CN103110935B - Hydrolyzed collagen liposome for local skin and preparation method thereof - Google Patents
Hydrolyzed collagen liposome for local skin and preparation method thereof Download PDFInfo
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- CN103110935B CN103110935B CN201310030618.8A CN201310030618A CN103110935B CN 103110935 B CN103110935 B CN 103110935B CN 201310030618 A CN201310030618 A CN 201310030618A CN 103110935 B CN103110935 B CN 103110935B
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Abstract
The invention provides a hydrolyzed collagen liposome solution mainly used for repairing skin damages caused by various factors. According to the invention, the hydrolyzed collagen liposome solution is prepared from a phospholipid, a ceramide and cholesterol, a percutaneous stratum corneum of hydrolyzed collagen is promoted to enter the skin, and the retention of the hydrolyzed collagen in the skin is improved, so that the recovery of the skin damages is effectively promoted. Furthermore, the solution contains hyaluronic acid to improve the stability of the liposome and delay leakage.
Description
Technical field
The present invention relates to a kind of local skin hydrolytic collagen proteoliposome, after the skin injury that is mainly used in promoting a variety of causes to cause, repair.
Background technology
Collagen protein is the key component of human body skin skin corium, and it has the effect that moisture is retained in to skin corium, and provides support and protect, and gives skin elasticity and intensity.With advancing age, the collagen protein in skin slowly metabolism reduces, and ultraviolet reaches skin corium by epidermis in addition, impels collagen protein to decompose.In skin, lack collagen protein, will there is connection and solidify in collagen fiber, iuntercellular mucopolysaccharide is reduced, and skin just can lose softness, elasticity and gloss, the fibrous fracture of corium, lipoatrophy, sweat gland and smegma reduce simultaneously, and skin there will be a series of catabiosis such as mottle, wrinkle.The relative molecular mass of collagen protein is 300,000 dalton (D), and it is the triple-helix structure that is combined into and is interwinded by 3 peptide chains, and directly application is difficult to directly be absorbed by skin.Collagen protein forms the hydrolytic collagen of molecular weight after hydrolysis, and the skin that can more be conducive to collagen protein absorbs, and the molecular weight of the hydrolytic collagen of applying in cosmetics requires to be generally 1000-5000 dalton (D).Test shows, the mean molecule quantity of bringing into play best result of use should be in 700-1000 dalton (D) scope.
Hydrolytic collagen can be applicable in all kinds of skin repair medical supplies and cosmetics, and its major physiological activity shows as:
1. moisture retention: owing to containing the hydrophilic groups such as amino, carboxyl and hydroxyl in hydrolytic collagen polypeptide chain, therefore skin is had to good moisture-keeping function.
2. reparation property: collagen protein has very important effect in wound healing and tissue reconstruction process, collagen protein can promote epidermis cell normal growth, there is unique automatic repair function, the guiding epithelial cell defective region of moving into, irritation cell division, its catabolite can be by the synthetic new collagen of regenerative cell utilization, is filled in collagen between regenerative cell and can reduces the inhibition contact of auxocyte, promote skin and neural growth, and in the time of cell migration, rise and support and lubrication.The active collagen simultaneously containing in collagen plaster dressing and several amino acids provide nutrition for Skin Cell, the metabolism that be conducive to improve epidermis cell microcirculation, promotes skin, thereby accelerate tissue regeneration and recovery, promote the epithelial proliferation for repairing of wound surface, the damage of repairing hair follicle wall, alleviates follicular wall rupture.
3. the speckle effect of discoloring: collagen protein has the skin of making and keeps moistening, alleviates skin allergy symptom, promotes skin metabolism, can effectively block the chain reaction that anaphylactogen free radical causes, thereby play the effect of removing free radical, Skin Cell film is tended towards stability, the immune system that strengthens skin, reduces allergic generation.
By discussing above, growth, division, differentiation, propagation and the migration of visible hydrolytic collagen with Skin Cell be relevant, and hydrolytic collagen can provide skin nutrient, and delay skin aging promotes skin wound reparation.Experimentation and clinical practice all show, hydrolytic collagen is added in skin repair articles for use, cosmetics, can effectively have an effect with Skin Cell, promote the reparation of injured skin and beauty and skin care effect of performance.
