CN114470171B - Collagen liposome freeze-dried emulsion and preparation method thereof - Google Patents
Collagen liposome freeze-dried emulsion and preparation method thereof Download PDFInfo
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- 239000002502 liposome Substances 0.000 title claims abstract description 82
- 102000008186 Collagen Human genes 0.000 title claims abstract description 78
- 108010035532 Collagen Proteins 0.000 title claims abstract description 78
- 229920001436 collagen Polymers 0.000 title claims abstract description 78
- 239000000839 emulsion Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 238000004945 emulsification Methods 0.000 title description 2
- 239000011259 mixed solution Substances 0.000 claims abstract description 29
- 238000001035 drying Methods 0.000 claims abstract description 25
- 239000000243 solution Substances 0.000 claims abstract description 23
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 22
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 21
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 21
- 229930195725 Mannitol Natural products 0.000 claims abstract description 21
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 21
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims abstract description 21
- 229960004853 betadex Drugs 0.000 claims abstract description 21
- 235000010355 mannitol Nutrition 0.000 claims abstract description 21
- 239000000594 mannitol Substances 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 11
- 235000013336 milk Nutrition 0.000 claims abstract description 11
- 239000008267 milk Substances 0.000 claims abstract description 11
- 210000004080 milk Anatomy 0.000 claims abstract description 11
- 230000008014 freezing Effects 0.000 claims description 23
- 238000003756 stirring Methods 0.000 claims description 22
- 238000007710 freezing Methods 0.000 claims description 17
- 239000008213 purified water Substances 0.000 claims description 16
- 238000001816 cooling Methods 0.000 claims description 10
- 241000235058 Komagataella pastoris Species 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims 1
- 230000000975 bioactive effect Effects 0.000 abstract description 6
- 239000000463 material Substances 0.000 abstract description 6
- 238000004108 freeze drying Methods 0.000 abstract description 4
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 abstract 1
- 230000002349 favourable effect Effects 0.000 abstract 1
- 238000010438 heat treatment Methods 0.000 description 18
- 230000000052 comparative effect Effects 0.000 description 8
- 239000013543 active substance Substances 0.000 description 4
- 150000003904 phospholipids Chemical class 0.000 description 3
- 239000000232 Lipid Bilayer Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 230000008776 intercellular pathway Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000012792 lyophilization process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 210000000434 stratum corneum Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/40—Cyclodextrins; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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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
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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
- A61P17/18—Antioxidants, e.g. antiradicals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/064—Saccharomycetales, e.g. baker's yeast
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Abstract
The invention discloses collagen liposome freeze-dried emulsion and a preparation method thereof. According to the method, mannitol and beta-cyclodextrin are sequentially added into a collagen liposome solution according to a proportion to form a mixed solution, and then a freeze-drying process is adopted to pump out solution solutes to prepare freeze-dried milk. The invention adopts the freeze-drying process to prepare the collagen liposome freeze-dried emulsion, can force the solution to supercool and crystallize, is favorable for improving the drying efficiency under the negative pressure condition, forms a loose structure along with the gradual extraction of water in the solution, has the function of a framework by mannitol, and can surround the bioactive material in the drying process by beta-cyclodextrin, thereby improving the stability of the liposome structure. Under the dry environment, the collagen enzymolysis can be effectively prevented, the stability of the product is improved, and the long-term storage of the product is facilitated.
Description
Technical Field
The invention relates to the technical field of medical instruments, and relates to collagen liposome freeze-dried emulsion and a preparation method thereof.
Background
Liposomes (Liposomes, lips) are miniature vesicles formed by encapsulating an active substance in a bilayer of lipids, wherein the liposomes can encapsulate a water-soluble drug in an internal aqueous vesicle or a lipid-soluble drug in a lipid bilayer, and then directly act on the lipid bilayer of a keratinocyte to improve permeation of the active substance and promote transdermal absorption of the active substance, and the liposomes can penetrate the stratum corneum completely through skin appendages or intercellular pathways to the dermis layer, so that an active substance reservoir is formed and acts on an affected part slowly for a long time. The phospholipid and cholesterol in the liposome are natural components of human body, and have no irritation and toxicity to human body.
