CN116999359A - Anti-hair loss stock solution containing transdermal recombinant collagen and preparation method thereof - Google Patents
Anti-hair loss stock solution containing transdermal recombinant collagen and preparation method thereof Download PDFInfo
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- CN116999359A CN116999359A CN202311025794.2A CN202311025794A CN116999359A CN 116999359 A CN116999359 A CN 116999359A CN 202311025794 A CN202311025794 A CN 202311025794A CN 116999359 A CN116999359 A CN 116999359A
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- 108010035532 Collagen Proteins 0.000 title claims abstract description 93
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- 229920001436 collagen Polymers 0.000 title claims abstract description 92
- 230000003656 anti-hair-loss Effects 0.000 title claims abstract description 24
- 239000011550 stock solution Substances 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 244
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 64
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- FHKSXSQHXQEMOK-UHFFFAOYSA-N hexane-1,2-diol Chemical compound CCCCC(O)CO FHKSXSQHXQEMOK-UHFFFAOYSA-N 0.000 claims abstract description 30
- WCVRQHFDJLLWFE-UHFFFAOYSA-N pentane-1,2-diol Chemical compound CCCC(O)CO WCVRQHFDJLLWFE-UHFFFAOYSA-N 0.000 claims abstract description 29
- TXFPEBPIARQUIG-UHFFFAOYSA-N 4'-hydroxyacetophenone Chemical compound CC(=O)C1=CC=C(O)C=C1 TXFPEBPIARQUIG-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229920002385 Sodium hyaluronate Polymers 0.000 claims abstract description 10
- CBTVGIZVANVGBH-UHFFFAOYSA-N aminomethyl propanol Chemical compound CC(C)(N)CO CBTVGIZVANVGBH-UHFFFAOYSA-N 0.000 claims abstract description 10
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- 229940010747 sodium hyaluronate Drugs 0.000 claims abstract description 10
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 claims abstract description 10
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- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
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Classifications
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- 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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
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- 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
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- 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
- A61K8/55—Phosphorus compounds
- A61K8/553—Phospholipids, e.g. lecithin
-
- 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
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
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- 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
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- 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
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- 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
- A61K8/81—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions involving only carbon-to-carbon unsaturated bonds
- A61K8/8141—Compositions of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Compositions of derivatives of such polymers
- A61K8/8147—Homopolymers or copolymers of acids; Metal or ammonium salts thereof, e.g. crotonic acid, (meth)acrylic acid; Compositions of derivatives of such polymers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q7/00—Preparations for affecting hair growth
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- 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
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
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- Pharmacology & Pharmacy (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses an anti-hair loss stock solution containing transdermal recombinant collagen and a preparation method thereof. The anti-hair loss stock solution comprises the following components in percentage by mass: 0.5-2% of recombinant collagen glycerol body, 2-10% of glycerol, 0.5-5% of 1, 2-hexanediol, 0.5-6% of p-hydroxyacetophenone, 0.1-0.7% of sodium hyaluronate, 0.5-5% of 1, 2-pentanediol, 0.05-2% of aminomethylpropanol, 0.05-2% of plant sensitivity-relieving agent SGS, 0.1-2% of carbomer U21 and the balance of water. The invention utilizes the glycerin encapsulation technology to encapsulate the high-activity recombinant collagen in the glycerin vesicle, improves the transdermal property of the recombinant collagen, and combines the transdermal recombinant collagen serving as an active ingredient with other raw materials to form the alopecia-preventing stock solution, which not only has the effects of moisturizing and locking water to skin, but also has the effect of preventing alopecia.
Description
Technical Field
The invention belongs to the field of cosmetic preparations, and relates to an anti-hair loss stock solution containing transdermal recombinant collagen and a preparation method thereof.
Background
In recent years, with the increasing intensity of social competition, people have increased mental pressure, increasingly worsened environment and fashion of hair dyeing and perming, the occurrence proportion of alopecia is in an ascending trend, the trend of younger age is more and more obvious, the alopecia anxiety of young people has become a common phenomenon, and the consumption scale of anti-hair-loss and hair-growing products is continuously enlarged.
