CN103110935A - 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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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, repair after the skin injury that is mainly used in promoting a variety of causes to cause.
Background technology
Collagen protein is the key component of human body skin skin corium, and it has the effect that moisture is retained in 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.Lack collagen protein in skin, connection will occur and solidify in collagen fiber, and the 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 a series of catabiosis such as mottle, wrinkle can appear in skin.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, directly uses, and is difficult to directly be absorbed by skin.Collagen protein is through forming the hydrolytic collagen of molecular weight after being hydrolyzed, the skin that can more be conducive to collagen protein absorbs, and the molecular weight of the hydrolytic collagen of using in cosmetics requires to be generally 1000-5000 dalton (D).Test shows, the mean molecule quantity of the best result of use of performance 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 the hydrolytic collagen polypeptide chain, therefore skin is had good moisture-keeping function.
2. reparation: collagen protein has very important effect in wound healing and tissue reconstruction process, collagen protein can promote the epidermis cell normal growth, has unique automatic repair function, the guiding epithelial cell defective region of moving into, irritation cell division, its catabolite can be by the synthetic new collagens of regenerative cell utilization, are filled in the inhibition contact that collagen between regenerative cell can reduce auxocyte, promote skin and neural the growth, and rise when cell migration and support and lubrication.The active collagen and the several amino acids that contain in collagen plaster dressing simultaneously provide nutrition for Skin Cell, the metabolism that be conducive to improve the 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 the skin allergy symptom, promotes skin metabolism, can effectively block the chain reaction that the anaphylactogen free radical causes, thereby play the effect of removing free radical, the Skin Cell film is tended towards stability, strengthen the immune system of skin, reduce allergic generation.
Relevant with growth, division, differentiation, propagation and the migration of Skin Cell by the visible hydrolytic collagen of above discussion, hydrolytic collagen can provide the skin nutrient, and delay skin aging promotes the 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 the beauty and skin care effect of performance.
The prerequisite of hydrolytic collagen performance skin repair effect is that collagen protein can see through keratodermatitis and enters skin inside, hydrolytic collagen transdermal absorption factor and molecular weight are closely related, molecular weight is less, the skin that more is conducive to hydrolytic collagen absorbs, but molecular weight is too small, water solublity increases, and particularly the free amino acid ratio increases, and also is unfavorable for the skin absorption of hydrolytic collagen and the performance of effect.Therefore the application of hydrolytic collagen is except the hydrolytic collagen of selecting 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 the 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, and take cellulose derivative, wetting agent, starch and cross-linking agent as auxiliary material, the preparation collagen mask.
But the technology that existing public technology adopts is the collagen protein simply dissolving in solution or mask matrix, directly uses, so the transdermal absorption factor of collagen protein is difficult to guarantee, directly affects the result of use of product.
In order to overcome the defective of prior art, we adopt liposome technology parcel hydrolytic collagen to prepare the hydrolytic collagen liposome, increase Transdermal absorption and the skin hold-up of hydrolytic collagen.At present about liposome technology, a large amount of bibliographical informations is arranged, but generally for protein and peptide drugs, its molecular weight ratio is larger, so 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 the hydrolytic collagen liposome, and add the hyaluronic acid of high molecular in liposome solutions, prepare the hydrolytic collagen liposome of high envelop rate, high stability, this liposome stability is good, be difficult in storage process leaking, can be for reparation and the cosmetics after all kinds of skin injurys.
Summary of the invention
The object of the invention is to provide a kind of hydrolytic collagen liposome solutions that contains, and it is characterized in that its employing phospholipid, ceramide, cholesterol prepare the hydrolytic collagen liposome, and the liposome solutions envelop rate is high, and good stability is difficult in storage process leaking.
For better understanding content of the present invention, at first we be explained as follows some technical terms.
" envelop rate " refer in liposome solutions, and bag is stated from liposome the ratio of hydrolytic collagen total amount in hydrolytic collagen amount and solution.
According to the present invention, its described hydrolytic collagen liposome solutions, contain phosphorus, ceramide, cholesterol in the lipide component of composition liposome, wherein ceramide is as one of important component of horny layer, has moisturizing, keeps 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, ceramide itself has barrier action, adhesive effect, moisture-keeping function, the anti-aging effects of skin in addition, can jointly bring into play the effect that promotes that skin injury is repaired by the synergetic hydrolysis collagen protein.
