CN114796594A - Preparation method and application of yeast recombinant collagen liquid dressing - Google Patents
Preparation method and application of yeast recombinant collagen liquid dressing Download PDFInfo
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- CN114796594A CN114796594A CN202210350425.XA CN202210350425A CN114796594A CN 114796594 A CN114796594 A CN 114796594A CN 202210350425 A CN202210350425 A CN 202210350425A CN 114796594 A CN114796594 A CN 114796594A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0028—Polypeptides; Proteins; Degradation products thereof
- A61L26/0033—Collagen
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/41—Anti-inflammatory agents, e.g. NSAIDs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/45—Mixtures of two or more drugs, e.g. synergistic mixtures
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a preparation method and application of yeast recombinant collagen liquid dressing, which is characterized by comprising the following raw and auxiliary materials in parts by weight: 8-15 parts of yeast recombinant collagen, 6-10 parts of a traditional Chinese medicine active monomer composition, 1-9 parts of sodium hyaluronate, 8-15 parts of glycerol, 10-20 parts of propylene glycol, 1-2 parts of phosphate buffer solution and the balance of purified water. Compared with the prior art, the traditional Chinese medicine active monomer composition and the yeast recombinant collagen are used together, the curative effect is definite, the traditional Chinese medicine active monomer composition is safe and non-irritant, has the functions of resisting bacteria and diminishing inflammation, promoting wound healing, reducing transepidermal water loss and the like, and has obvious curative effects on skin barrier function damage, dermatitis, eczema, facial acne and other diseases.
Description
Technical Field
The invention relates to the technical field of medicines, and particularly relates to a preparation method and application of a yeast recombinant collagen liquid dressing.
Background
Skin barrier function damage, dermatitis, eczema and facial acne are clinically common skin diseases, and the skin diseases are closely related to factors such as hyperseborrhea, follicular sebaceous gland duct blockage, bacterial infection and the like. Acne (including closed and open comedo), pimple, pustule, nodule and cyst are frequently accompanied by pruritus, red swelling and increased seborrhea, and the serious acne is suppurative, is better occurred in young men and women, and is inappropriately treated to easily generate permanent injury. For the skin diseases, the western medicine needle generally adopts tretinoin cream or gel clinically, benzoyl peroxide gel is also commonly used, and the oral antibiotics are matched for moderate acne. Severe oral administration of isotretinoin is required, although western medicine can relieve symptoms, it may cause permanent damage to the skin and endocrine dyscrasia. The traditional Chinese medicine for treating the skin diseases has the advantages of good curative effect and small side effect, can be used for local treatment, and has no side effect on the whole body. The purposes of preventing relapse and formation of pox scars, pockmarks and scars are achieved.
Compared with the traditional Chinese medicine decoction pieces, the Chinese medicine monomer with definite activity can avoid contact of insoluble impurities and the skin surface, so that the content of soluble impurities is greatly reduced, the curative effect is more exact and obvious, and the side effect is less. At present, a plurality of traditional Chinese medicine monomers have definite effects, such as ligustilide, ferulic acid, pachymin, angelone and the like, all have bacteriostatic action in vitro, and have obvious treatment effects on skin diseases such as acne, dermatitis and the like. In addition, the Chinese medicinal monomers also have the effects of resisting oxidation and melanin precipitation, and have a repairing effect on pigmentation caused by acne, dermatitis and the like.
Collagen is a protein family which is most abundant in animals and is an important component for supporting and protecting connective tissues of organisms, but the content of natural collagen is low, the extraction is difficult, the yeast recombinant collagen has the advantages of high yield, short production period, simple culture, easy acquisition of high-density fermentation and the like, has the advantages of no pyrogen, extracellular secretion of products, simple purification, post-translational processing and modification, high genetic stability of exogenous genes and the like, has activity and conformation similar to those of natural protein, and can make vessel wall elastic, prevent embolism, improve mechanical properties of cartilage and ligament, etc., the effects of the medicine on the aspects of skin injury repair, plastic and beauty, body immunity enhancement and the like are also commonly known, meanwhile, the cosmetic contains a large amount of polar groups, glycine and other natural moisturizing factors, and has the effect of promoting tissue metabolism.
