CN108721604A - A kind of -4 alcohol plastid of extrasin beta and its preparation process - Google Patents
A kind of -4 alcohol plastid of extrasin beta and its preparation process Download PDFInfo
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Abstract
The invention discloses a kind of -4 alcohol plastid of extrasin beta and preparation method thereof, -4 alcohol plastid concrete composition of the extrasin beta includes：Extrasin beta -4 0.022%~0.043%, phosphatidase 0 .054%~0.084%, cholesterol 0.018%~0.028%, anionic surfactant 0.02%~0.04%, ethyl alcohol 10.10%~30.30%, remaining is distilled water；With the Transdermal absorption for increasing drug, enhance the technique effect of the reparation of -4 pairs of scarred skins of extrasin beta.And in the present invention -4 alcohol plastid of extrasin beta preparation method, by be added anionic surfactant and cholesterol in the way of solve the stability problem of -4 alcohol plastid of extrasin beta, -4 alcohol plastid encapsulation rate of extrasin beta obtained using the preparation method is high, it has good stability, it is simple for process, it is with good economic efficiency.Correspondingly, the present invention also protects -4 alcohol plastid of above-mentioned extrasin beta in the application in scar reparation field.
Description
Technical field
The present invention relates to field of pharmaceutical preparations, and in particular to a kind of -4 alcohol plastid of extrasin beta and its preparation process and application.
Background technology
Skin injury caused by the reasons such as various wounds, burn, operation, radiation is common clinical, height morbidity.According to system
The expense of meter, global trauma care reached 11,500,000,000 dollars in 2016.With current treatment means, formed after wound healing
Rate is up to 62% (white people) and 80% (non-white man), these scars often bring long-term physiological barrier and psychology negative to patient
Load.Development can speed up wound healing, promotes the accessory organs such as wound part hair follicle, sweat gland newborn, wound tissue is helped to tie
Structure and the pharmaceutical preparation for functionally obtaining dual reparation have very great research significance.Currently, to outside wound locally supplement
The cell factor of source property is proved to play an important role in the wound repair of various organization organ.
Extrasin beta -4 is a kind of cell factor with multinomial biological function, is made of 43 amino acid multi-functional
Polypeptide is distributed widely in karyocyte, rich in in blood platelet, function combined with G-actin actins it is closely related,
It being capable of modulate actin polymerization, depolymerization.Therefore, extrasin beta -4 there is extrasin beta -4 potential clinic to answer plastic surgery
With value, has and promote the critical functions such as cell differentiation maturation, tissue injury reparation, revascularization and hair follicle regeneration, in wound
It repairs, great drug development potentiality in terms of corneal restoration and myocardial repair, but extrasin beta -4 is directly as scar repair medicine
Correlative study is carried out using also few people.
Alcohol plastid has double points of lipid similar with corneocyte by what phosphatide, cholesterol, second alcohol and water were formed
Sublayer structure, excellent biocompatibility can by it is many itself can not transdermal drug be wrapped in wherein, be carried along into skin,
It forms drug depot and persistently plays local drug effect.Alcohol plastid have bilayer mobility it is higher, it is easily deformable, conducive to penetrating
Skin barrier increases the advantages of drug accumulation in skin, compared to liposome have better Transdermal absorption performance and compared with
High encapsulation rate, one side alcohol plastid contain the skin penetration enhancer ethyl alcohol of high concentration, change Stratum corneum lipids molecule
Close-packed arrays enhance lipid fluidity；The ethyl alcohol of another aspect high concentration enhances the flexibility and mobility of alcohol plastid film, makes
Containing alcohol entrapment efficiency of liposome higher, can deform in transmittance process, overcome the barrier action of skin, through disorder
Cuticula enhances penetrating power.Alcohol plastid is applied to percutaneous dosing, the permeation rate of drugs can be significantly increased, increases skin
The cumulant of drug in skin, and do not have the correlative study of -4 alcohol plastid preparation process of extrasin beta also in the prior art.
Invention content
For the above-mentioned prior art, the present invention provides following technical scheme：
It is an object of the present invention to provide a kind of -4 alcohol plastids of extrasin beta, and extrasin beta -4 is combined with alcohol plastid, is carried
Supplied a kind of novel percutaneous preparation, said preparation have stability is good, preparation process is simple, Small side effects, transdermal effect more
Strong feature can enhance -4 cell factor transdermal effect of extrasin beta, promote the repair to scar.
The second purpose of the present invention is to provide the preparation method of -4 alcohol plastid of above-mentioned extrasin beta, the preparation side in the present invention
Method solves the stability problem of alcohol plastid by the way that anionic surfactant is added, and manufacturing cost is cheap, simple for process, surely
It is qualitative good.