The active ingredients added in the existing anti-hair loss products mainly comprise some traditional Chinese medicine components or organic macromolecules. For example, CN101904908A discloses a traditional Chinese medicine composition for preventing alopecia, which comprises extracts of the following traditional Chinese medicinal materials: ginseng, kuh-seng, cowherb seed, garden balsam stem, mulberry leaf, liquorice, rhubarb and safflower. CN104814884a discloses a shampoo for preventing hair loss and promoting hair growth, which uses ultra-high molecular weight (70-300 tens of thousands) gamma-polyglutamic acid as an active ingredient, which can effectively inhibit the activity of 5α -reductase, thereby treating hair loss and promoting hair growth. The shampoo contains collagen, but is mainly used as a nutritional ingredient.
The recombinant collagen disclosed in CN102443057B is a recombinant human collagen which is based on the tripeptide repeat sequence characteristics of human type III collagen alpha 1 chain collagen domain Gly-X-Y (glycine-X-Y), is designed and artificially synthesized into a segment of human collagen gene monomer and expressed by using Pichia pastoris, and has unique chemical structure and performance superior to that of animal collagen. The recombinant collagen is mainly used as an anti-aging and nutritional ingredient applied to skin care products or used as a basic raw material applied to the preparation of medical instruments such as dressing and the like. There are no reports of its application in hair care products.
Disclosure of Invention
The invention aims to provide an anti-hair loss stock solution containing transdermal recombinant collagen and a preparation method thereof. The hair loss preventing stock solution takes transdermal recombinant collagen with excellent skin affinity and histocompatibility as raw material, and has hair loss preventing effect.
The technical scheme for realizing the purpose of the invention is as follows:
the anti-hair loss stock solution containing transdermal recombinant collagen comprises the following components in percentage by mass: 0.5 to 2 percent of recombinant collagen glycerol body, 2 to 10 percent of glycerol, 0.5 to 5 percent of 1, 2-hexanediol, 0.5 to 6 percent of p-hydroxyacetophenone, 0.1 to 0.7 percent of sodium hyaluronate, 0.5 to 5 percent of 1, 2-pentanediol, 0.05 to 2 percent of aminomethylpropanol, 0.05 to 2 percent of plant sensitivity agent SGS, 210.1 to 2 percent of carbomer U and the balance of water; the recombinant collagen glycerol body is composed of the following components in parts by weight as 100 parts: 2.7-3.7 parts of recombinant collagen, 5-15 parts of glycerol, 3.0-4.0 parts of L-alpha-phosphatidylcholine, 0.55-0.75 part of emulsion regulator cetyl alcohol, 2 parts of preservative 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.05-0.10 part of thickener xanthan gum, 0.1 part of sodium chloride and 75.05-84.9 parts of purified water, wherein the recombinant collagen is produced by fermenting Pichia pastoris with the preservation number of CGMCC No. 5021.
The recombinant collagen of the invention is fully disclosed in Chinese patent CN 102443057B.
Preferably, the recombinant collagen glycerol body is composed of the following components in parts by weight based on 100 parts by weight: 3.2 parts of recombinant collagen, 10 parts of glycerol, 3 parts of L-alpha-phosphatidylcholine, 0.65 part of emulsion regulator cetyl alcohol, 2 parts of preservative 1, 2-pentanediol and 0.5 part of 1, 2-hexanediol, 0.08 part of thickener xanthan gum, 0.1 part of sodium chloride and 80.47 parts of purified water.
Preferably, the average particle size of the recombinant collagen glycerome vesicles is 150-300 nm.