At present, in the process of treating skin diseases, external preparations are often used, but the traditional external preparations are difficult to inhibit the propagation of pathogenic bacteria, easily cause the expansion of affected parts and further cause new diseases. According to the invention, the traditional Chinese medicine active monomer composition and the yeast recombinant collagen are matched for use, and the liquid dressing is prepared after auxiliary materials such as phosphate buffer solution and the like are added, so that the liquid dressing can play a role of protection when being laid on the skin, and can be used as a barrier to avoid external environmental pollution. It also has effects in inhibiting pathogen diffusion, promoting skin repair, accelerating wound healing, and effectively preventing scar formation and pigmentation.
Disclosure of Invention
The invention aims to provide a yeast recombinant collagen liquid dressing.
In order to achieve the above purpose, the invention provides a yeast recombinant collagen liquid dressing for treating skin diseases, which comprises the following specific embodiments:
(1) taking 10-20 parts of propylene glycol, heating to 50 ℃, slowly adding the traditional Chinese medicine active monomers in sequence according to the proportion, stirring until the traditional Chinese medicine active monomers are completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 1-9 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Adding 8-15 parts of yeast recombinant collagen into the sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 8-15 parts of glycerol, continuously stirring for 45min, and standing for 1h at constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 1-3 parts of phosphate buffer solution to adjust the pH value, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the traditional Chinese medicine active monomer composition in the step (1) is as follows: 1-3 parts of ligustilide, 1-6 parts of ferulic acid, 4-7 parts of angelone and 3-8 parts of poria cocos.
And (4) adding a phosphate buffer solution to adjust the pH value to 6-8.
Ligustilide is the main active ingredient of Chinese traditional medicine, Umbelliferae plant angelica volatile oil. Has strong pharmacological action on circulatory system and immunologic function, and can improve microcirculation, relax smooth muscle, inhibit bacteria, and improve immunoregulation function of organism. Has effects of inhibiting various pathogenic microorganisms causing skin diseases, whitening skin, and removing speckle.
Ferulic acid is one of the effective components in ferula asafetida. Has cis-isomer and trans-isomer, and is light yellow solid. The ferulic acid has a remarkable inhibiting effect on inflammatory proteins, and can reduce the activity of xanthine oxidase, further kill pathogenic bacteria and inhibit the inflammatory proteins. And ferulic acid can eliminate free radicals, has remarkable oxidation resistance, has a good scavenging effect on superoxide radicals and the like, can delay skin aging and prevent the formation of color spots.
Angelone, which is also an effective component of angelica, has an in vitro antibacterial effect, has a strong inhibitory effect against pathogenic bacteria of various skin diseases, can promote the generation of hemoglobin and erythrocytes, can dilate blood vessels, improve peripheral circulation, and promote skin repair, and is widely used in external preparations.
The poria cocos extract is a main effective component in the traditional Chinese medicine poria cocos, and researches show that the poria cocos extract can enhance the lymphocyte toxicity by 20-28 times in an in vitro test and obviously increase the content of immunoglobulin igG. Has inhibitory effect on contact skin allergic reaction, and has remarkable therapeutic effect on various skin diseases.
Compared with the prior art, the invention has the following beneficial effects:
the traditional Chinese medicine active monomer composition is matched with the yeast recombinant collagen for use, has obvious treatment effect on skin diseases such as skin barrier function damage, dermatitis, eczema, facial acne and the like, has definite curative effect, makes up the defects of the traditional dressing, has simple preparation process and is suitable for industrial mass production. The invention is safe, has no side effect and can be used for a long time.
Drawings
Fig. 1 is an experimental diagram of the animal acne model curative effect of the second experiment of the invention.
FIG. 2 is the appearance of each major organ of mice in the damaged skin group and the damaged skin control group of experiment three of the present invention.
FIG. 3 is the appearance of each major organ of mice in the normal skin group of experiment three of the present invention.