The three of the object of the invention are that -4 alcohol plastid of above-mentioned extrasin beta is protected to prepare answering in terms of scar maintenance preparation
With.
To achieve the goals above, the present invention adopts the following technical scheme that：
The present invention provides a kind of -4 alcohol plastid of extrasin beta, is made of the raw material of following parts by weight：Extrasin beta-
40.022%~0.043%, phosphatidase 0 .054%~0.084%, cholesterol 0.018%~0.028%, anionic surface is lived
Property agent 0.02%~0.04%, ethyl alcohol 10.10%~30.30%, remaining is distilled water.Anionic surfactant is in we
Can not only play the role of dispersion in case, serve as the deformability that film softening agent makes it have height, it is often more important that, it is cloudy from
The addition of subtype surfactant contributes to the raising of alcohol plastid stability, significantly increases the extrasin beta-prepared in the present invention
The long-time stability of 4 alcohol plastids.
Preferably, the raw material group of -4 alcohol plastid of above-mentioned extrasin beta becomes -4 0.025-0.035% of extrasin beta, phosphatide
0.06-0.08%, cholesterol 0.02-0.025%, anionic surfactant 0.022-0.03%, ethyl alcohol 18-28%,
Remaining is distilled water.
Preferably, the raw material group of -4 alcohol plastid of above-mentioned extrasin beta becomes extrasin beta -4 0.03198%, phosphatide
0.06396%, cholesterol 0.02131%, anionic surfactant 0.02665%, ethyl alcohol 25.23%, remaining is distillation
Water.
Further, above-mentioned anion surfactant is dexycholate or cholate, it is preferred that is deoxycholic acid
Salt.
Above-mentioned phosphatide can be Fabaceous Lecithin in above-mentioned raw materials composition, can also be lecithin, it is preferred that be Fabaceous Lecithin.
The present invention also provides the preparation method of -4 alcohol plastid of above-mentioned extrasin beta, step includes：
(1) phosphatide and cholesterol are dissolved in acquisition alcohol phase solution in absolute ethyl alcohol；
(2) surfactant NaTDC and extrasin beta -4 are dissolved in distilled water and are made into aqueous phase solution；
(3) aqueous phase solution is slowly injected into aquation certain time in alcohol phase solution under stiring；
(4) miillpore filter is crossed after being cooled to room temperature the solution after aquation to get -4 alcohol plastid of extrasin beta.
Preferably, hydration temperature is 25~40 DEG C in step (3), and hydration time is 10~30min, stir speed (S.S.) 500
~800r/min, water phase charge velocity are 0.2mL/min~2mL/min.
It is found in research process of the present invention, water phase is injected in alcohol phase to be conducive to obtain and stablizes clear alcohol plastid solution,
Charge velocity, hydrating condition etc. pair and the encapsulating effect of alcohol plastid in the present invention have significant impact.
Preferably, miillpore filter aperture used is 450nm in step (4).
The present invention also provides application of -4 alcohol plastid of above-mentioned extrasin beta in terms of preparing reparation facial mask.The reparation facial mask by
Above-mentioned β -4 alcohol plastids, carbomer gel, freeze drying protectant composition.
The present invention also provides the preparation methods of above-mentioned Face-protecting mask, and carbomer gel is taken to be placed in distilled water, stir one section
Time stands overnight after the completion of stirring, so that it is fully swollen, then be slowly added into freeze drying protectant and obtain water white transparency Blank gel
Matrix.The alcohol plastid solution prepared in advance is added in Blank gel matrix, mixes uniformly both to obtain β -4 alcohol plastids maintenance face
Film.
- 4 alcohol plastid of extrasin beta has great role to scar repairing, can be applied to facial mask field, plays moisturizing and reparation
Double effects.
Preferably, carbomer gel 934 or 940 is placed in distilled water, 300~500rpm/min stirs 3~5h, stirring
After in left at room temperature over night.
Preferably, the mass ratio of card wave gel and distilled water freeze drying protectant is 3~5:17~20:0.5~2.
Preferably, above-mentioned freeze drying protectant is sucrose, glucose, one kind in mannitol or combination.
Beneficial effects of the present invention
1. the biological function of extrasin beta -4 includes regeneration, remodeling, Wound healing, maintains actin balance, is swollen
The effect that tumor is fallen ill with transfer, cells apoptosis and inflammation etc., the precedent for being directly used as scar maintenance drug are fresh
It has been reported that, a kind of -4 alcohol plastid of extrasin beta of Selecting research of the present invention and its preparation process provide one kind for scar maintenance preparation
New Research idea.