The prerequisite of hydrolytic collagen performance skin repair effect is that collagen protein can enter skin inside through keratodermatitis, hydrolytic collagen transdermal absorption factor and molecular weight are closely related, molecular weight is less, the skin that is more conducive to hydrolytic collagen absorbs, but molecular weight is too small, water solublity increases, and particularly free amino acid ratio increases, and is also unfavorable for the skin absorption of hydrolytic collagen and the performance of effect.Therefore the application of hydrolytic collagen is except selecting the hydrolytic collagen of suitable molecular weight, and adopting transdermal to urge to ooze the absorption of technology facilitation of hydrolysis collagen protein is also the emphasis direction of research.
Chinese patent CN1219505C discloses a kind of collagen protein face paper and preparation method thereof, and preparation technology is simple, can be used for facial nursing needs.
Chinese patent CN100460516C discloses a kind of preparation method of collagen protein face, is primarily characterized in that take collagen protein as functional raw material, take cellulose derivative, wetting agent, starch and cross-linking agent as auxiliary material, prepares collagen mask.
But the technology that existing public technology adopts is collagen protein simply dissolving in solution or mask matrix, directly uses, therefore the transdermal absorption factor of collagen protein is difficult to guarantee, directly affects the result of use of product.
In order to overcome the defect of prior art, we adopt liposome technology parcel hydrolytic collagen to prepare hydrolytic collagen liposome, increase Transdermal absorption and the skin hold-up of hydrolytic collagen.At present have a large amount of bibliographical informations about liposome technology, but generally for protein and peptide drugs, its molecular weight ratio is larger, therefore, in encapsulation process, envelop rate is difficult to improve, and in storage process, medicine is easy seepage also.The present invention adopts phospholipid, ceramide and cholesterol parcel hydrolytic collagen, prepare hydrolytic collagen liposome, and in liposome solutions, add the hyaluronic acid of high molecular, prepare the hydrolytic collagen liposome of high envelop rate, high stability, this liposome stability is good, in storage process, be difficult for leaking, can be for the reparation after all kinds of skin injurys and cosmetics.
Summary of the invention
The object of the invention is to provide one and contains hydrolytic collagen liposome solutions, it is characterized in that it adopts phospholipid, ceramide, cholesterol to prepare hydrolytic collagen liposome, and liposome solutions envelop rate is high, and good stability is difficult in storage process leaking.
For better understanding content of the present invention, first we be explained as follows some technical terms.
" envelop rate " refer in liposome solutions, and bag is loaded in liposome the ratio of hydrolytic collagen total amount in hydrolytic collagen amount and solution.
According to the present invention, hydrolytic collagen liposome solutions described in it, in the lipide component of composition liposome, contain phosphorus, ceramide, cholesterol, wherein ceramide is as one of important component of horny layer, has moisturizing, maintains skin barrier, the physiological function such as defying age, antiallergic, cell death inducing.Therefore liposome of the present invention not only can improve Transdermal absorption and the skin hold-up of hydrolytic collagen, in addition ceramide itself has barrier action, adhesive effect, moisture-keeping function, the anti-aging effects of skin, can jointly bring into play the effect that promotes that skin injury is repaired by synergetic hydrolysis collagen protein.
According to the present invention, described liposome solutions, the hyaluronic acid that contains high molecular, studies by lot of experiments, and unexpected discovery added after hyaluronic acid in liposome solutions, greatly increased the stability of liposome, reduced drug leakage.Hyaluronic acid is the straight chain polymer polysaccharide that disaccharidase unit forms by glucuronic acid-NAG, be present in widely in the connective tissue of organism, one of several glycosaminoglycans of composition extracellular matrix, in people's dermal layer of the skin and knuckle synovia, content is maximum, has water conservation, the important physiological action such as lubricated.
According to the present invention, described liposome solutions contains the skin-nourishing compositions such as hydrolytic collagen, ceramide, hyaluronic acid simultaneously.The lipid that utilization contains ceramide is prepared liposome hydrolytic collagen, and in experimental study, finds that hyaluronic acid can delay liposome and leak.Combination skin protection composition, liposome are prepared to composition to creativeness and liposome stable elements combines, and hydrolytic collagen solution a kind of high percutaneous permeability, high stability and that contain skin protection combination ingredient is provided.
Hydrolytic collagen liposome solutions of the present invention, described liposome solutions comprises hydrolytic collagen liposome and hyaluronic acid, described hydrolytic collagen liposome comprises hydrolytic collagen and filmogen, described filmogen is made up of cholesterol, phospholipid and ceramide, in its every 1000mg liposome solutions, contains
(1) hydrolytic collagen 10-50mg;
(2) phosphatidase 15 0mg-200mg;
(3) ceramide 3 0-100mg;
(4) cholesterol 40-150mg;
(5) hyaluronic acid 0.5mg-5mg;
Wherein said phospholipid comprises one or more in HSPC, distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, dimyristoyl phosphatidyl choline, DLPC.
