CN105456194A - Magnolol liposome and derivative preparation and preparation method thereof - Google Patents
Magnolol liposome and derivative preparation and preparation method thereof Download PDFInfo
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- CN105456194A CN105456194A CN201510943994.5A CN201510943994A CN105456194A CN 105456194 A CN105456194 A CN 105456194A CN 201510943994 A CN201510943994 A CN 201510943994A CN 105456194 A CN105456194 A CN 105456194A
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- A—HUMAN NECESSITIES
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
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Abstract
The invention belongs to the field of medicine, and particularly relates to magnolol liposome and a derivative preparation and a preparation method thereof. The liposome prepared by taking magnolol as an active ingredient is capable of improving storage stability of pharmaceutic preparations as well as allowing drugs in the liposome to release sustainably. The magnolol liposome comprises, by weight, 1 part of magnolol, 2.5-11 parts of phospholipid, 0.4-3 parts of cholesterol and 0.10-3 parts of hydrophilic lipid derivatives. The preparation method of the liposome is simple and easy to implement, and the liposome is controllable in quality and convenient for industrial production; the magnolol liposome is small in particle size, uniform in distribution and high in entrapment efficiency, storage stability of the pharmaceutic preparations can be improved, the drugs in the liposome are allowed to release sustainably, retention time of the drugs in blood circulation is prolonged, bioavailability of the drugs is heightened, and the high-efficiency long-acting treatment goal is met.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to a kind of magnolol liposome and derivative preparation thereof and preparation method.
Background technology
Cortex Magnoliae Officinalis is the dry dried bark of China's Chinese herbal medicine Magnoliaceae magnolia Cortex Magnoliae Officinalis, root bark and branch skin.It is used to treat anxiety in Chinese medicine, cough, headache and anaphylactic disease.Due to the neuroprotective that it is good, be also used for the treatment of the mental sickness such as depression.Research shows, the hydroxylating biphenol compound magnolol (magnolol) separated from Cortex Magnoliae Officinalis has antiinflammatory, scavenging free radicals, alleviates traumatic brain injury, alleviates the effects such as acute lung injury.Also there are some researches show, magnolol antitumor, treatment pulmonary edema, prevention of arterial is atherosis, prevents skin photoage, suppress wrinkle formed, the aspects such as epilepsy also have certain effect.Although magnolol has physiologically active widely, magnolol is water insoluble, directly uses difficulty clinically, limits it and applies further.
Liposome (liposome) is that lipophilic drugs is wrapped in the miniature vesicle formed in lipoids bilayer by a class, effectively can improve the water solublity of lipophilic drugs, be widely used as the safe and effective carrier of hydrophilic and lipophilic drugs.Liposome can protect labile drug in human body, effectively transmits bioactive substance.Research shows, liposome not only has good safety, and has passive targeting, reduces the toxic and side effects of medicine while strengthening drug effect.In addition, the research that liposome stability improves becomes more effective pharmaceutical carrier.
Liposome, as pharmaceutical carrier, has certain targeting, especially not only to strengthen therapeutical effect but also can reduce drug toxicity with the combination of antitumor drug.Conventional liposome is made up of phospholipid and cholesterol, but targeting is poor, for improving guiding effect in the stability of liposome, targeting and cell, some functional group modified liposome surface can be adopted, as the liposome that polyglycol chain is modified, can prolong drug at blood circulation time, make liposome effectively can reach diseased region.
Liposome also has plurality of advantages as pharmaceutical carrier: the main component of (1) liposome is phospholipid and cholesterol, has biocompatibility, biodegradable, avirulence and reduced immunogenicity; (2) size, phospholipid composition, surface charge etc. of liposome have very large selection space; (3) lipid physical ability parcel hydrophilic and lipophilic drugs, enclose pharmaceutical pack in liposome, medicine can be protected not to be degraded in vivo, avoid medicine to the interference of Receptor recognition part simultaneously; (4) easily obtain suitable drug-carrier ratio, preparation is simple; (5) liposome can improve the curative effect of medicine and reduce its toxic and side effects; (6) different from solid polymer support system (as microgranule, nanoparticle), lipid film has good flexibility, and allow the targeted molecular of surface combination to have larger degree of freedom, therefore targeted molecular can with preferred configuration and target site receptors bind; (7) targeting that liposome can be institute's packaging medicine provides new possibility, comprises extracellular release, cell membrane fusion and endocytosis, improves the targeting scope of medicine; (8) be connected without covalent bond between liposome with medicine, be beneficial to medicine and discharge in lysosome; (9) particular target tissue and target cell can be delivered drugs at aglucon such as antibody, sugar ester etc. that drug-loaded liposome surface combination is different; (10) liposome is applicable to number of ways administration.