Detailed Description
The present invention is further described in detail below with reference to specific examples, which are provided for illustration only and are not intended to limit the scope of the present invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1
(1) Weighing 10 parts of propylene glycol, heating to 50 ℃, slowly adding 6 parts of the traditional Chinese medicine active monomer composition, stirring until the traditional Chinese medicine active monomer composition is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 1 part of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 8 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 8 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 1 part of phosphate buffer solution, adjusting the pH value to 6, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 3 parts of ferulic acid, 4 parts of angelone and 3 parts of poria cocos.
Example 2
(1) Weighing 10 parts of propylene glycol, heating to 50 ℃, slowly adding 6 parts of the traditional Chinese medicine active monomer composition, stirring until the traditional Chinese medicine active monomer composition is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 3 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 12 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 12 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 7, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 2 parts of ligustilide, 1 part of ferulic acid, 4 parts of angelone and 4 parts of poria cocos.
Example 3
(1) Weighing 13 parts of propylene glycol, heating to 50 ℃, slowly adding 8 parts of the traditional Chinese medicine active monomer composition, stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 4 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 13 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 12 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 6, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 4 parts of ferulic acid, 6 parts of angelone and 3 parts of poria cocos.
Example 4
(1) Weighing 14 parts of propylene glycol, heating to 50 ℃, slowly adding 7 parts of the traditional Chinese medicine active monomer composition, stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 5 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 14 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 13 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 1 part of phosphate buffer solution, adjusting the pH value to 8, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 3 parts of ligustilide, 1 part of ferulic acid, 6 parts of angelone and 6 parts of poria cocos.
Example 5
(1) Weighing 20 parts of propylene glycol, heating to 50 ℃, slowly adding 8 parts of the traditional Chinese medicine active monomer composition, stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And (3) adding 9 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 15 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 15 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 7, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 5 parts of ferulic acid, 7 parts of angelone and 8 parts of poria cocos.
Example 6
(1) Weighing 15 parts of propylene glycol, heating to 50 ℃, slowly adding 7 parts of the traditional Chinese medicine active monomer composition, stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 6 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 10 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 12 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 7, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 6 parts of ferulic acid, 5 parts of angelone and 8 parts of poria cocos.
Example 7
(1) Weighing 16 parts of propylene glycol, heating to 50 ℃, slowly adding 9 parts of the traditional Chinese medicine active monomer composition, stirring until the traditional Chinese medicine active monomer composition is completely dissolved, and standing for 2 hours to obtain a mixed solution A. (1) Weighing 16 parts of propylene glycol, heating to 50 ℃, and slowly adding 2 parts of ligustilide, 3 parts of ferulic acid, 7 parts of angelone and 4 parts of poria in sequence. Stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 6 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 12 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 10 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 8, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 2 parts of ligustilide, 1 part of ferulic acid, 7 parts of angelone and 4 parts of poria cocos.
Example 8
(1) Weighing 12 parts of propylene glycol, heating to 50 ℃, slowly adding 10 parts of the traditional Chinese medicine active monomer composition, stirring until the traditional Chinese medicine active monomer composition is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 5 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 12 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 13 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 1 part of phosphate buffer solution, adjusting the pH value to 7, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 3 parts of ligustilide, 1 part of ferulic acid, 5 parts of angelone and 6 parts of poria cocos.
Example 9
(1) Weighing 15 parts of propylene glycol, heating to 50 ℃, slowly adding 8 parts of the traditional Chinese medicine active monomer composition, stirring until the mixture is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 5 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 12 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 13 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 2 parts of phosphate buffer solution, adjusting the pH value to 6, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 6 parts of ferulic acid, 4 parts of angelone and 4 parts of poria cocos.
Example 10
(1) Weighing 15 parts of propylene glycol, heating to 50 ℃, slowly adding 9 parts of the traditional Chinese medicine active monomer composition, stirring until the traditional Chinese medicine active monomer composition is completely dissolved, and standing for 2 hours to obtain a mixed solution A.
(2) And adding 6 parts of sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing 13 parts of yeast recombinant collagen, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding 12 parts of glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding 1 part of phosphate buffer solution, adjusting the pH value to 8, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing for treating the skin diseases.
