase filter filtration sterilization of aperture 0.22 μm, then draw a small amount of mannitol phosphate buffer filtration and washing, then add sweet
Dew alcohol phosphate buffer is settled to 100ml, is finally dispensed in aseptic cillin bottle, every bottle of 1ml, and whole operating process is super
Aseptic in clean platform complete, freeze dryer lyophilizing after subpackage.
(2) solvent preparation: the sodium phosphate buffer of the 6mmol/L pH7.2 of preparation 1200ml, is 0.45 μm with aperture
Aqueous phase filter filters.Then weigh hyaluronate sodium 0.5g, low-numerator sodium hyaluronate 0.2g, be dissolved in 200ml 6mmol/L
In the sodium phosphate buffer of pH7.2, stir in temperature 60 C standby.Weigh hydroxyethyl cellulose 3g, be dissolved in 300ml
In the sodium phosphate buffer of 6mmol/L pH7.2, after temperature 60 C stirs then molten by dissolving uniform hyaluronic acid
Liquid addition hydroxyethyl cellulose solution slowly continues to be stirred until homogeneous, adds trimethyl glycine 7g, glycyrrhizic acid dipotassium successively
3g, EDTA-2Na 0.7g, sodium lactate 40ml, sorbitol 50ml, with the sodium phosphate buffer of 6mmol/L pH7.2 after stirring
Liquid is settled to 1000ml, is again stirring for uniformly.Take the cillin bottle subpackage of 15ml, every bottle of 13.5ml, 121 DEG C, 30min sterilizing.
Before using, take lysozyme lyophilized powder one, draw with syringe and inhale after 1ml solvent addition lyophilized powder is uniformly dissolved
Go out, feed back in solvent and mix, the most again with the whole sucking-off of syringe, be expelled on face paper soak, it is thus achieved that gene recombinant human is molten
Bacterium enzyme facial film.
Claims (15)
1. a gene recombinant human lysozyme facial film, it is characterised in that comprise:
(1) lysozyme lyophilized powder, including gene recombinant human lysozyme and mannitol phosphate buffer, the ratio of the two is 0.5-
0.8g gene recombinant human lysozyme: 1ml mannitol phosphate buffer；
(2) solvent, wherein, every 1000ml solvent includes hyaluronate sodium 0.35g-0.5g, low-numerator sodium hyaluronate 0.2g-
0.6g, trimethyl glycine 3g-7g, sodium lactate 40ml-80ml, sorbitol 30ml-50ml, hydroxyethyl cellulose 1.5g-3g, sweet
Oxalic acid dipotassium 1.5g-3g, EDTA-2Na 0.4g-0.7g and the sodium phosphate buffer of surplus.
Gene recombinant human lysozyme facial film the most according to claim 1, it is characterised in that in lysozyme lyophilized powder, base
Because the proportioning of recombinant human lysozyme and mannitol phosphate buffer is 0.68g gene recombinant human lysozyme: 1ml mannitol phosphoric acid
Salt buffer.
Gene recombinant human lysozyme facial film the most according to claim 1, it is characterised in that every 1000ml solvent includes transparent
Matter acid sodium 0.45g, low-numerator sodium hyaluronate 0.5g, trimethyl glycine 5g, sodium lactate 60ml, sorbitol 40ml, ethoxy
Cellulose 2g, glycyrrhizic acid dipotassium 2g, EDTA-2Na 0.5g and the sodium phosphate buffer of surplus.
4. according to the gene recombinant human lysozyme facial film described in any one of claim 1-3, it is characterised in that described gene recombinaton
Human lysozyme is made up of 130 aminoacid, and its aminoacid sequence is:
KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRSTDYGIFQINSRYWCNDGKTPGAVNAC
HLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGCGV。
5. according to the gene recombinant human lysozyme facial film described in any one of claim 1-3, it is characterised in that described mannitol phosphorus
The pH value of phthalate buffer is 6.5-7.2 (preferably 7.0), and concentration is 4.5-6.0mmol/L (preferably 5mmol/L)；Described
In mannitol phosphate buffer, the weight/mass percentage composition of mannitol is 2%-5% (preferably 3%).
6. according to the gene recombinant human lysozyme facial film described in any one of claim 1-3, it is characterised in that described hyaluronic acid
The molecular weight of sodium is 120-150 ten thousand dalton；The molecular weight of described low-numerator sodium hyaluronate is 20-40 ten thousand dalton.
7. according to the gene recombinant human lysozyme facial film described in any one of claim 1-3, it is characterised in that described sodium phosphate delays
The pH value rushing liquid is 6.5-7.2 (preferably 7.0), and concentration is 4.5-6.0mmol/L (preferably 5mmol/L).