The present inventor for provide a kind of completely newly take magnolol as the liposome of active fraction preparation.
Summary of the invention
Technical problem solved by the invention is to provide a kind of stability that can improve pharmaceutical preparation and store, and the sustained drug in liposome can be made again to discharge with magnolol be the liposome of active fraction preparation.
Magnolol liposome of the present invention, the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidase 3-10 parts, 0.4 ~ 1 part, cholesterol, hydrophilic lipid derivate 0.10 ~ 0.25 part.
Preferably, magnolol 1 part, phosphatidase 3 .0 part, 0.4 part, cholesterol, hydrophilic lipid derivate 0.15 part.
Because parameters in commercial process occurs that the fluctuation of ± 10% is difficult to avoid, thus the prescription composition described in this patent and the higher limit of technological parameter or lower limit ± 10% scope in will think the protection domain of this patent.I.e. magnolol liposome of the present invention, the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidase 2 .5-11 part, 0.4 ~ 3 part, cholesterol, hydrophilic lipid derivate 0.10 ~ 3 part.
In technique scheme, described phospholipid can be at least one in natural phospholipid, semi-synthetic phospholipid, synthetic phospholipid or their derivant and above-mentioned substance hydrogenated products; Comprise at least one in soybean lecithin (SPC), Ovum Gallus domesticus Flavus lecithin (EPC), hydrogenated soy phosphatidyl choline (HSPC), hydrogenated yolk lecithin (HEPC), phosphatidylcholine (PC), PHOSPHATIDYL ETHANOLAMINE (PE), phosphatidyl glycerol (PG) and sodium salt thereof or phosphatidylinositols (PI), cuorin (cardiolipin, cardiolipin), sphingomyelin (SM), Phosphatidylserine (PS), phosphatidic acid (PA).
In technique scheme, described phosphatidylcholine (PC) is selected from but is not limited to distearoyl phosphatidylcholine (DSPC), dipalmitoyl phosphatidyl choline (DPPC), DOPC (DOPC), stearoyl oleoyl phosphatidylcholine, the sub-oleoyl phosphatidylcholine of stearoyl, POPC, the sub-oleoyl phosphatidylcholine of palmityl, dimyristoyl phosphatidyl choline (DMPC), DLPC (DLPC), DDPC, two decoyl phosphatidylcholines, at least one in DHPC or stearoyl palmitoylphosphatidylcholine.
In technique scheme, described PHOSPHATIDYL ETHANOLAMINE (PE) is selected from but is not limited to DSPE (DSPE), DPPE (DPPE), DOPE (DOPE), stearoyl oleoylphosphatidyl ethanolamine, the sub-oleoylphosphatidyl ethanolamine of stearoyl, palmitoyloleoyl phosphatidyl ethanolamine, the sub-oleoylphosphatidyl ethanolamine of palmityl, DMPEA (DMPE), two lauroyl PHOSPHATIDYL ETHANOLAMINE (DLPE), two caprinoyl PHOSPHATIDYL ETHANOLAMINE, at least one in two decoyl PHOSPHATIDYL ETHANOLAMINE or two hexanoyl PHOSPHATIDYL ETHANOLAMINE.
Further preferably, described phospholipid preferably adopts at least one in soybean lecithin (SPC), Ovum Gallus domesticus Flavus lecithin (EPC), hydrogenated yolk lecithin (HEPC), hydrogenated soy phosphatidyl choline (HSPC), distearoyl phosphatidylcholine (DSPC), dipalmitoyl phosphatidyl choline (DPPC), DSPE (DSPE), DPPE (DPPE).
In technique scheme, described hydrophilic lipid derivate is selected from polyethyleneglycol lipid derivates.Described polyethyleneglycol lipid derivates is selected from but is not limited to Polyethylene Glycol-sterol derivative as PEG-CHOL, PEG-CHOL succinate (PEG-CHEMS), PEG-CHOL methyl carbonate etc., polyethylene glycol-phosphorus lipid derivant is as PEG2000-DSPE (PEG-DSPE), Polyethylene Glycol-DPPE (PEG-DPPE), Polyethylene Glycol-DOPE, Polyethylene Glycol-palmitoyloleoyl phosphatidyl ethanolamine, Polyethylene Glycol-DMPEA etc., Polyethylene Glycol-Diglyceride or monoesters are as Polyethylene Glycol-distearyl glycerol, Polyethylene Glycol-dipalmitoyl-glycerol, Polyethylene Glycol-two myristoyl glycerol etc., Polyethylene Glycol-fatty acid or aliphatic ester derivatives etc. is as Polyethylene Glycol-stearic acid, Polyethylene Glycol-Palmic acid, Brij, at least one in Myrij.Wherein the molecular weight of Polyethylene Glycol be 100 ~ 20000, PEG and lipid part connecting key for but be not limited to disulfide bond, ester bond, ehter bond, amido link etc., PEG end group for but be not limited to hydroxyl, carboxyl, amino or nitrogen-containing group, alkoxyl etc.