The preparation method of the recombinant collagen glyceride adopts an aqueous phase precipitation method and comprises the following steps:
(1) Dissolving L-alpha-phosphatidylcholine in ethanol with the mass being 5 times of that of the L-alpha-phosphatidylcholine under the water bath condition of 65-85 ℃ according to the proportion, then adding an emulsion regulator cetyl alcohol, and uniformly stirring to obtain an organic solvent system;
(2) Completely dissolving glycerol, recombinant collagen, thickener xanthan gum, preservative 1, 2-pentanediol, 1, 2-hexanediol, sodium chloride and purified water in water bath at 50-60 ℃ and uniformly mixing to obtain a water phase system;
(3) And (3) dropwise adding the organic solvent system into the water phase system, simultaneously carrying out ultrasonic treatment to evaporate ethanol to obtain a glycerol body, and then filtering through a 0.45 mu m hydrophilic filter membrane to obtain the uniform and stable recombinant collagen glycerol body.
Preferably, the anti-hair loss stock solution containing transdermal recombinant collagen comprises the following components in percentage by mass: 1% of recombinant collagen glycerol, 2% of glycerol, 0.5% of 1, 2-hexanediol, 0.5% of p-hydroxyacetophenone, 0.3% of sodium hyaluronate, 1, 2-pentanediol, 0.1% of aminomethylpropanol, 0.5% of plant sensitivity-relieving agent SGS, 210.2% of carbomer U and the balance of water.
The preparation method of the anti-hair loss stock solution containing transdermal recombinant collagen comprises the following steps:
sequentially adding sodium hyaluronate, carbomer U21, glycerol and p-hydroxyacetophenone into water according to a proportion, heating to 65+/-5 ℃, uniformly stirring, then cooling to below 45 ℃, adding recombinant collagen glycerol, 1, 2-pentanediol, aminomethylpropanol, plant sensitivity agent SGS and 1, 2-hexanediol, uniformly stirring until the components are completely dissolved, and obtaining the anti-hair loss stock solution containing transdermal recombinant collagen.
Compared with the prior art, the invention has the following advantages:
(1) The invention takes the recombinant collagen with excellent skin affinity and tissue compatibility as a basic component, and utilizes the glycerin body wrapping technology to wrap the high-activity recombinant collagen in the glycerin body vesicle, so that the glycerin body vesicle has the characteristics of high stability, controllable slow release, safety and mildness, and meanwhile, the water solubility of the recombinant collagen glycerin body is good, the transdermal property of the recombinant collagen is improved, and the recombinant collagen is accelerated to permeate through the skin epidermis and permeate into the skin basal layer, so that the recombinant collagen is easy to be absorbed by a human body.
(2) According to the invention, components such as glycerol, 1, 2-hexanediol and the like are added, the glycerol, the 1, 2-hexanediol, the 1, 2-pentanediol and the sodium hyaluronate are all used as moisturizers, the p-hydroxyacetophenone is used as an antioxidant, the aminomethylpropanol is used as a pH regulator, the plant sensitivity-relieving agent SGS is used as a skin conditioner, the recombinant collagen glycerol body is used as the skin conditioner, and the formed anti-hair loss stock solution has the effects of moisturizing and water locking on skin, and also can promote hair regeneration, nourish scalp and improve living environment, and has the anti-hair loss effect.
Detailed Description
The invention will be further described in detail with reference to specific examples.
The recombinant collagen used in the examples below was purchased from Jiangsu Jiangshan poly-source biotechnology limited.
In the following examples and comparative examples, the method for measuring subcutaneous penetration rate is as follows:
ex vivo skin test: 100. Mu.g/cm by micropipette 2 The products (based on recombinant collagen content) were each smeared onto ex vivo pig skin and the skin was exposed in a diffusion chamber for 7 hours, the temperature being maintained at 35.+ -. 1 ℃. After 7 hours of exposure, the skin was removed. The stratum corneum was removed by tape and the epidermal basal layer was separated. All skin sites and tapes were immersed in 2.5ml of 50% aqueous methanol (v/v) solution. The samples were then sonicated for 30 minutes to extract collagen from the skin, which was then quantitatively detected by HPLC.