2. current alcohol plastid mainly contains, chemical drug, traditional Chinese medicine ingredients are relatively more, and fresh to contain polypeptide drug less, main cause exists
It is big in the molecular weight of polypeptide drug, and drugloading rate is too low.But -4 local administration of extrasin beta only needs lower dose to can reach
The effect of scar reparation, while alcohol plastid increases the transdermal characteristic of drug again, is that dosage form brings out the best in each other with drug, and polypeptide
Easy in inactivation after the present invention is coated extrasin beta -4 using alcohol plastid, can reduce the inactivation of polypeptide, extend active medicine
Action time.
3. the present invention has found during the experiment, cholesterol is added in preparation process and advantageously forms big single chamber lipid
Body, hydrophilic chamber are more advantageous to greatly hydrophilic drug extrasin beta -4 of package, meanwhile, cholesterol also plays the effect of " rivet ",
Keep alcohol plastid membrane structure firmer.The present invention is in the course of the research, it was found that the addition of anionic surfactant, for
The stabilization of alcohol plastid has good improved effect, by the addition of conventional reagent, achieves unexpected technique effect.
4. in the preparation process of alcohol plastid, dosage, injection mode, injection rate, drop rate of reagent etc. are to alcohol matter
The stabilization of body has important influence.For the present invention by a lot of research work, good stability alcohol matter can be prepared by providing one kind
The method of body has important dissemination.
Description of the drawings
The accompanying drawings which form a part of this application are used for providing further understanding of the present application, and the application's shows
Meaning property embodiment and its explanation do not constitute the improper restriction to the application for explaining the application.
Fig. 1 is the TEM photo figures that alcohol plastid is made in alcohol phase injection；
Fig. 2 is the TEM photo figures that alcohol plastid is made in water phase injection；
Fig. 3 is the TEM photo figures that alcohol plastid is made in different preparation methods；
Fig. 4 is the β -4 alcohol plastid grain size distributions prepared in embodiment 4；
Fig. 5 is the β -4 alcohol plastid potential images prepared in embodiment 4；
Fig. 6 is the TEM photo figures of β -4 alcohol plastids prepared in embodiment 4；
Fig. 7 is the β -4 alcohol plastid facial mask photo figures prepared in embodiment 8；
Fig. 8 is that photo figure is lyophilized in β -4 alcohol plastid facial masks in embodiment 8.
Specific implementation mode
The present invention is specifically described below by embodiment, it is necessary to be pointed out that following embodiments are served only for this
Invention is described further, and should not be understood as limiting the scope of the invention, and person skilled in art states under
Some nonessential modifications and adaptations that bright content is made are all within the scope of the present invention.
It should be noted that term used herein above is merely to describe specific implementation mode, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or combination thereof.
Transmission electron microscope photo in the present invention is JEM-2100 transmission electron microscopes (Jeol Ltd. JEOL)
Shooting.Granularmetric analysis experiment uses Zetasizer Nano ZS grain sizes potentiometric analyzers (Britain of Malvem companies).Freeze-drying
Machine is purchased from Shanghai Yu Ming Instrument Ltd (China).
As background technology is introduced, for the deficiency of scar maintenance drug in the prior art, the present invention proposes one
Kind -4 alcohol plastid of extrasin beta and preparation method.
In order to enable those skilled in the art can clearly understand the technical solution of the application, below with reference to tool
The technical solution of the application is described in detail with comparative example for the embodiment of body.
The selection of 1 preparation method of embodiment
1.1 alcohol phases are injected
During the cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg addition 1mL ethanol solutions are set, by 0.50mg extrasin betas-
4, which are dissolved in 1.75mL distilled water, is placed in pear shape bottle, alcohol phase mixed solution is injected with 400uL/min in pear shape bottle solution, 30 DEG C
Under the conditions of 700r/min aquation 15min, after being cooled to room temperature cross aperture be 450nm miillpore filter, carry out centainly handle after survey
EE is 10 ± 4%, and alcohol plastid solution appearance is muddy, and TEM photo backgrounds are dirty and messy, and molding alcohol plastid is not of uniform size, is ruptured mostly
Or be adhered, as a result as shown in Fig. 1.1.2 water phases are injected
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg addition 1mL ethanol solutions are placed in pear shape bottle, by 0.50mg
Extrasin beta -4 is dissolved in 1.75mL distilled water, water phase mixed solution is injected with 400uL/min in pear shape bottle solution, 30 DEG C of items
700r/min aquations 15min under part crosses the miillpore filter that aperture is 450nm after being cooled to room temperature, carry out surveying EE after centainly handling
It is 74 ± 2%, as shown in Fig. 2, alcohol plastid solution appearance clear has light blue opalescence to occur, TEM photo roundings.