8. according to the gene recombinant human lysozyme facial film described in any one of claim 1-3, it is characterised in that in use, first
By 1ml solvent addition lysozyme lyophilized powder being uniformly dissolved rear sucking-off, being fed back to mix homogeneously in solvent, then it is injected to facial film
Soak on paper.
9. the preparation method of the gene recombinant human lysozyme facial film described in any one of claim 1-8, it is characterised in that include as
Lower step:
(1) lysozyme lyophilized powder is prepared
(1.1) 0.5g-0.8g gene recombinant human lysozyme is dissolved in aseptic mannitol phosphate buffer, filters subsequently
Degerming, obtain filtrate；
(1.2) adding aseptic mannitol phosphate buffer in filtrate and be settled to 100ml, it is molten that mix homogeneously obtains lysozyme
Liquid；
(1.3) by lysozyme soln subpackage, lyophilizing, lysozyme lyophilized powder is obtained；
Wherein, above-mentioned steps (1.1) is aseptically carried out to (1.3)；
(2) solvent is prepared
(2.1) by 0.35g-0.5g (preferably 0.45g) hyaluronate sodium and 0.2g-0.6g (preferably 0.5g) low molecular weight hyaluronic acid
Sodium is dissolved in sodium phosphate buffer and obtains sodium hyaluronate solution；
(2.2) 1.5g-3g (preferably 2g) hydroxyethyl cellulose is dissolved in sodium phosphate buffer obtains hydroxyethyl cellulose solution；
(2.3) adding in hydroxyethyl cellulose solution by sodium hyaluronate solution, mix homogeneously obtains mixed solution；
(2.4) in mixed solution, 3g-7g (preferably 5g) trimethyl glycine, 1.5g-3g (preferably 2g) glycyrrhizic acid are added successively
Dipotassium, 0.4g-0.7g (preferably 0.5g) EDTA-2Na, 40ml-80ml (preferably 60ml) sodium lactate and 30ml-50ml are (preferably
40ml) sorbitol, is subsequently added sodium phosphate buffer and is settled to 1000ml, and mix homogeneously obtains solvent solutions；
(2.5) by solvent solutions subpackage, sterilizing, solvent is obtained.
Preparation method the most according to claim 9, it is characterised in that in step (1.1), using aperture is 0.22 μm
Aqueous phase filter carry out filtration sterilization.
11. preparation methoies according to claim 9, it is characterised in that step (2.1) and (2.2) are at 50-70 DEG C (preferably
60 DEG C) carry out.
12. preparation methoies according to claim 9, it is characterised in that in step (2.5), described sterilizing is at 121 DEG C
Sterilizing 30 minutes.
13. according to the preparation method described in claim 9-12, it is characterised in that described gene recombinant human lysozyme is by 130
Aminoacid forms, and its aminoacid sequence is: KVFERCELARTLKRLGMDGYRGISLANWMCLAKWESGYNTRATNYNAGDRS
TDYGIFQINSRYWCNDGKTPGAVNACHLSCSALLQDNIADAVACAKRVVRDPQGIRAWVAWRNRCQNRDVRQYVQGC
GV。
14. according to the preparation method described in claim 9-12, it is characterised in that the pH value of described mannitol phosphate buffer
Being 6.5-7.2 (preferably 7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L)；Described mannitol phosphate-buffered
In liquid, the weight/mass percentage composition of mannitol is 2%-5% (preferably 3%).