Example 1
The recombinant collagen glycerol body of this example was prepared by the following steps:
3.2 parts of recombinant collagen, 10 parts of glycerol, 3 parts of L-alpha-phosphatidylcholine, 0.65 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.08 part of xanthan gum, 0.1 part of sodium chloride and 80.47 parts of purified water are respectively weighed according to parts by weight. Dissolving L-alpha-phosphatidylcholine in ethanol with the mass of 5 times under the water bath condition of 75 ℃, then adding cetyl alcohol, and uniformly stirring to obtain an organic solvent system; completely dissolving glycerol, recombinant collagen, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water in a water bath at 55 ℃, and uniformly mixing to obtain a water phase system; and (3) dropwise adding the organic solvent system into the water phase system, simultaneously carrying out ultrasonic treatment to evaporate ethanol to obtain a glycerol body, and then filtering through a 0.45 mu m hydrophilic filter membrane to finally obtain the uniform and stable recombinant collagen glycerol body.
Example 2
The recombinant collagen glycerol body of this example was prepared by the following steps:
2.7 parts of recombinant collagen, 5 parts of glycerol, 4 parts of L-alpha-phosphatidylcholine, 0.75 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.05 part of xanthan gum, 0.1 part of sodium chloride and 84.9 parts of purified water are respectively weighed according to parts by weight. Dissolving L-alpha-phosphatidylcholine in ethanol with the mass of 5 times under the water bath condition of 85 ℃, adding cetyl alcohol, and uniformly stirring to obtain an organic solvent system; completely dissolving glycerol, recombinant collagen, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water in water bath at 60 ℃, and uniformly mixing to obtain a water phase system; and (3) dropwise adding the organic solvent system into the water phase system, simultaneously carrying out ultrasonic treatment to evaporate ethanol to obtain a glycerol body, and then filtering through a 0.45 mu m hydrophilic filter membrane to finally obtain the uniform and stable recombinant collagen glycerol body.
Example 3
The recombinant collagen glycerol body of this example was prepared by the following steps:
3.7 parts of recombinant collagen, 15 parts of glycerol, 3.0 parts of L-alpha-phosphatidylcholine, 0.55 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.1 part of xanthan gum, 0.1 part of sodium chloride and 75.05 parts of purified water are respectively weighed according to parts by weight. Dissolving L-alpha-phosphatidylcholine in ethanol with the mass of 5 times under the water bath condition of 65 ℃, then adding cetyl alcohol, and uniformly stirring to obtain an organic solvent system; completely dissolving glycerol, recombinant collagen, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water in a water bath at 50 ℃, and uniformly mixing to obtain a water phase system; and (3) dropwise adding the organic solvent system into the water phase system, simultaneously carrying out ultrasonic treatment to evaporate ethanol to obtain a glycerol body, and then filtering through a 0.45 mu m hydrophilic filter membrane to finally obtain the uniform and stable recombinant collagen glycerol body.
Comparative example 1
The comparative example was substantially identical to the composition and preparation method of example 1, except that the two dissolution temperatures were different, specifically, L- α -phosphatidylcholine was dissolved in 5 times mass of ethanol under the water bath condition of 95℃and glycerin, recombinant collagen, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water were completely dissolved under the water bath condition of 45 ℃.
Comparative example 2
The comparative example was substantially identical to the composition and preparation method of example 2, except that the two dissolution temperatures were different, specifically, L- α -phosphatidylcholine was dissolved in 5 times mass of ethanol under the water bath condition of 50℃and glycerin, recombinant collagen, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water were completely dissolved under the water bath condition of 80 ℃.
Comparative example 3
The comparative example does not add glycerol and comprises the following components: 3.7 parts of recombinant collagen, 3.0 parts of L-alpha-phosphatidylcholine, 0.55 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.1 part of xanthan gum, 0.1 part of sodium chloride and 90.05 parts of purified water, and the preparation method is the same as that of example 1.