Preferred further, described hydrophilic lipid derivate is selected from PEG2000-DSPE (PEG-DSPE), PEG-CHOL succinate (PEG-CHEMS), Monostalotetrahexosylgangliside (GM
1) or Tween 80 in one or more.
Preferred further, PEG2000-DSPE (PEG-DSPE) preferably adopts methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE (mPEG-DSPE).
To sum up, according to using methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE as optimum hydrophilic lipid derivate, then magnolol liposome of the present invention, the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidase 2 .5-11 part, 0.4 ~ 3 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 3 part;
Preferably, magnolol 1 part, phosphatidase 3-10 parts, 0.4 ~ 1 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 0.25 part;
Preferably, magnolol 1 part, phosphatidase 3 .0 part, 0.4 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.15 part.
To sum up, according to using methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE as optimum hydrophilic lipid derivate, phosphatidylcholine as optimum phospholipid, then magnolol liposome of the present invention, the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidylcholine 2.5-11 part, 0.4 ~ 3 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 3 part;
Preferably, magnolol 1 part, phosphatidylcholine 3-10 part, 0.4 ~ 1 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 0.25 part;
Preferably, magnolol 1 part, phosphatidylcholine 3.0 parts, 0.4 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.15 part.
The raw material of invented liposomes and consumption selection gist:
1) mass ratio controlling hydrophilic lipid derivate and phospholipid is 1:0.84 ~ 1:110, and preferred mass is than being 1:20 ~ 1:40, and the now less and even particle size distribution of the particle diameter of liposome, envelop rate is up to more than 90%, and preparation stability is good.
2) cholesterol in liposome has the effect of two-ways regulation phospholipid bilayer mobility, therefore the membrane stability of ratio regular meeting appreciable impact liposome of phospholipid and cholesterol and the rate of loading of medicine and degree.Preferably, in the prescription composition of magnolol liposome, the mass ratio of cholesterol and phospholipid is 1:0.84 ~ 1:27.5; The mass ratio of preferred cholesterol and phospholipid is 1:5 ~ 1:10, now less the and even particle size distribution of the particle diameter of liposome, and envelop rate is up to more than 90%, and preparation stability is good.
The preparation method of magnolol liposome of the present invention comprises: film dispersion method, injection method (alcohol injection, ether injection), ultrasonic dispersion.
In technique scheme, film dispersion method is also known as film evaporation method, and operational approach is fairly simple, is the earliest for one of the method for liposomal preparation.This method advantage is suitable for liposoluble drug delivery, and the entrapment efficiency of liposome of preparation is higher, and the Paclitaxel liposome of current domestic listing adopts this method to prepare and produces; This method shortcoming is with an organic solvent, and organic solvent eliminates more difficult, and comparatively consuming time, and suitability for industrialized production amplification ratio is more difficult.Comprise the following steps: the lipids such as phospholipid and the appropriate organic solvent of medicine (as chloroform) to be dissolved and after mix homogeneously, evaporate to dryness organic solvent, flask walls leaves a skim.Add buffer solution, water or normal saline, make film peel off from bottle wall.Stirring makes lipid film fully dissolve and be uniformly dispersed, and forms homogeneous dispersion liquid, obtained liposome.
In technique scheme, injection method is often referred to alcohol injection or ether injection.Alcohol injection operation is comparatively simple, comprises the following steps: medicine and lipid are first dissolved in alcoholic solution, is then injected into the buffer solution stirred at a terrific speed, in water or normal saline, after abundant mixing, use evaporate to dryness ethanol, obtained finely dispersed liposome.Ether injection comprises the following steps: phospholipid and cholesterol are dissolved in ether, adds magnolol alcoholic solution, shakes up.At the uniform velocity inject buffer solution again.After abundant mixing, boil off organic solvent, obtained liposome.The method has the advantages such as equipment is simple, cheap, easy to operate, liposome change in volume is little.
In technique scheme, ultrasonic dispersion comprises the following steps: water soluble drug is dissolved in phosphate buffer, adds phospholipid and cholesterol and fat-soluble medicine and is dissolved in altogether in the solution of organic solvent, stirs evaporating organic solvent, debris, through ultrasonic Treatment, then isolates liposome.
Magnolol liposome of the present invention both can directly apply to clinical as preparation, also can make derivative preparation as the raw material of other preparations.