2 alcohol plastid of embodiment prepares single factor experiment
The dosage of 2.1 cholesterol
The cholesterol of 1.5mg Fabaceous Lecithins and different amounts addition 1mL ethanol solutions are placed in pear shape bottle, it will
0.50mg extrasin betas -4 are dissolved in 1.75mL distilled water, and water phase mixed solution is injected with 400uL/min in pear shape bottle solution,
700r/min aquations 15min under the conditions of 30 DEG C crosses the miillpore filter that aperture is 450nm, is centainly handled after being cooled to room temperature
After survey EE, experiment measures that the results are shown in table below：
Influence (n=3) of the cholesterol dosage for EE
Cholesterol dosage produces strong influence for the EE of alcohol plastid, when being especially not added with cholesterol, alcohol obtained
Plastid solution cannot show light blue opalescence well, as shown in figure 3, TEM photos show only minute quantity alcohol plastid molding
And it is in irregular shape.
2.2 concentration of alcohol
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in the ethanol solution of 1mL and are placed in pear shape bottle, it will
0.50mg extrasin betas -4 are dissolved in 1.75mL distilled water, and water phase mixed solution is injected with 400uL/min in pear shape bottle solution,
700r/min aquations 15min under the conditions of 30 DEG C crosses the miillpore filter that aperture is 450nm, is centainly handled after being cooled to room temperature
After survey EE, experiment measures that the results are shown in table below.
Influence (n=3) of the ethanol consumption for EE
Ethanol consumption produces large effect for the EE of alcohol plastid, while with the increase of ethanol consumption, alcohol plastid
Grain size is also increasing, and many ruptures occur in the TEM photos that alcohol plastid is taken after placing when concentration of alcohol is excessive, inquires into one rationally
Ethanol consumption it is still necessary to want subsequent research.
The dosage of 2.3 β -4
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg addition 1mL ethanol solutions are placed in pear shape bottle, it will be a certain amount of
Extrasin beta -4 is dissolved in 1.75mL distilled water, water phase mixed solution is injected with 400uL/min in pear shape bottle solution, 30 DEG C of items
700r/min aquations 15min under part crosses the miillpore filter that aperture is 450nm after being cooled to room temperature, carry out surveying EE after centainly handling,
Experiment measures that the results are shown in table below：
Influence (n=3) of -4 dosage of extrasin beta for EE
- 4 dosage of extrasin beta produces strong influence, stabilizing influence of the dosage to alcohol plastid for the EE of alcohol plastid
It is larger.
The dosage of 2.4 surfactants
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in different amounts of ethanol solution and are placed in pear shape bottle,
0.50mg extrasin betas -4 and a certain amount of NaTDC are dissolved in 1.75mL distilled water, by water phase mixed solution with
400uL/min injects in pear shape bottle solution, 700r/min aquations 15min under the conditions of 30 DEG C, and aperture is crossed after being cooled to room temperature and is
The miillpore filter of 450nm carries out surveying EE after centainly handling, and experiment measures that the results are shown in table below.
Influence (n=3) of the dosage of surfactant for EE
The EE of alcohol plastid increases, anion surfactant energy with the increase of surfactant NaTDC dosage
Skin is penetrated into, is had an effect with iuntercellular lipoid, the mobility increase of lipid bilayer or lipid is caused to remove, is thin
Born of the same parents' peptide backbone structure changes, while alcohol plastid shelf-stability gradually increases, but zeta current potentials enhance simultaneously, for skin
Toxicity and irritation will increase, the electrification of general externally applied transdermal patch is no more than -65mv, so final NaTDC dosage is
6mg, alcohol plastid zeta current potentials are about -58mv or so at this time.
2.5 drop rate
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in the ethanol solution of 1mL and are placed in pear shape bottle, it will
0.50mg extrasin betas -4 are dissolved in 1.75mL distilled water, water phase mixed solution are injected with different rates in pear shape bottle solution, 30
700r/min aquations 15min under the conditions of DEG C crosses the miillpore filter that aperture is 450nm after being cooled to room temperature, carry out after centainly handling
EE is surveyed, experiment measures that the results are shown in table below：
Influence (n=3) of the rate of water injection for EE
With gradually slowing down for rate of water injection, aquation effect is more preferable, manufactured alcohol plastid encapsulation rate bigger.
2.6 aquation rotating speeds
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in the ethanol solution of 1mL and are placed in pear shape bottle, it will
0.50mg extrasin betas -4 and a certain amount of NaTDC are dissolved in 1.75mL distilled water, by water phase mixed solution with 400uL/
Min injects in pear shape bottle solution, and it is 450nm's that aperture is crossed with certain rotating speed aquation 15min under the conditions of 30 DEG C, after being cooled to room temperature
Miillpore filter carries out surveying EE after centainly handling, and experiment measures that the results are shown in table below.