Comparative example 4
This comparative example is substantially identical to the composition and preparation method of example 1, except that L-alpha-phosphatidylcholine is replaced with phosphatidylglycerol.
Comparative example 5
The method for preparing the recombinant collagen glycerol body by adopting the thin film evaporation method comprises the following specific steps:
3.2 parts of recombinant collagen, 10 parts of glycerol, 3 parts of L-alpha-phosphatidylcholine, 0.65 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.08 part of xanthan gum, 0.1 part of sodium chloride and 80.47 parts of purified water are respectively weighed according to parts by weight. Dissolving L-alpha-phosphatidylcholine and glycerol in ethanol with the mass of 5 times under the water bath condition of 75 ℃ and rotationally evaporating to obtain a uniform glycerolipid membrane; completely dissolving recombinant collagen, cetyl alcohol, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water in a water bath at 55 ℃, and uniformly mixing to obtain a mixed liquid; adding the mixed liquid into a glycerol body lipid membrane, carrying out ultrasonic crushing, uniformly mixing, homogenizing, and then filtering through a 0.45 mu m hydrophilic filter membrane to finally obtain a glycerol suspension, namely the recombinant collagen glycerol body.
Comparative example 6
The preparation method of the recombinant collagen glycerol body by adopting the lipid fusion method comprises the following specific steps:
3.2 parts of recombinant collagen, 10 parts of glycerol, 3 parts of L-alpha-phosphatidylcholine, 0.65 part of cetyl alcohol, 2 parts of 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.08 part of xanthan gum, 0.1 part of sodium chloride and 80.47 parts of purified water are respectively weighed according to parts by weight. Dissolving L-alpha-phosphatidylcholine and glycerol in ethanol with the mass of 5 times under the water bath condition of 95 ℃ and rotationally evaporating to obtain a uniform glycerolipid membrane; completely dissolving recombinant collagen, cetyl alcohol, 1, 2-pentanediol, 1, 2-hexanediol, xanthan gum, sodium chloride and purified water in a water bath at 45 ℃, and uniformly mixing to obtain a mixed liquid; adding the mixed liquid into a glycerol body lipid membrane, carrying out ultrasonic crushing, uniformly mixing, homogenizing, and then filtering through a 0.45 mu m hydrophilic filter membrane to finally obtain a glycerol suspension, namely the recombinant collagen glycerol body.
Table 1 composition of the components of each of examples and comparative examples
Table 2 Performance data of samples prepared in each experimental example
As can be seen from comparison of comparative examples 1,2 and examples, when the water bath temperature of the organic solvent system is too high or too low and the water bath temperature of the water-soluble system is too low or too high, the appearance of the finally prepared product cannot form uniform emulsion, the particle size of the product is larger, the encapsulation efficiency is lower, the collagen degradation rate in the aqueous solution is faster, and the subcutaneous permeability is obviously weakened, and the results show that the water bath temperature is suitable for the specific recombinant collagen glycerol body solution of the invention, and is more favorable for the formation of regular particle sizes of liposome, and the water bath temperature has different effects on the product appearance encapsulation efficiency, the stability of the recombinant collagen in the aqueous solution and the collagen transdermal property, so that the preparation of the recombinant collagen glycerol body requires proper process conditions.
Comparative example 3 lacks glycerol in the glycerol body skeleton component, comparative example 4 replaces phospholipid in the glycerol body skeleton component and L-alpha-phosphatidylcholine with phosphatidylglycerol, the final products of comparative examples 3 and 4 have large particle size and low encapsulation efficiency, and collagen in the products has high degradation speed in aqueous solution and poorer transdermal property. The results show that the glycerol body framework component can ensure the stability and the transdermal property of the product, the glycerol body framework component cannot be absent, and the phospholipid in the framework component is L-alpha-phosphatidylcholine, so that the stability and the transdermal property of the glycerol body can be obviously improved.