The weight ratio of the raw materials of the Chinese medicinal active monomer composition in the step (1) is 1 part of ligustilide, 6 parts of ferulic acid, 7 parts of angelone and 8 parts of poria cocos.
The liquid dressing prepared in example 10 was investigated for efficacy and safety, and the following experiments were conducted:
first experiment, clinical curative effect experiment
The experimental principle is as follows: clindamycin hydrochloride ointment is commonly used for treating skin diseases such as dermatitis, acne and the like clinically, and the treatment effect and the advantages of the liquid auxiliary materials can be obtained by comparing the clinical curative effect of the liquid dressing prepared in the example 10 with the clinical curative effect of the clindamycin hydrochloride ointment.
Case selection: 20 clinically recorded patients with skin barrier function impairment, dermatitis and eczema, facial acne were randomly drawn and randomized into two groups: group A and group B.
The treatment method comprises the following steps: the A group is prepared from clindamycin hydrochloride ointment by cleaning face before each application, applying on corresponding part, keeping for 20min, washing with clear water, and continuously applying for 3 weeks 2 times per day. Group B the liquid dressing prepared in example 10 was used, and the face was cleaned before each use, applied to the corresponding area, kept for 20min, washed with clean water, and applied 2 times a day for 3 weeks.
Grading standard: the deterioration score is 0, the no effect score is 1, the mild improvement score is 2-3, the moderate improvement score is 4-5, and the obvious improvement score is 6-7.
The statistical method comprises the following steps: data were analyzed using SPSS21.0 statistical software. Effect scoring was performed without a week interval during treatment, and analysis was performed using a repeated measures variance design.
As a result: gender, age difference of the two groups before treatment were not statistically significant; scores of both groups after use were increased compared to those before treatment, and the score of group B was increased more significantly than that of group A, as shown in Table 1.
TABLE 1 comparison of symptom scores before and after treatment in two groups
Note: p < 0.05 compared to pre-treatment;
and (4) conclusion: the liquid adjuvant prepared in example 10 has more significant curative effect on skin diseases than clindamycin hydrochloride ointment.
Experiment two, animal acne model curative effect experiment
The experimental principle is as follows: the therapeutic effect of the liquid dressing prepared in example 10 can be obtained by molding a rabbit, applying the liquid dressing prepared in example 10, and observing the recovery condition of the rabbit.
The experimental method comprises the following steps: culturing and enriching acne bacillus (P.acnes), selecting single colony, pure culturing, and correcting to about 9 × 10 with sterilized normal saline after 7 days 7 CFU/ML. 5 healthy common-grade male rabbits are selected, coal tar is used for damaging the skin of rabbit ears, and the corrected bacterial liquid is injected subcutaneously at the skin damage part of the rabbit ears, wherein each side of the bacterial liquid is about 60 mu L. After 7 days, randomly taking 1 rabbit for skin biopsy, and dividing the rabbits into 2 groups after successful molding is proved, wherein one group is wiped with normal saline every day for 1 time every day for 18 days; the other group was applied with the liquid dressing prepared in example 10 daily 1 time per day for 30min for 18 days.
The experimental results are as follows: after the rabbit ears are injured by coal tar, the rabbit ears are locally thickened and hardened, pores become large, black angle plugs can be seen, and local edema is obvious. The hair follicle pores of the rabbit ears coated with the liquid dressing of example 10 were reduced, the local follicular keratotic plugs and the acne were reduced, and the rabbit ears coated with physiological saline had no significant change from the rabbit ears before treatment. See fig. 1.
And (4) conclusion: the liquid dressing prepared in example 10 has a therapeutic effect on animal skin diseases such as acne.
Experiment III, safety experiment
The experimental principle is as follows: the liquid dressing prepared in example 10 was applied to intact and damaged skin of a mouse, and after a period of time, various indices of the mouse were observed, so that the safety of the liquid dressing could be judged.
The experimental method comprises the following steps: 60 healthy mice were randomly divided into a damaged skin group, a damaged skin control group, and a normal skin group, and 20 mice were administered to each group. The damaged skin group and the control group were subjected to a damage using sandpaper to form a wound surface of 3X 3cm, and the liquid vehicle prepared in example 10 was applied to both the damaged skin group and the normal skin group at a rate of 5g/cm 2 The wound surface is smeared, and the control group of animals are smeared with the same amount of normal saline for 14 days.