According to the present invention, described liposome solutions contains the hyaluronic acid of high molecular, studies by lot of experiments, unexpected find to have added hyaluronic acid in liposome solutions after, 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 that form extracellular matrix, content is maximum in Person's skin skin corium and knuckle synovia, 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 prepares the liposome hydrolytic collagen, and finds that in experimental study hyaluronic acid can delay liposome and leak.Creativeness will make up the skin protection composition, liposome prepares composition and the liposome stable elements combines, and hydrolytic collagen solution a kind of high percutaneous permeability, high stability and that contain the 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 comprised of cholesterol, phospholipid and ceramide, contains in its every 1000mg liposome solutions
(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 using in cosmetics requires to be generally 1000-5000 dalton (D).Hydrolytic collagen molecular weight ranges 1000-2000 dalton of the present invention (D).And the package action by liposome, improved Transdermal absorption and the skin hold-up of hydrolytic collagen.
Liposome material of the present invention is comprised 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 that causes because of ultraviolet radiation, thereby help skin anti-aging.Ceramide also helps to dispel 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 the molecular weight dependency, should according to the difference of application target, select suitable molecular weight.In Person's skin skin corium 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 the discovery, after adding hyaluronic acid in hydrolytic collagen liposome solutions of the present invention, 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 helps the recovery of skin injury on the other hand.
Liposome of the present invention, the liposome material is comprised 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, contains antiseptic 1mg-30mg in every 1000mg liposome solutions.Antiseptic is selected from and is 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 the hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion.Then remove ether under the reduction vaporization condition, obtain the gel state material, then drip the 0.1MpH6.0 phosphate buffer in the above-mentioned substance, vortex shakes just that gel comes off, and continues evaporation under decompression state, namely gets the hydrolytic collagen liposome solutions.Separately get hyaluronic acid and be added to dissolve complete in liposome solutions, namely get and contain hyaluronic liposome solutions.
Liposome solutions of the present invention can directly be applied to skin, also can add pharmaceutically acceptable adjuvant, further is 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, strengthened the skin repair effect of hydrolytic collagen.Contain in addition hydrolytic collagen, ceramide, the compound skin protection composition of hyaluronic acid in liposome solutions of the present invention, the performance synergism better promotes skin repair.
Following examples are mainly be used to 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 the hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion.Then remove ether under the reduction vaporization condition, obtain the gel state material, drip the 0.1MpH6.0 phosphate buffer in above-mentioned substance, the vortex concussion comes off gel, continues evaporation under decompression state, and get final product.Separately get hyaluronic acid, antiseptic is added in solution and dissolves, and makes liposome solutions.
Measure envelop rate and the granularity of medicine, the results are shown in following table.
In the liposome solutions of gained, the 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 is unexpected finds that hyaluronic acid can obviously improve the stability of hydrolytic collagen, delays to leak.In order better to illustrate the present invention, we have carried out stable comparative study to containing hyaluronic liposome solutions and not 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 do not contain hyaluronic hydrolytic collagen liposome, and poor stability in storage process, in storage process, envelop rate reduces greatly.
Test two: animal skin is scalded and is recovered test
Set up the animal skin damage model, illustrate the hydrolytic collagen liposome solutions for the 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 was 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
The C group is hydrolytic collagen solution (hydrolytic collagen concentration: 2%W/W) group: be coated with hydrolytic collagen solution
The D group is hyaluronic acid solution (hyaluronic acid concentration: 0.1%W/W) group: be coated with hyaluronic acid solution
Investigate index: { the minimizing percentage rate of scalded skin area=[skin lesion area before (the skin lesion area after the skin lesion area-medication before medication)/medication] * 100%} estimates 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 the B group is compared with A group, C group, D group, and significant difference (P＜0.05) is arranged, and the curative effect that the B group be described is better than A group, C group, D and organizes; After administration 14 days, B group, C group, D group are compared with the A group, and significant difference (P＜0.05) is arranged, and illustrate that the curative effect of B group, C group, D group is better than negative control group; The B group is compared with C group, D group, and significant difference (P＜0.05) is arranged, and 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 are compared with the A group, and significant difference (P＜0.05) is arranged, and illustrate that the curative effect of B group, C group, D group is better than negative control group; The B group is compared with C group, D group, and significant difference (P＜0.05) is arranged, and 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 help skin injury to recover, and hydrolytic collagen liposome solutions curative effect is obviously owing to separating collagen protein solution, hyaluronic acid solution.