Compared with example 1, comparative examples 5 and 6 respectively adopt a thin film evaporation method and a lipid fusion method to synthesize collagen glycerol, the particle size of the finally prepared product is large, the encapsulation efficiency is low, the degradation speed of collagen in the product in aqueous solution is too high, and the transdermal property is poorer. The above results indicate that the preparation of recombinant collagen glycerol bodies is more suitable by an aqueous precipitation method.
Example 4
1. Experimental grouping
The experiment is divided into two groups of test products and control products, in order to ensure the true accuracy of the test, the same formula and the same batch of stock solution are used as a matrix, 1% of recombinant collagen glycerol body is added on the matrix in the test group, and 1% of purified water is added in the control product.
2. Experimental sample
(1) Composition of the components of the test product: 1% of recombinant collagen glycerol, 2% of glycerol, 0.5% of 1, 2-hexanediol, 0.5% of p-hydroxyacetophenone, 0.3% of sodium hyaluronate, 1, 2-pentanediol, 0.1% of aminomethylpropanol, 0.5% of plant sensitivity-relieving agent SGS, 210.2% of carbomer U and the balance of water.
(2) Composition of the control product: 2% of glycerol, 0.5% of 1, 2-hexanediol, 0.5% of p-hydroxyacetophenone, 0.3% of sodium hyaluronate, 1% of 1, 2-pentanediol, 0.1% of aminomethylpropanol, 0.5% of plant sensitivity agent SGS, 210.2% of carbomer U and the balance of water.
3. Test purpose: the anti-hair loss efficacy of the recombinant collagen glycerol body was tested.
4. Eluting the product: as with the control product, both the test and control groups used the control product during the elution period.
5. The subject: total 61, men 3, women 58, ages 21 to 60, mean ages 43.98±9.36, and met the subject's volunteer inclusion criteria.
6. Instrument apparatus: professional digital cameras: the pixel is not less than 1500 ten thousand, and parameters including aperture size, photosensitivity (ISO), focal length and the like are kept consistent in the whole test process; image capturing support: the head position of the subject can be fixed, the head top is kept vertical to the camera lens, and all hair photos are shot by matching with a professional digital camera by taking the head top as the center; skin mirror: LED lamp light source with wavelength of 450-750 nm, color temperature of 6500-8000K, illuminance not lower than 540lx, amplification factor not less than 20 times, detection diameter not less than 1.0cm or area not less than 0.8cm 2 The method comprises the steps of carrying out a first treatment on the surface of the Comb: the comb teeth have moderate density (the tooth space is 0.9-1.1 mm), the length of the comb teeth is 2.0-3.0 cm, the length of the comb is not less than 10cm (no comb handle is contained), the comb with the same specification and material is required to be used in the whole test process, and the comb is required to be disinfected after each use by referring to related requirements in the medical institution disinfection technical Specification (WS-T367-2012).
7. Environmental requirements: test results observations should be made in an environment with a temperature of 21±1 ℃ and a relative humidity of 50±10%rh, visual assessment should be made under constant illumination conditions (fluorescent tubes or LED illumination with a color temperature of 5500 to 6500K), and subjects should be allowed to evaluate and test after at least 30 minutes of adaptation under such environmental conditions.
8. The test method comprises the following steps: the test was carried out according to the specific requirements of cosmetic safety Specification (2015 edition). Before the group is put into the group, the subjects are inquired about a series of problems about disease history, health condition and the like according to the inclusion and exclusion criteria and the like, and meanwhile, the number of alopecia is counted by adopting a 60-time combing method and recorded. Qualified subjects undergo a 2-week washout period, 60 hair combing methods are performed again after the washout period is finished, the hair loss count is still greater than 10, the subjects who enter the formal test can enter the formal test, the subjects who enter the formal test are divided into a test product group and a control product group according to a layering random method, and balance of important factors (sex, age, hair length, hair loss severity and the like) possibly influencing test results is ensured. The staff dispenses test products and control products according to a random table, and guides the use of the products to the test subjects according to the use instructions, so that the test subjects can be ensured to use the test subjects correctly in the test period. The test substance is used for at least 12 weeks, and the test period requires the test subject to record the use time and any discomfort and adverse reaction symptoms in the use process; performing evaluation of hair basic values before using products on selected subjects, including hair loss counting, hair density evaluation and image shooting, and recording; the same evaluation and testing was again performed 4, 8, 12 weeks after product use.