The collagen liposome is prepared by wrapping high-activity recombinant collagen (RHC for short) in liposome vesicles by liposome wrapping technology, so that the collagen liposome can quickly penetrate through skin epidermis and completely penetrate into basal layers of the skin, and the effects of resisting aging, repairing and the like are rapidly exerted. However, liposomes are unstable in aqueous environments and are prone to leakage from solution and phospholipid hydrolysis.
Disclosure of Invention
The invention aims to provide collagen liposome freeze-dried milk and a preparation method thereof.
The technical scheme for realizing the purpose of the invention is as follows:
The collagen liposome freeze-dried emulsion comprises the following components in parts by weight based on 100 parts by weight: 10 parts of collagen liposome, 5-10 parts of mannitol, 0.5-2 parts of beta-cyclodextrin and 78-84.5 parts of purified water, and the collagen liposome is prepared by the following steps:
(1) Adding collagen liposome (LLA) into purified water according to a proportion, stirring and dissolving at normal temperature to obtain collagen liposome solution, then sequentially adding mannitol and beta-cyclodextrin, stirring and dissolving at normal temperature to obtain mixed solution;
(2) Placing the mixed solution in a freeze dryer, setting the cooling temperature to be 5+/-1 ℃, pre-cooling for 20-30 min, then starting a pre-freezing procedure, wherein the freezing temperature is-40 to-50 ℃, and the freezing time is 2-3 h;
(3) After the pre-freezing beam, the procedure automatically enters a primary drying stage, the vacuum degree is set to be 0.1-0.5 mbar, the temperature of the plate layer is set to be minus 25-minus 20 ℃, the holding time is 3-4 h, the temperature of the plate layer is increased to minus 15 ℃, the holding time is 2-3 h, the temperature of the plate layer is increased to be 0 ℃, and the holding time is 1-2 h;
(4) After the primary drying is finished, the procedure automatically enters a secondary drying stage, the temperature of the plate layer is set to be raised to 15 ℃, the holding time is 1-1.5 h, the vacuum degree is set to be 0.01mbar, the temperature of the plate layer is raised to 30 ℃, and the holding time is 2-4 h, so that the collagen liposome freeze-dried emulsion is obtained.
The collagen in the collagen liposome is recombinant human collagen, and is produced by fermenting Pichia pastoris with the preservation number of CGMCC No.5021, and the molecular weight of the collagen liposome is 55kDa.
The average grain diameter of the collagen liposome vesicle is 300-320 nm. Compared with the prior art, the invention has the following advantages:
(1) The invention takes collagen liposome (LLA), mannitol and beta-cyclodextrin as raw materials to prepare the collagen liposome freeze-dried emulsion, wherein the mannitol can be used as a framework agent in a liposome freeze-drying process to surround the bioactive material in the drying process to form a porous reticular structure, so that the bioactive material is prevented from being directly exposed in the surrounding environment, the beta-cyclodextrin can be used for surrounding the bioactive material in the drying process, the stability of the liposome vesicle structure is improved, and the encapsulation rate of the product is ensured.
(2) The freeze-dried collagen liposome emulsion is prepared by adopting a freeze-drying process, so that the solution can be forced to be supercooled and crystallized, the drying efficiency is improved under the negative pressure condition, the water in the solution is gradually pumped away, a loose structure is formed in the drying part, and mannitol plays a role of a framework. Under the dry environment, the collagen enzymolysis is effectively prevented, the stability of the product is improved, and the long-term storage of the product is facilitated.
Drawings
FIG. 1 is a schematic diagram of the collagen liposome product prepared in example 1;
FIG. 2 is a schematic diagram of the collagen liposome prepared in comparative example 1.
Detailed Description
The invention is further described in detail below with reference to examples and figures.
Collagen liposomes used in the examples described below were purchased from Jiangsu Jiangshan Seiyuan Biotechnology Co.