Claims (10)
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 comprised of cholesterol, phospholipid and ceramide, contains in its every 1000mg liposome solutions:
(1) hydrolytic collagen 10-50mg;
(2) phosphatidase 15 0mg-200mg;
(3) ceramide 3 0-100mg;
(4) cholesterol 40-100mg;
(5) hyaluronic acid 0.5mg-5mg;
Wherein said phospholipid comprises one or more in HSPC, distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, dimyristoyl phosphatidyl choline, DLPC.
2. according to claim 1ly it is characterized in that described hydrolytic collagen molecular weight ranges 1000-2000 dalton (D) for local skin hydrolytic collagen proteolipid liquid solution.
3. according to claim 1 for local skin hydrolytic collagen proteolipid liquid solution, its described phospholipid is HSPC.
4. according to claim 1ly it is characterized in that described hyaluronan molecule weight range 100-200 ten thousand dalton (D) for local skin hydrolytic collagen proteolipid liquid solution.
5. according to claim 1ly it is characterized in that for local skin hydrolytic collagen proteolipid liquid solution, the part by weight of phospholipid and ceramide is 1: 1-4: 1; The ratio of phospholipid and cholesterol is 1: 1-3: 1.
6. want 1 describedly to it is characterized in that for local skin hydrolytic collagen proteolipid liquid solution according to right, all more than 80%, mean diameter is less than 500nm for the envelop rate of its described liposome solutions.
7. want 1 describedly to it is characterized in that for local skin hydrolytic collagen proteolipid liquid solution according to right, its described liposome solutions contains antiseptic, contains antiseptic 1mg-30mg in every 1000mg liposome solutions.
8. according to claim 7ly it is characterized in that for local skin hydrolytic collagen proteolipid liquid solution, its described antiseptic is benzalkonium bromide, Nipagin ester, benzyl alcohol, sodium benzoate.
9. according to claim 1ly it is characterized in that for local skin hydrolytic collagen proteolipid liquid solution, its described liposome solutions 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.
10. the method for preparing hydrolytic collagen liposome solutions claimed in claim 9, it is characterized in that following these steps to carrying out: phospholipid, ceramide and cholesterol are dissolved in ether, separately get hydrolytic collagen, be dissolved in water, the diethyl ether solution that is dissolved with phospholipid, ceramide and cholesterol is added in the hydrolytic collagen aqueous solution, through ultrasonic or concussion in short-term, form stable w/o type Emulsion.Then remove ether under the reduction vaporization condition, obtain the gel state material, then drip the 0.1MpH6.0 phosphate buffer in above-mentioned substance, the vortex concussion comes off gel, continue evaporation under decompression state, separately get hyaluronic acid and be added in solution and dissolve, make liposome solutions.
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| CN106390093A (en) * | 2015-07-28 | 2017-02-15 | Zo皮肤健康公司 | Post-procedure skin care systems, compositions, and methods of use thereof |
| CN107441046A (en) * | 2017-08-31 | 2017-12-08 | 宁波盖济尔医疗科技有限公司 | A kind of compound protein liposome nanometer formulation |
| 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 |
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| CN106390093B (en) * | 2015-07-28 | 2019-04-05 | Zo皮肤健康公司 | Skin nursing system, composition and its application method after operation |
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| 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 |
| CN115177535A (en) * | 2022-05-27 | 2022-10-14 | 吉林省中鹿中医药产业(集团)有限公司 | Deer skin collagen polypeptide liposome hydrogel, preparation method and application 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 | Deer skin collagen polypeptide liposome hydrogel, preparation method and application thereof |
| CN116270296A (en) * | 2023-04-13 | 2023-06-23 | 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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