9. Data statistics: statistical analysis of the data was performed using statistical analysis software. The metering data are expressed as: the mean value is +/-standard deviation, normal distribution inspection is carried out, normal distribution requirements are met, paired t inspection is adopted for comparison before and after the mean value is self, and otherwise, two related sample rank sum inspection is adopted; comparing the grade data before and after use, and adopting two related sample ranks and tests; comparison between test product and control group used independent sample t-test or rank-sum test. The above statistical analysis was a two-tailed test with a significance level of α=0.05.
10. Conclusion judgment: according to the data analysis result, any visit time point during the test period is not significantly increased in the hair loss count before and after using the test product or the difference in the hair loss count before and after using the test product (the hair loss count at a certain visit time point after using the product-the hair loss count before using the product) is significantly lower than that of the control group (p < 0.5), the test product is determined to have the anti-hair loss effect, otherwise, the test product is determined to have no anti-hair loss effect.
11. Data analysis of test results:
(1) The specific test results of the metering data are shown in tables 3, 4, 5, 6 and attached table 1.
(2) The specific test results of the grade data are shown in Table 7, table 8 and Table 2.
TABLE 3 statistical analysis of alopecia counts between time points before and after product use
TABLE 4 statistical analysis of alopecia count for test product area versus control product area at various time points
* The statistical method comprises the following steps: analysis was performed using t-test or rank sum test methods, with a test level of a=0.05.
* The significance labeling method comprises the following steps: ("n.s" means no statistical difference, p >0.05; s' means significant difference, p < 0.05).
Table 5 statistical analysis of local hair density between time points before and after product use
Table 6 results of statistical analysis of local hair density in test product areas versus control product areas at various time points
* The statistical method comprises the following steps: analysis was performed using t-test or rank sum test methods, with a test level of a=0.05.
* The significance labeling method comprises the following steps: ("n.s" means no statistical difference, p >0.05; s' means significant difference, p < 0.05).
TABLE 7 statistical analysis of overall hair density between time points before and after product use
Table 8 statistical analysis of overall hair density for test product area versus control product area at various time points
* The statistical method comprises the following steps: analysis was performed using t-test or rank sum test methods, with a test level of a=0.05.
* The significance labeling method comprises the following steps: ("n.s" means no statistical difference, p >0.05; s' means significant difference, p < 0.05).
12. Conclusion of the test
(1) After the test product is used, the alopecia count of the test product group is obviously reduced (p < 0.05) at each return visit time point compared with that before the test product is used; the hair loss count improvement of the test product group was significantly better than that of the control group (p < 0.05) at each return visit time point.
(2) After the test product is used, the local hair density of the test product group is obviously increased (p is less than 0.05) at 4 weeks and 12 weeks compared with the hair density before the test product is used, and is not obviously reduced (p is more than or equal to 0.05) at 8 weeks compared with the hair density before the test product is used; the local hair density of the test product group was not significantly different from that of the control group at each return visit time point (p.gtoreq.0.05).
(3) After the test product is used, the overall hair density of the test product group is not significantly reduced (p is more than or equal to 0.05) at 4 weeks and 8 weeks compared with the prior use; the overall hair density of the test product group was not significantly different from that of the control group at each return visit time point (p.gtoreq.0.05). The result shows that the anti-drop stock solution containing the transdermal recombinant collagen has obvious anti-drop effect.