The molecular weight of hydrolytic collagen is relevant to its transdermal ability, and the hydrolytic collagen molecular weight of applying in cosmetics requires to be generally 1000-5000 dalton (D).Hydrolytic collagen molecular weight ranges 1000-2000 dalton of the present invention (D).And by the package action of liposome, improve Transdermal absorption and the skin hold-up of hydrolytic collagen.
Liposome material of the present invention is made up of lipid and cholesterol, and wherein lipid comprises phospholipid and ceramide.Ceramide is a kind of lipid components in normal biofilm structure, is distributed in the skin of Lipid Bilayer Structure.Ceramide can strengthen the cohesiveness of epidermis cell, repair skin barrier function, thereby alleviate the desquamation symptom of angleplied laminate, help skin to recover again, improve skin appearance, make skin smooth flexible, also can avoid or reduce the abrasion causing because of ultraviolet radiation, thereby contribute to skin anti-aging.Ceramide also contributes to dispel the wrinkle on skin, removes skin dermatitis, keeps the elasticity of skin.
Hydrolytic collagen liposome solutions of the present invention contains hyaluronic acid.Hyaluronic acid is a kind of macromole mucopolysaccharide, and its biological activity and result of use have molecular weight dependency, should, according to the difference of application target, select suitable molecular weight.In people's dermal layer of the skin and knuckle synovia, hyaluronic acid contents is maximum, has water conservation, the important physiological action such as lubricated.Due to the character of its unique water conservation, the macromolecule polysaccharide such as lubricated, its natural sex, without anaphylaxis and the biocompatibility good with tissue; Molecular weight has good moisture retention, lubricated and medicament slow release effect at 1,000,000-200 dalton's (D) hyaluronic acid, can be widely used in cosmetics, eye drop, skin burn concrescence and post-operation adhesion preventing.By testing unexpected discovery, in hydrolytic collagen liposome solutions of the present invention, add after hyaluronic acid, can greatly improve the stability of liposome, delay to leak.
Hyaluronan molecule weight range 100-200 of the present invention ten thousand dalton (D), hyaluronic acid can increase the stability of liposome on the one hand; Hyaluronic acid contributes to the recovery of skin injury on the other hand.
Liposome of the present invention, liposome material is made up of phospholipid, ceramide, cholesterol, and wherein the part by weight of phospholipid and ceramide is 1: 1-4: 1; The ratio of phospholipid and cholesterol is 1: 1-3: 1.
Hydrolytic collagen liposome solutions envelop rate of the present invention is greater than 80%, and mean diameter is less than 500nm.
Liposome solutions of the present invention contains antiseptic, in every 1000mg liposome solutions, contains antiseptic 1mg-30mg.Antiseptic is selected from as benzalkonium bromide, Nipagin ester, benzyl alcohol, sodium benzoate.
Liposome solutions of the present invention, is characterized in that having following composition:
(1) hydrolytic collagen 20mg;
(2) HSPC 70mg;
(3) DLPC 30mg;
(4) ceramide 50mg;
(5) cholesterol 50mg;
(6) hyaluronic acid 1mg;
(7) antiseptic 1mg-30mg;
(8) 0.1M pH6.0 phosphate buffer adds to 1000mg.
Liposome solutions of the present invention, follow these steps to carry out: phospholipid, ceramide, cholesterol are dissolved in ether, separately get hydrolytic collagen, be dissolved in water, the diethyl ether solution that is dissolved with phospholipid is added in hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion.Then under reduction vaporization condition, remove ether, obtain gel state material, then drip 0.1MpH6.0 phosphate buffer in above-mentioned substance, vortex shakes just gel and comes off, and continues evaporation under decompression state, obtains hydrolytic collagen liposome solutions.Separately get hyaluronic acid and be added in liposome solutions and dissolve completely, must contain hyaluronic liposome solutions.
Liposome solutions of the present invention, can directly be applied to skin, also can add pharmaceutically acceptable adjuvant, is further prepared into ointment, gel, is convenient to clinical using.