Influence (n=3) of the aquation rotating speed for EE
Aquation rotating speed has a great impact for the formation of alcohol plastid and drugloading rate, in general, the faster aquation effect of rotating speed
Fruit is better, and encapsulation rate is preferable, but rotating speed is too fast to make alcohol plastid obtained largely rupture drug leakage, to make encapsulation rate
It is greatly reduced, TEM photos are shown in the alcohol plastid formed when stir speed (S.S.) is 500~900r/min and occur rupturing less, selection
700r/min does not occur alcohol plastid fracture phenomena substantially when being aquation rotating speed, therefore selects 700r/min for optimum speed.
2.7 hydration temperature
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in the ethanol solution of 1mL and are placed in pear shape bottle, it will
0.50mg extrasin betas -4 and a certain amount of NaTDC are dissolved in 1.75mL distilled water, by water phase mixed solution with 400uL/
Min injects in pear shape bottle solution, 700r/min aquations 15min under certain temperature, and it is the micro- of 450nm that aperture is crossed after being cooled to room temperature
Hole filter membrane carries out surveying EE after centainly handling, and experiment measures that the results are shown in table below.
Influence (n=3) of the hydration temperature for EE
From the data in the table, within the scope of 25-50 DEG C, hydration temperature is examined there is no the EE of significant impact alcohol plastid
Consider the too low and excessively high molding for being unfavorable for alcohol plastid of hydration temperature, therefore uses 30 DEG C of hydration temperature.2.8 hydration time
The cholesterol of 1.5mg Fabaceous Lecithins and 0.25mg are added in the ethanol solution of 1mL and are placed in pear shape bottle, it will
0.50mg extrasin betas -4 and a certain amount of NaTDC are dissolved in 1.75mL distilled water, by water phase mixed solution with 400uL/
Min injects in pear shape bottle solution, 700r/min aquations certain time under the conditions of 30 DEG C, and it is 450nm that aperture is crossed after being cooled to room temperature
Miillpore filter, carry out surveying EE after centainly handling, experiment measures that the results are shown in table below：
Influence (n=3) of the hydration time for EE
From the data in the table, there is no the EE of significant impact alcohol plastid for hydration time, it is contemplated that hydration temperature is too long
The stability of alcohol plastid may be destroyed, it is thus determined that hydration time is 10min.
3 orthogonal design optimization formulation of embodiment
Experiment of single factor in embodiment 2 the result shows that, to alcohol plastid be affected because being known as：The dosage (A) of cholesterol,
β -4 dosages (B), concentration of alcohol (C), drop rate (D), by orthogonal design L9 (3^4) table, the parameter evaluating as investigation is each
A prescription, following table are the investigation result of factor level table and orthogonal design respectively.
Orthogonal experiment factor and water-glass
Orthogonal experiment results
Determine that final prescription and technique are in conjunction with orthogonal test and experiment of single factor：By 1.5mg Fabaceous Lecithins and 0.25mg
During cholesterol addition 1mL ethanol solutions are set, 0.75mg extrasin betas -4 are dissolved in 1.75mL distilled water and are placed in pear shape bottle,
Alcohol phase mixed solution is injected with 200uL/min in pear shape bottle solution, 700r/min aquations 10min, is cooled under the conditions of 30 DEG C
The miillpore filter that aperture is 450nm is crossed after room temperature.
The preparation of 4 β -4 alcohol plastids of embodiment
(1) 0.02g Fabaceous Lecithins are added in 3mL absolute ethyl alcohols, are configured to phospholipid solution A, 0.02g cholesterol is added
In 1mL absolute ethyl alcohols, it is configured to cholesterol solution B；0.01g NaTDCs are added in the distilled water of 10mL and are configured to deoxidation
Sodium cholate solution C.
(2) it takes 0.225mL solution As, 0.5mL ethyl alcohol and 0.025mL solution B mixings in pear shape bottle, seals；
(3) 0.75mg extrasin betas -4 are dissolved in 1.125mL distilled water and the mixed solution of 0.625mL solution Cs；
(4) step (3) acquired solution is injected with 200uL/min in pear shape bottle solution, 700r/min water under the conditions of 30 DEG C
Change 10min, aperture is the miillpore filter of 450nm to get -4 alcohol plastid of extrasin beta excessively after being cooled to room temperature.As shown in Fig. 4,
It is 270 ± 3nm that obtained -4 alcohol plastid of extrasin beta measures grain size with Malvern dynamic light scattering particle size instrument；Such as 5 institute of attached drawing
Show, zeta current potentials are stablized in -58 ± 5mv；Its microscopic structures is as shown in 6 transmission electron microscope photo of attached drawing.