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Claims (6)
1. The anti-hair loss stock solution containing transdermal recombinant collagen is characterized by comprising the following components in percentage by mass: 0.5-2% of recombinant collagen glycerol body, 2-10% of glycerol, 0.5-5% of 1, 2-hexanediol, 0.5-6% of p-hydroxyacetophenone, 0.1-0.7% of sodium hyaluronate, 0.5-5% of 1, 2-pentanediol, 0.05-2% of aminomethylpropanol, 0.05-2% of plant sensitivity-relieving agent SGS, 0.1-2% of carbomer U21 and the balance of water; the recombinant collagen glycerol body is composed of the following components in parts by weight as 100 parts: 2.7-3.7 parts of recombinant collagen, 5-15 parts of glycerol, 3.0-4.0 parts of L-alpha-phosphatidylcholine, 0.55-0.75 part of emulsion regulator cetyl alcohol, 2 parts of preservative 1, 2-pentanediol, 0.5 part of 1, 2-hexanediol, 0.05-0.10 part of thickener xanthan gum, 0.1 part of sodium chloride and 75.05-84.9 parts of purified water, wherein the recombinant collagen is produced by fermenting Pichia pastoris with the preservation number of CGMCC No. 5021.
2. The anti-hair loss stock solution according to claim 1, wherein the recombinant collagen glycerol body comprises the following components in parts by weight based on 100 parts by weight: 3.2 parts of recombinant collagen, 10 parts of glycerol, 3 parts of L-alpha-phosphatidylcholine, 0.65 part of emulsion regulator cetyl alcohol, 2 parts of preservative 1, 2-pentanediol and 0.5 part of 1, 2-hexanediol, 0.08 part of thickener xanthan gum, 0.1 part of sodium chloride and 80.47 parts of purified water.
3. The anti-hair loss stock solution according to claim 1 or 2, wherein the average particle size of recombinant collagen glycerol vesicles is 150-300 nm.
4. The anti-hair loss stock solution according to claim 1 or 2, wherein the preparation method of the recombinant collagen glycerol body adopts an aqueous phase precipitation method, and comprises the following steps:
(1) Dissolving L-alpha-phosphatidylcholine in ethanol with the mass being 5 times of that of the L-alpha-phosphatidylcholine under the water bath condition of 65-85 ℃ according to the proportion, then adding an emulsion regulator cetyl alcohol, and uniformly stirring to obtain an organic solvent system;
(2) Completely dissolving glycerol, recombinant collagen, thickener xanthan gum, preservative 1, 2-pentanediol, 1, 2-hexanediol, sodium chloride and purified water in water bath at 50-60 ℃ and uniformly mixing to obtain a water phase system;
(3) And (3) dropwise adding the organic solvent system into the water phase system, simultaneously carrying out ultrasonic treatment to evaporate ethanol to obtain a glycerol body, and then filtering through a 0.45 mu m hydrophilic filter membrane to obtain the uniform and stable recombinant collagen glycerol body.
5. The anti-hair loss stock solution according to claim 1, comprising the following components in percentage by mass: 1% of recombinant collagen glycerol, 2% of glycerol, 0.5% of 1, 2-hexanediol, 0.5% of p-hydroxyacetophenone, 0.3% of sodium hyaluronate, 1, 2-pentanediol, 0.1% of aminomethylpropanol, 0.5% of plant sensitivity-relieving agent SGS, 0.2% of carbomer U21 and the balance of water.
6. The method for preparing the anti-hair loss stock solution according to claim 1, comprising the steps of:
sequentially adding sodium hyaluronate, carbomer U21, glycerol and p-hydroxyacetophenone into water according to a proportion, heating to 65+/-5 ℃, uniformly stirring, then cooling to below 45 ℃, adding recombinant collagen glycerol, 1, 2-pentanediol, aminomethylpropanol, plant sensitivity agent SGS and 1, 2-hexanediol, uniformly stirring until the components are completely dissolved, and obtaining the anti-hair loss stock solution containing transdermal recombinant collagen.
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