The advantage of the present invention and existing technology is: adopt phospholipid, ceramide, cholesterol to prepare the hydrolytic collagen liposome of high envelop rate, high stability as matrix material, liposome encapsulation is greater than 80%, and mean diameter is less than 500nm.By the package action of liposome, improve transdermal absorption factor and the skin hold-up of hydrolytic collagen, strengthen the skin repair effect of hydrolytic collagen.In liposome solutions of the present invention, contain in addition hydrolytic collagen, ceramide, the compound skin protection composition of hyaluronic acid, performance synergism, better promotes skin repair.
Following examples are mainly used for further illustrating the present invention, rather than limit the scope of the invention.
Embodiment 1: the preparation of hydrolytic collagen liposome solutions
The prescription table of table 1 hydrolytic collagen liposome solutions
Preparation technology: phospholipid, ceramide, cholesterol are dissolved in ether, separately get hydrolytic collagen, be dissolved in water, the diethyl ether solution that is dissolved with phospholipid is added in hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion.Then under reduction vaporization condition, remove ether, obtain gel state material, in above-mentioned substance, drip 0.1MpH6.0 phosphate buffer, vortex concussion comes off gel, continues evaporation under decompression state, to obtain final product.Separately get hyaluronic acid, antiseptic is added in solution and dissolves, and makes liposome solutions.
Envelop rate and the granularity of measuring medicine, the results are shown in following table.
In the liposome solutions of gained, hydrolytic collagen envelop rate is greater than 80%; Particle mean size is less than 500nm.
Test one: the investigation of liposome solutions stability
Hydrolytic collagen is wrapped in liposome, and liposome can carry hydrolytic collagen and penetrate into skin, the effect of performance skin repair.The present invention surprisingly finds that hyaluronic acid can obviously improve the stability of hydrolytic collagen, delays to leak.In order better to illustrate the present invention, we are to containing hyaluronic liposome solutions and not carried out stability comparative study containing hyaluronic liposome solutions.
Press the formulation and technology of embodiment 2, preparation contains hyaluronic hydrolytic collagen liposome solutions and does not contain hyaluronic hydrolytic collagen liposome solutions respectively.Get 2 kinds of hydrolytic collagen liposome solutions and be placed in 30 ℃ of placements, respectively at 0 month, January, February, March, sampling in June, measure and the hydrolytic collagen envelop rate of comparison solution.
Shown by upper watch test result, contain hyaluronic hydrolytic collagen liposome, good stability, does not leak in storage process.And not containing hyaluronic hydrolytic collagen liposome, poor stability in storage process, in storage process, envelop rate reduces greatly.
Test two: animal skin is scalded and recovered test
Set up animal skin damage model, illustrate hydrolytic collagen liposome solutions for skin injury repair.Research as a comparison, we have prepared common hydrolytic collagen protein solution and the hyaluronic acid solution sample of same concentrations.Relatively liposome solutions, common hydrolytic collagen protein solution and hyaluronic acid solution are repaired the difference of damaged skin curative effect.
Animal Model: after rat back is lost hair or feathers, next day is under pentobarbital sodium 30mg/kg anesthesia, adopt under 80 ℃ of conditions of water bath with thermostatic control scald apparatus, continue 6 seconds, cause shallow II degree to scald plucked skin of back, area is 5cm × 8cm, is approximately 10% of body surface area.
Medication: for the ease of coating and administration, use sponge absorption given the test agent, spread on damaged skin, drug of topical application every day 1 time, each 2 hours, successive administration 14 days, observed injured skin area recovery situation.
Experiment grouping: be divided into 4 groups, 12 every group.
A organizes negative control group: be coated with normal saline
B group hydrolytic collagen liposome group (hydrolytic collagen concentration: 2%W/W; Hyaluronic acid concentration: 0.1%W/W): be coated with embodiment 2 hydrolytic collagen liposome solutions
C group is hydrolytic collagen solution (hydrolytic collagen concentration: 2%W/W) group: be coated with hydrolytic collagen solution
D group is hyaluronic acid solution (hyaluronic acid concentration: 0.1%W/W) group: be coated with hyaluronic acid solution
Investigate index: { minimizing percentage rate=[skin lesion area before (the skin lesion area after the skin lesion area-medication before medication)/medication] × 100%} of scalded skin area, evaluates drug effect with this to the minimizing percentage rate of calculating scalded skin area.