The experimental results are as follows: no mouse died in 14 days, and the mouse generally had no abnormality in activity, weight, respiration, skin and eyes, indicating that the liquid dressing prepared in example 10 has no toxicity, and the appearance of each organ is normal after the mouse is dissected. See fig. 2 and 3.
And (4) conclusion: the liquid dressing prepared in example 10 is non-toxic for long term use.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (6)
1. The liquid dressing of the yeast recombinant collagen is characterized by comprising the following raw and auxiliary materials in parts by weight: 8-15 parts of yeast recombinant collagen, 6-10 parts of a traditional Chinese medicine active monomer composition, 1-9 parts of sodium hyaluronate, 8-15 parts of glycerol, 10-20 parts of propylene glycol, 1-2 parts of phosphate buffer solution and the balance of purified water.
2. The yeast recombinant collagen liquid dressing as claimed in claim 1, wherein the Chinese medicinal active monomer composition comprises the following raw materials by weight: 1-3 parts of ligustilide, 1-6 parts of ferulic acid, 4-7 parts of angelone and 3-8 parts of poria cocos.
3. The liquid dressing of yeast recombinant collagen according to claim 1, wherein the preparation method of the liquid dressing comprises:
(1) weighing propylene glycol, heating to 50 deg.C, sequentially and slowly adding Chinese medicinal active monomer, stirring to dissolve completely, and standing for 2 hr to obtain mixed solution A.
(2) And adding sodium hyaluronate into purified water, stirring until the sodium hyaluronate is completely dissolved, and standing for 3 hours to obtain a sodium hyaluronate solution.
(3) Weighing yeast recombinant collagen according to a proportion, adding the yeast recombinant collagen into a sodium hyaluronate solution, stirring until the yeast recombinant collagen is completely dissolved, adding glycerol, continuously stirring for 45min, and standing for 1h at a constant temperature of 20 ℃ to obtain a mixed solution B.
(4) And slowly adding the mixed solution A into the mixed solution B, uniformly stirring, adding purified water to the total amount of the solution, adding a phosphate buffer solution to adjust the pH value, and uniformly stirring to obtain the yeast recombinant collagen liquid dressing.
4. The method for preparing the yeast recombinant collagen liquid dressing as claimed in claim 3, wherein: in the step (4), adding a phosphate buffer solution to adjust the pH value to 6-8.
5. Use of the yeast recombinant collagen liquid dressing obtained according to claim 1 in the adjuvant treatment of skin diseases.
6. Use according to claim 5, characterized in that the skin diseases are disorders of impaired skin barrier function, dermatitis and eczema, facial acne, etc.
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| CN101091890A (en) * | 2007-07-26 | 2007-12-26 | 沈阳药科大学 | Composite type emulsifier, and emulsion prepared by using the emulsifier, and preparation method |
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| CN107753389A (en) * | 2016-11-30 | 2018-03-06 | 江苏创铭医疗器械有限公司 | A kind of recombined collagen foaming pectin facial mask and preparation method thereof |
| CN112190503A (en) * | 2020-11-11 | 2021-01-08 | 华熙生物科技股份有限公司 | Hyaluronic acid composition with penetration promoting effect, preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101091890A (en) * | 2007-07-26 | 2007-12-26 | 沈阳药科大学 | Composite type emulsifier, and emulsion prepared by using the emulsifier, and preparation method |
| CN104667335A (en) * | 2015-01-27 | 2015-06-03 | 山西锦波生物医药股份有限公司 | Recombinant human-derived collagen product for skin barrier function or haemorrhoids and preparation method |
| CN107753389A (en) * | 2016-11-30 | 2018-03-06 | 江苏创铭医疗器械有限公司 | A kind of recombined collagen foaming pectin facial mask and preparation method thereof |
| CN112190503A (en) * | 2020-11-11 | 2021-01-08 | 华熙生物科技股份有限公司 | Hyaluronic acid composition with penetration promoting effect, preparation method and application thereof |
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