Utilize centrifugal determination encapsulation rate：Alcohol plastid will be made and be put in 10000r/min ultracentrifugations in ultracentrifuge
5min draws 20uL supernatants and measures absorbance at 562nm using microplate reader, establishes standard curve and calculates corresponding concentration.Packet
Envelope rate calculation formula is EE=(W2-W1)/W2*100%, and wherein w1 is β -4 contents of dissociating in alcohol plastid solution, and w2 is β -4
Total amount, w3 are the sum of the total amount of material and the total amount of β -4, and the encapsulation rate measured is 85 ± 5%.
The grain size and PDI of alcohol plastid prepared by 4 method of embodiment are measured, placement methods measure the stability of the alcohol plastid.
The preparation of 5 β -4 alcohol plastids of embodiment
(1) 0.02g phosphatide is added in 3mL ethanol solutions, is configured to phospholipid solution A, 0.02g cholesterol is added
In 1m ethanol solutions, it is configured to cholesterol solution B；0.01g NaTDCs are added in the distilled water of 10mL and are configured to
Sodium deoxycholate solution C.
(2) it takes 0.225mL solution As, 0.5mL ethyl alcohol and 0.029mL solution B mixings in pear shape bottle, seals
(3) 0.50mg extrasin betas -4 are dissolved in 1.59mL distilled water and 0.156mL solution Cs；
(4) mixed solution obtained by step (3) is injected with 200uL/min in pear shape bottle solution, 700r/ under the conditions of 30 DEG C
Min aquation 10min, aperture is the miillpore filter of 450nm to get -4 alcohol plastid of extrasin beta excessively after being cooled to room temperature.
The grain size and PDI of alcohol plastid prepared by 5 method of embodiment are measured, placement methods measure the stability of the alcohol plastid.
The preparation of 6 β -4 alcohol plastids of embodiment
(1) 0.02g phosphatide is added in 3mL absolute ethyl alcohols, is configured to phospholipid solution A, 1mL is added in 0.02g cholesterol
In absolute ethyl alcohol, it is configured to cholesterol solution B.
(2) it takes 0.225mL solution As, 0.5mL ethyl alcohol and 0.025mL solution B mixings in pear shape bottle, seals
(3) 0.75mg extrasin betas -4 are dissolved in 1.75mL distilled water；
(4) mixed solution obtained by step (3) is injected with 200uL/min in pear shape bottle solution, 700r/ under the conditions of 30 DEG C
Min aquation 10min, aperture is the miillpore filter of 450nm to get -4 alcohol plastid of extrasin beta excessively after being cooled to room temperature.
The grain size and PDI of alcohol plastid prepared by 6 method of embodiment are measured, placement methods measure the stability of the alcohol plastid.Through
Measurement and observation are crossed, the Comparative result of embodiment 4,5 and 6 is as shown in the table：
7 alcohol plastid physicochemical of embodiment
7.1 form
It takes appropriate PRO liposomes in cillin bottle, observes its appearance and light blue opalescence is presented.One to two drop alcohol plastids are taken again
Solution is suitably diluted with aquae destillata, is added dropwise on copper sheet, is dyed with 2% Salkowski's solution, is dried 30min, by transmiting electricity
The micro- sem observation microscopic pattern of son.As shown in Fig. 6, alcohol plastid form rounding under TEM, surface is smooth not to be adhered, and grain size is big
It is small to be evenly distributed, apparent fingerprint-like pattern can be observed, is that midparent water cavity is suitable for containing the big list of hydrophilic medicament greatly
Room alcohol plastid.
7.2 grain sizes and potential
It takes the liposome solutions prepared appropriate, is swashed using dynamic light scattering measurement Malvern after diluting suitable multiple
It is 270 ± 3nm that light particle size analyzer, which measures grain size,；Zeta current potentials are stablized in -58 ± 5mv.
7.3 alcohol plastid shelf-stabilities are investigated
Sample 1 and sample 2 is prepared according to 4 method of embodiment, investigates its 1d respectively, grain size and PDI when 10d, 20d,
As shown in the table：
Analysis：Alcohol plastid is after placing 10 days, 20 days.Grain size and PDI have good stability without substantially changeing.7.4β-4
The percutaneous abilities of alcohol plastid are investigated
Rat dorsum skin is taken to be clipped between diffusion cell lid (donor) and diffusion cell (acceptor), the inner surface of skin is heavy
It is immersed in isotonic solution physiological saline, 2mL β -4 alcohol plastids solution and free β -4 drugs is taken to be dipped in the physiological saline of 5mL respectively
In be used for transdermal diffusion.It measures 0.4mL solution from absorption cell in 1h, 2h, 4h, 6h, 8h, 10h, 12h respectively, and equivalent is added
Physiological saline.The unit area that the two is measured using microplate reader adds up transit dose (Q).