The results are shown in following table:
Result shows, administration is after 7 days, and B group, compared with A group, C group, D group, has significant difference (P < 0.05), illustrates that the curative effect of B group is better than A group, C group, D group; After administration 14 days, B group, C group, D group, compared with A group, have significant difference (P < 0.05), illustrate that the curative effect of B group, C group, D group is better than negative control group; B group, compared with C group, D group, has significant difference (P < 0.05), illustrates that the curative effect of B group is better than C group, D group; After administration 21 days, B group, C group, D group, compared with A group, have significant difference (P < 0.05), illustrate that the curative effect of B group, C group, D group is better than negative control group; B group, compared with C group, D group, has significant difference (P < 0.05), illustrates that the curative effect of B group is better than C group, D group.
Conclusion: hydrolytic collagen liposome solutions, hydrolytic collagen solution, hyaluronic acid solution contribute to skin injury to recover, hydrolytic collagen liposome solutions curative effect is obviously owing to separating collagen solution, hyaluronic acid solution.
Claims (2)
1. one kind for local skin hydrolytic collagen proteolipid liquid solution, it is characterized in that, described liposome solutions comprises hydrolytic collagen liposome and hyaluronic acid, described hydrolytic collagen liposome comprises hydrolytic collagen and filmogen, described filmogen is made up of cholesterol, phospholipid and ceramide, in its every 1000mg liposome solutions, contains:
2. the method for preparation hydrolytic collagen liposome solutions claimed in claim 1, it is characterized in that following these steps to carrying out: by phospholipid, ceramide and cholesterol are dissolved in ether, separately get hydrolytic collagen, be dissolved in water, to be dissolved with phospholipid, the diethyl ether solution of ceramide and cholesterol is added in hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion, then under reduction vaporization condition, remove ether, obtain gel state material, in above-mentioned substance, drip 0.1MpH6.0 phosphate buffer again, vortex concussion comes off gel, under decompression state, continue evaporation, separately get hyaluronic acid, antiseptic is added in solution and dissolves, make liposome solutions.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US9248160B1 (en) * | 2015-07-28 | 2016-02-02 | Zo Skin Health, Inc. | Post-procedure skin care systems, compositions, and methods of use thereof |
| CN107441046B (en) * | 2017-08-31 | 2020-07-28 | 宁波盖济尔医疗科技有限公司 | Composite protein liposome nano preparation |
| 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 |
| CN109394567A (en) * | 2018-10-19 | 2019-03-01 | 天津工业大学 | A kind of preparation method and quality testing standard of multifunctional transparent matter acid liposome |
| CN109394563A (en) * | 2018-11-06 | 2019-03-01 | 福建省海西细胞生物工程有限公司 | A kind of preparation method of stem cell media liposome and the application in skin care item |
| CN112315821B (en) * | 2020-12-03 | 2022-09-23 | 暨南大学 | Hydrolyzed protein liposome and preparation method and application thereof |
| CN113679631A (en) * | 2021-09-28 | 2021-11-23 | 常州金远药业制造有限公司 | Tranexamic acid liposome cosmetic containing hyaluronic acid and preparation method thereof |
| CN114177278A (en) * | 2021-10-18 | 2022-03-15 | 山东多美康生物医药有限公司 | Liposome preparation |
| CN114470057B (en) * | 2021-10-26 | 2023-12-22 | 成都科建生物医药有限公司 | Liposome skin repair preparation and preparation method thereof |
| CN114470171B (en) * | 2021-12-31 | 2024-06-11 | 江苏聚源医疗技术有限公司 | Collagen liposome freeze-dried emulsion and preparation method thereof |
| CN115177535B (en) * | 2022-05-27 | 2023-07-07 | 吉林省中鹿中医药产业(集团)有限公司 | Buckskin collagen polypeptide liposome hydrogel, preparation method and application thereof |
| CN116270296B (en) * | 2023-04-13 | 2024-10-22 | 时垠(上海)生物科技有限公司 | Collagen lipid inclusion and preparation method and application thereof |
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| JP2004323401A (en) * | 2003-04-23 | 2004-11-18 | Mikimoto Pharmaceut Co Ltd | Skin care preparation for external use |
| JP2005060234A (en) * | 2003-08-08 | 2005-03-10 | Rohto Pharmaceut Co Ltd | External preparation for skin |
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| CN102397168A (en) * | 2011-11-23 | 2012-04-04 | 苏州瑞纳生化技术有限公司 | Flexible nanoliposomes with charges for cosmetics and preparation method thereof |
| CN102429913A (en) * | 2011-12-13 | 2012-05-02 | 江苏圣宝罗药业有限公司 | Compound clobetasol propionate liposome and preparation thereof |
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