15. according to the preparation method described in claim 9-12, it is characterised in that the pH value of described sodium phosphate buffer is 6.5-
7.2 (preferably 7.0), concentration is 4.5-6.0mmol/L (preferably 5mmol/L).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610803396.2A CN106176277A (en) | 2016-09-06 | 2016-09-06 | Gene recombinant human lysozyme facial film and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610803396.2A CN106176277A (en) | 2016-09-06 | 2016-09-06 | Gene recombinant human lysozyme facial film and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106176277A true CN106176277A (en) | 2016-12-07 |
Family
ID=58067341
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610803396.2A Pending CN106176277A (en) | 2016-09-06 | 2016-09-06 | Gene recombinant human lysozyme facial film and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106176277A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018120401A1 (en) * | 2016-12-29 | 2018-07-05 | 陕西慧康生物科技有限责任公司 | Cosmetic composition |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1410051A (en) * | 2002-06-05 | 2003-04-16 | 长春奇龙生物技术研究所 | Human lysozyme series cosmetics and its preparation method |
| CN1664096A (en) * | 2004-10-21 | 2005-09-07 | 北京双鹭立生医药科技有限公司 | Method for preparing human lysozyme |
| WO2008124764A1 (en) * | 2007-04-10 | 2008-10-16 | Saint Simeon Lda | Novel compositions containing lysozyme and c-1/c-4 polysaccharides and use thereof in oral care, cosmetology and dermatology, contraception, urology and gynecology |
| CN102188336A (en) * | 2011-05-16 | 2011-09-21 | 成都瑞商通银日用品有限公司 | Moisture retention and anti-aging compound |
| CN105194719A (en) * | 2015-10-28 | 2015-12-30 | 成都清科生物科技有限公司 | Facial wound restoration dressing and plaster, and preparation methods thereof |
-
2016
- 2016-09-06 CN CN201610803396.2A patent/CN106176277A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1410051A (en) * | 2002-06-05 | 2003-04-16 | 长春奇龙生物技术研究所 | Human lysozyme series cosmetics and its preparation method |
| CN1664096A (en) * | 2004-10-21 | 2005-09-07 | 北京双鹭立生医药科技有限公司 | Method for preparing human lysozyme |
| WO2008124764A1 (en) * | 2007-04-10 | 2008-10-16 | Saint Simeon Lda | Novel compositions containing lysozyme and c-1/c-4 polysaccharides and use thereof in oral care, cosmetology and dermatology, contraception, urology and gynecology |
| CN102188336A (en) * | 2011-05-16 | 2011-09-21 | 成都瑞商通银日用品有限公司 | Moisture retention and anti-aging compound |
| CN105194719A (en) * | 2015-10-28 | 2015-12-30 | 成都清科生物科技有限公司 | Facial wound restoration dressing and plaster, and preparation methods thereof |
Non-Patent Citations (2)
| Title |
|---|
| 孟繁智: "《新编精细化工应用配方10000例与生产工艺实用手册（上卷）》", 29 February 2004, 银声音像出版社 * |
| 陕西慧康生物科技有限责任公司: "基因重组人溶菌酶（LYZ）面膜", 《国产非特殊用途化妆品备案信息服务平台》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018120401A1 (en) * | 2016-12-29 | 2018-07-05 | 陕西慧康生物科技有限责任公司 | Cosmetic composition |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103243071B (en) | Clinical-grade human mesenchymal stem cell serum-free complete medium | |
| Chen et al. | In vitro cellular responses to scaffolds containing two microencapulated growth factors | |
| CN106754668B (en) | Stem cell culture solution and injection | |
| CN106318904B (en) | Application of mesenchymal stem cell conditioned medium in field of cosmetics | |
| CN106821938A (en) | A kind of preparation method of human mesenchymal stem cell freeze-dried powder | |
| CN108348555B (en) | Cell expansion methods and therapeutic compositions | |
| CN110564682B (en) | Method for large-scale production of human adipose-derived mesenchymal stem cell exosomes | |
| CN111297900B (en) | Composition rich in adipose-derived stem cell exosomes and application thereof | |
| Sordi et al. | Three-dimensional bioactive hydrogel-based scaffolds for bone regeneration in implant dentistry | |
| CN106913583A (en) | The preparation method and application of human mesenchymal stem cell source excretion body biologically active agents | |
| CN105079859A (en) | Dressing and preparation method thereof | |
| CN102597230A (en) | Particle-containing cell aggregate | |
| JP2020505442A (en) | Injectable composition for preventing hair loss or promoting hair growth | |
| CN104434791A (en) | Preparation and application of modified bletilla striata polysaccharide derivative nano-carrier | |
| CN110240627A (en) | A method of small active peptides being extracted from pearl powder using complex enzyme | |
| CN101020715B (en) | Process of extracting and preparing deer nerve growth factor (DEER NGF) | |
| CN101544683A (en) | Method and substance for keeping fibrinogen activity in thermal treatment | |
| CN106619175A (en) | Genetic recombination superoxide dismutase mask | |
| Zocchi et al. | New perspectives in regenerative medicine and surgery: the bioactive composite therapies (BACTs) | |
| WO2006134951A1 (en) | Long-term culture proliferation method for fat-derived stem cell | |
| CN106176277A (en) | Gene recombinant human lysozyme facial film and preparation method thereof | |
| Gou et al. | Adipose-derived mesenchymal stem cells (MSCs) are a superior cell source for bone tissue engineering | |
| CN107236032B (en) | Method for extracting compound cell factor from umbilical cord tissue | |
| CN113957040A (en) | Adipose-derived stem cell growth factor extract and preparation method and application thereof | |
| CN104707173A (en) | Three-dimensional cell culture platform, as well as preparation method and application method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161207 |
|
| RJ01 | Rejection of invention patent application after publication |