Example 1
The collagen liposome lyophilized milk of this example was prepared by the following steps:
10 parts of collagen liposome, 5 parts of mannitol, 0.5 part of beta-cyclodextrin and 84.5 parts of purified water are respectively weighed according to parts by mass. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer at 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, freezing at-40 ℃ for 3h to obtain a frozen product, adjusting the temperature of the plate layer to-20 ℃, setting the vacuum degree to 0.5mbar, keeping for 4h, heating the plate layer to-15 ℃ for 3h, and heating the plate layer to 0 ℃ for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 4h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Example 2
The collagen liposome lyophilized milk of this example was prepared by the following steps:
10 parts of collagen liposome, 10 parts of mannitol, 2 parts of beta-cyclodextrin and 78 parts of purified water are respectively weighed according to parts by mass. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer at 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, freezing at-40 ℃ for 2h to obtain a frozen product, adjusting the temperature of the plate layer to-25 ℃, setting the vacuum degree to 0.1mbar, keeping for 4h, heating the plate layer to-15 ℃ for 3h, and heating the plate layer to 0 ℃ for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 4h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Example 3
The collagen liposome lyophilized milk of this example was prepared by the following steps:
10 parts of collagen liposome, 5 parts of mannitol, 0.5 part of beta-cyclodextrin and 84.5 parts of purified water are respectively weighed according to parts by mass. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer at 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, freezing at-40 ℃ for 2h to obtain a frozen product, adjusting the temperature of the plate layer to-25 ℃, setting the vacuum degree to 0.1mbar, keeping for 4h, heating the plate layer to-15 ℃ for 3h, and heating the plate layer to 0 ℃ for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 4h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Comparative example 1
The collagen liposome lyophilized milk of this comparative example was prepared by the following steps:
According to the parts by weight, 10 parts of collagen liposome, 2 parts of mannitol, 0.5 part of beta-cyclodextrin and 87.5 parts of purified water are respectively weighed. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer to be 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, obtaining a frozen product after the freezing temperature is-40 ℃ and the freezing time is 2.5h, adjusting the temperature of the plate layer to be-22.5 ℃, setting the vacuum degree to be 0.3mbar, keeping the temperature for 4h, heating the plate layer to be-15 ℃, keeping the temperature for 3h, and heating the plate layer to be 0 ℃ and keeping the temperature for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 3h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Comparative example 2
The collagen liposome lyophilized milk of this comparative example was prepared by the following steps:
10 parts of collagen liposome, 7.5 parts of mannitol, 1.25 parts of beta-cyclodextrin and 81.25 parts of purified water are respectively weighed according to parts by mass. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution with mass fraction of 10%, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer at 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, wherein the freezing temperature is-40 ℃, the freezing time is 1h to obtain a frozen product, adjusting the temperature of the plate layer to-22.5 ℃, setting the vacuum degree to 0.3mbar, keeping for 4h, heating the plate layer to-15 ℃, keeping for 3h, and heating the plate layer to 0 ℃ for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 3h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Comparative example 3
The collagen liposome lyophilized milk of this comparative example was prepared by the following steps:
10 parts of collagen liposome, 7.5 parts of mannitol, 1.25 parts of beta-cyclodextrin and 81.25 parts of purified water are respectively weighed according to parts by mass. Adding collagen liposome into purified water, stirring and dissolving at normal temperature to obtain collagen liposome solution with mass fraction of 10%, adding mannitol into collagen liposome solution, stirring and dissolving at normal temperature to obtain mixed solution A, adding beta-cyclodextrin into mixed solution A, stirring and dissolving at normal temperature to obtain final mixed solution. And (3) placing the final mixed solution in a freeze dryer, setting the temperature of the plate layer at 5 ℃, pre-cooling for 30min, then starting a pre-freezing program, wherein the freezing temperature is-40 ℃, the freezing time is 3h to obtain a frozen product, adjusting the temperature of the plate layer to-22.5 ℃, setting the vacuum degree to 0.8mbar, keeping for 4h, heating the plate layer to-15 ℃, keeping for 3h, and heating the plate layer to 0 ℃ and keeping for 2h. And automatically entering a secondary drying stage after the primary drying is finished, setting the temperature of the plate layer to be raised to 15 ℃, keeping the temperature for 1h, setting the vacuum degree to be 0.01mbar, heating the plate layer to 30 ℃, keeping the temperature for 3h, and taking out the plate layer to obtain the collagen liposome freeze-dried emulsion.
Table 1 performance data of collagen liposome lyophilized milk prepared in each example and comparative example
Note that: the redissolution capacity index in the table: ++ means that the mixture can be redissolved by slight shaking; + represents that the solution can be redissolved only by shaking with force; -indicating that the forced shaking is also redissolved but at a slower rate; meaning that they do not dissolve after vigorous shaking.