Experimental result：
Analysis：β -4 alcohol plastids are compared with the free drug of isoconcentration, and unit area adds up transit dose bigger, percutaneous abilities
More preferably.
8 alcohol plastid facial mask of embodiment
Gelling agent is widely used as a highly important exterior-applied formulation in percutaneous drug delivery, can be increased drug and be existed
The anelasticity of skin surface.Gel includes aqueous and oil-base gel agent, and alcohol plastid is a kind of waterborne liquid, so the present invention selects
Gel-type vehicle of one carbomer of gel as liposome with aqueous speciality.Such gel has good stretchability, oil-free
Greasy property has many advantages, such as skin good adhesion and without irritation, easy to clean, in the gel of D&C
It is widely used in agent.
8.1 preparation method
(1) preparation of alcohol plastid：Alcohol plastid is prepared according to the method described above；
(2) carbomer gel 934 and 940 for taking 0.5g, is placed in 24.5g distilled water, and magnetic agitation 400rpm/min is stirred
3.5h is mixed, stirring ends at left at room temperature over night, is fully swollen, then be slowly added into 0.125g freeze drying protectants (sucrose, grape
Sugar, mannitol) stir evenly Blank gel matrix to get water white transparency.The alcohol plastid solution prepared in advance is taken slowly to add
Enter into Blank gel matrix, grinds and uniformly to get β -4 Ethosomal gels.
The carbomer gel 934 and 940 of 0.5g is added in β -4 alcohol plastids in Example 4, is placed in 24.5g distilled water,
Magnetic agitation 400rpm/min stirs 3.5h, and stirring ends at left at room temperature over night, is fully swollen, then is slowly added into 0.125g jellies
Dry protective agent (sucrose, glucose, mannitol) stirs evenly the Blank gel matrix to get water white transparency.It takes and prepares in advance
Alcohol plastid solution be added slowly in Blank gel matrix, grind and uniformly to get β -4 Ethosomal gels.
As shown in fig. 7, β -4 Ethosomal gels uniform color obtained it is consistent, at translucent, the aobvious neutrality of acid-base value.It will
It is applied to rabbit skin of back 4h, after removing drug 30min, 1h, for 24 hours, 72h visually observe respectively, rabbit skin of back is not
Cause red and swollen or generates the adverse reactions such as macula.
8.2 appearance character
β -4 Ethosomal gels uniform color obtained is consistent, at translucent, the aobvious neutrality of acid-base value.It is applied to rabbit
Skin of back 4h, after removing drug 30min, 1h, for 24 hours, 72h visually observe respectively, rabbit skin of back do not cause it is red and swollen or
Generate the adverse reactions such as macula.
Above-mentioned, although the foregoing specific embodiments of the present invention is described with reference to the accompanying drawings, not protects model to the present invention
The limitation enclosed, those skilled in the art should understand that, based on the technical solutions of the present invention, those skilled in the art are not
Need to make the creative labor the various modifications or changes that can be made still within protection scope of the present invention.
Claims (10)
1. a kind of -4 alcohol plastid of extrasin beta, which is characterized in that be made of the raw material of following parts by weight：Extrasin beta -4
0.022%~0.043%, phosphatidase 0 .054%~0.084%, cholesterol 0.018%~0.028%, anionic surface activity
Agent 0.02%~0.04%, ethyl alcohol 10.10%~30.30%, remaining is distilled water.
2. alcohol plastid as described in claim 1, which is characterized in that the mass fraction of the raw material, which matches, is：Extrasin beta -4
0.03198%, phosphatidase 0 .06396%, cholesterol 0.02131%, anionic surfactant 0.02665%, ethyl alcohol
25.23%, remaining is distilled water.
3. -4 alcohol plastid of extrasin beta as described in claim 1, the anion surfactant is NaTDC or cholic acid
Sodium.
4. alcohol plastid as described in claim 1, which is characterized in that the phosphatide is Fabaceous Lecithin or lecithin, it is preferred that be beans
Phosphatide.
5. the preparation method of -4 alcohol plastid of extrasin beta described in claim 1, step include：
(1) phosphatide and cholesterol are dissolved in absolute ethyl alcohol acquisition alcohol phase solution, the phosphatide is Fabaceous Lecithin or lecithin, preferably
, it is Fabaceous Lecithin；
(2) anionic surfactant and extrasin beta -4 are dissolved in distilled water and are made into aqueous phase solution, the surfactant is
Anionic surfactant；Preferably, it is NaTDC or sodium taurocholate；
(3) aqueous phase solution is slowly injected into alcohol phase solution under stirring and carries out aquation；
(4) solution after aquation is cooled to room temperature, crosses miillpore filter to get -4 alcohol plastid of extrasin beta, the miillpore filter hole
Diameter is 450nm.