As can be seen from Table 1, the collagen liposome lyophilized emulsion prepared in examples 1-3 can form a loose and porous dried product by adopting a lyophilization process and matching with mannitol skeleton agent, thereby ensuring the stability of the product and solving the problems of instability of the liposome in an aqueous environment, leakage in solution, phospholipid hydrolysis and the like. The beta-cyclodextrin surrounds the bioactive material in the drying process, so that the stability of the bioactive material structure is protected, the strength of a loose porous structure is improved, and the degradation of collagen is effectively prevented.
It will be apparent to those skilled in the art from this disclosure that various other changes and modifications can be made which are within the scope of the invention as defined in the appended claims.
Claims (2)
1. The collagen liposome freeze-dried emulsion is characterized by comprising the following components in parts by weight based on 100 parts by weight: 10 parts of collagen liposome, 5-10 parts of mannitol, 0.5-2 parts of beta-cyclodextrin and 78-84.5 parts of purified water, and the collagen liposome is prepared by the following steps:
(1) Adding collagen liposome into purified water according to a certain proportion, stirring and dissolving at normal temperature to obtain collagen liposome solution, then sequentially adding mannitol and beta-cyclodextrin, stirring and dissolving at normal temperature to obtain mixed solution, wherein the collagen is recombinant human collagen and is produced by fermenting Pichia pastoris with a preservation number of CGMCC No. 5021;
(2) Placing the mixed solution in a freeze dryer, setting a cooling temperature of 5 ℃, pre-cooling for 30min, and then starting a pre-freezing procedure, wherein the freezing temperature is-40 ℃, and the freezing time is 2-3 h;
(3) After the pre-freezing beam, the procedure automatically enters a primary drying stage, the vacuum degree is set to be 0.1-0.5 mbar, the plate layer temperature is set to be minus 25-minus 20 ℃, the holding time is set to be 4 hours, the plate layer temperature is increased to minus 15 ℃ and the holding time is set to be 3 hours, the plate layer temperature is increased to be 0 ℃ and the holding time is set to be 2 hours;
(4) And after the primary drying is finished, the procedure automatically enters a secondary drying stage, the temperature of the plate layer is set to be raised to 15 ℃, the holding time is 1h, the vacuum degree is set to be 0.01mbar, the temperature of the plate layer is raised to 30 ℃, and the holding time is 4h, so that the collagen liposome freeze-dried milk is obtained.
2. The collagen liposome lyophilized milk according to claim 1, wherein the average particle size of collagen liposome vesicles is 300-320 nm.
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| CN102443057A (en) * | 2011-10-26 | 2012-05-09 | 南京理工大学 | Recombinant humanized collagen and its preparation method |
| CN103110935A (en) * | 2013-01-25 | 2013-05-22 | 江苏圣宝罗药业有限公司 | Hydrolyzed collagen liposome for local skin and preparation method thereof |
| CN103536452A (en) * | 2013-11-06 | 2014-01-29 | 四川大学 | Active collagen-beta-cyclodextrin facial mask and preparation method thereof |
| CN108785128A (en) * | 2018-07-04 | 2018-11-13 | 肇庆市创业帮信息技术有限公司 | A kind of Hydrolyzed Collagen liposome and preparation method thereof of polypropylene glycol modification |
| CN111854339A (en) * | 2020-07-03 | 2020-10-30 | 江苏聚源医疗技术有限公司 | Freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102443057A (en) * | 2011-10-26 | 2012-05-09 | 南京理工大学 | Recombinant humanized collagen and its preparation method |
| CN103110935A (en) * | 2013-01-25 | 2013-05-22 | 江苏圣宝罗药业有限公司 | Hydrolyzed collagen liposome for local skin and preparation method thereof |
| CN103536452A (en) * | 2013-11-06 | 2014-01-29 | 四川大学 | Active collagen-beta-cyclodextrin facial mask and preparation method thereof |
| CN108785128A (en) * | 2018-07-04 | 2018-11-13 | 肇庆市创业帮信息技术有限公司 | A kind of Hydrolyzed Collagen liposome and preparation method thereof of polypropylene glycol modification |
| CN111854339A (en) * | 2020-07-03 | 2020-10-30 | 江苏聚源医疗技术有限公司 | Freeze-drying method of collagen-containing skin-whitening, anti-wrinkle and skin-care freeze-dried powder |
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