6. preparation method as claimed in claim 5, which is characterized in that ethanol solution concentration in mixed solution obtained by step (3)
It is 25%~40%.
7. preparation method as claimed in claim 5, which is characterized in that the rate stirred described in step (3) be 500~
800r/min。
8. preparation method as claimed in claim 5, which is characterized in that in step (3) water phase charge velocity be 0.2mL/min~
2mL/min。
9. preparation method as claimed in claim 5, which is characterized in that the time of aquation described in step (3) be 10~
30min, hydration temperature are 25~40 DEG C.
10. -4 alcohol plastid of extrasin beta as described in claim 1-4 is preparing the application for repairing facial mask field.
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| PCT/CN2018/108526 WO2020037783A1 (en) | 2018-08-21 | 2018-09-29 | THYMOSIN β-4 ETHOSOME AND PREPARATION PROCESS THEREOF |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114376975A (en) * | 2022-03-16 | 2022-04-22 | 南京星银药业集团有限公司 | Pharmaceutical composition ethosome, gel, hydrogel paste, patch and preparation method |
| CN115843864A (en) * | 2022-12-08 | 2023-03-28 | 江门职业技术学院 | Composite volatile oil ethosome and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102397255A (en) * | 2011-11-24 | 2012-04-04 | 广东药学院 | Progesterone ethosome, and preparation method and application thereof |
| CN105147689A (en) * | 2014-08-29 | 2015-12-16 | 中国中医科学院中药研究所 | Compound antimalarial composition |
| CN106975075A (en) * | 2016-01-18 | 2017-07-25 | 云南白药集团无锡药业有限公司 | Papain liposome gel Preparation method and use |
| CA3028721A1 (en) * | 2016-06-20 | 2017-12-28 | The Regents Of The University Of Michigan | Compositions and methods for delivery of biomacromolecule agents |
| CN108186438A (en) * | 2018-02-09 | 2018-06-22 | 广东科玮生物技术股份有限公司 | A kind of after-sun gel containing Porcine HGF and preparation method thereof |
-
2018
- 2018-08-21 CN CN201810951630.5A patent/CN108721604B/en active Active
- 2018-09-29 WO PCT/CN2018/108526 patent/WO2020037783A1/en active Application Filing
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102397255A (en) * | 2011-11-24 | 2012-04-04 | 广东药学院 | Progesterone ethosome, and preparation method and application thereof |
| CN105147689A (en) * | 2014-08-29 | 2015-12-16 | 中国中医科学院中药研究所 | Compound antimalarial composition |
| CN106975075A (en) * | 2016-01-18 | 2017-07-25 | 云南白药集团无锡药业有限公司 | Papain liposome gel Preparation method and use |
| CA3028721A1 (en) * | 2016-06-20 | 2017-12-28 | The Regents Of The University Of Michigan | Compositions and methods for delivery of biomacromolecule agents |
| CN108186438A (en) * | 2018-02-09 | 2018-06-22 | 广东科玮生物技术股份有限公司 | A kind of after-sun gel containing Porcine HGF and preparation method thereof |
Non-Patent Citations (5)
| Title |
|---|
| IBRAHIM M ABDULBAQI ET AL.: "Ethosomal Nanocarriers: The Impact of Constituents and Formulation Techniques on Ethosomal Properties, in Vivo Studies, and Clinical Trials", 《INT J NANOMEDICINE》 * |
| MAN MA ET AL.: "Development of nanovesicular systems for dermal imiquimod delivery: physicochemical characterization and in vitro/in vivo evaluation", 《J MATER SCI: MATER MED》 * |
| 廖立新 等: "胸腺素β4对人病理性瘢痕的影响", 《2013年全国中西医结合医学美容学术交流大会论文汇编》 * |
| 施晓琴 等: "醇质体在经皮给药方面的应用", 《中国实验方剂学杂志》 * |
| 朱金燕 等: "醇质体在经皮给药系统中的应用研究", 《包头医学院学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114376975A (en) * | 2022-03-16 | 2022-04-22 | 南京星银药业集团有限公司 | Pharmaceutical composition ethosome, gel, hydrogel paste, patch and preparation method |
| CN115843864A (en) * | 2022-12-08 | 2023-03-28 | 江门职业技术学院 | Composite volatile oil ethosome and preparation method thereof |
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| WO2020037783A1 (en) | 2020-02-27 |
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