Injection, lyophilized injectable powder, oral liquid, tablet, capsule, aerosol, nasal drop, gel etc. are comprised with the derivative preparation that magnolol liposome of the present invention is prepared for raw material.
Beneficial effect of the present invention:
Invented liposomes preparation method is simple, quality controllable, is convenient to suitability for industrialized production; The particle diameter of invented liposomes magnolol liposome little and distribute homogeneous, envelop rate is high, the stability that pharmaceutical preparation stores can be improved, the sustained drug in liposome can be made again to discharge, the holdup time of prolong drug in blood circulation, improve the bioavailability of medicine, reach efficient, long-acting therapeutic purposes.
Accompanying drawing explanation
Fig. 1 liposome solutions and magnolol aqueous solution outward appearance comparison diagram.
The comparison diagram of Fig. 2 magnolol liposome and magnolol solution release rate.
The Detection of Stability result of Fig. 3 magnolol liposome and blank liposome particle diameter.
The grain size distribution of Fig. 4 liposome
The Zeta potential figure of Fig. 5 liposome.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail; but these embodiments can not be understood to limit scope of the present invention; protection scope of the present invention limited by claims, and any change on the claims in the present invention basis is all protection scope of the present invention.
Embodiment one embodiment of the present invention adopts film dispersion method and alcohol injection to prepare liposome, investigates preparation method
1, alcohol injection is specifically implemented means and is: take magnolol, phosphatidylcholine, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE by recipe quantity precision, be dissolved in round-bottomed flask by absolute ethyl alcohol and stirring, be placed in water-bath heated at constant temperature, be stirred to adjuvant and be uniformly dissolved completely.Separately get 5ml ultra-pure water, heating, is vertically injected into alcoholic solution in the ultra-pure water of rapid stirring.Continue after injection completes to stir 10min, rotary evaporation removing dehydrated alcohol, standardize solution, to 5ml, obtains 2mg/ml liposome solutions.
2, the concrete enforcement means of film dispersion method are: take magnolol, phosphatidylcholine, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE by recipe quantity precision, be dissolved in organic solvent (chloroform) completely, be placed in round-bottomed flask, pressure reducing and steaming organic solvent, phospholipid is made to become translucent or white honeycomb film, by the abundant hydrated films of PBS, water-soluble ultrasonic a period of time, obtain liposome.
By the investigation and comparison to two kinds of methods, magnolol liposome prepared by film dispersion method dissolves slower in hydration process, and easily produce precipitation, also contain the shortcomings such as the larger organic solvent of the toxicity such as chloroform, it is simple that alcohol injection has equipment, cheap, the advantage such as easy to operate, therefore preferred alcohol injection method prepares liposome of the present invention.
The screening test of embodiment two magnolol liposome of the present invention formula
.1 the preparation of liposome
Groped by the test in early stage, tentatively establish the method preparing magnolol liposome with alcohol injection.Concrete grammar is: take magnolol, phosphatidylcholine, cholesterol, mPEG by recipe quantity analytical balance precision
2000-DSPE, is dissolved in round-bottomed flask by 2ml absolute ethyl alcohol and stirring, and be placed in water-bath and keep heated at constant temperature, magnetic agitation is uniformly dissolved completely to adjuvant, and this process used time is about 10-15min.Separately get 5ml ultra-pure water, heating, is injected into the alcoholic solution having dissolved adjuvant and medicine in the ultra-pure water of rapid stirring.Continue after injection completes to stir 5min, steam instrument rotary evaporation removing dehydrated alcohol with revolving, standardize solution, to 5ml, obtains the liposome solutions of 2mg/ml.Be placed into 4 DEG C of Refrigerator stores for subsequent use.
2, prescription screening
Be reference index through preliminary experiment with envelop rate, carry out single factor exploration to the factor that may affect magnolol liposome encapsulation, be optimized prescription.
The mensuration of 2.1 envelop rates
This experiment adopts dialysis to measure free magnolol and measures liposome encapsulation.Getting 1ml liposomal samples in molecular cut off is the bag filter of 12000 ~ 14000, dialyses in PBS solution, gets 0.5ml sample chromatograph methanol dilution to 1ml after dialysis 12h, and efficient liquid phase measures the content of free magnolol.Separately get 50 μ L magnolol liposome solutions, with chromatograph methanol dilution to 1ml, ultrasonic dissolution 30min, 13000rmin
-1centrifugal 40min, gets supernatant and measures magnolol total amount.1. liposome encapsulation is calculated by formula.
Envelop rate %=(amount of the total amount of magnolol-free magnolol)/magnolol total amount × 100%---formula 1.
2.2 single_factor method investigate mPEG
2000the addition of-DSPE is on the impact of liposome encapsulation
Table 1mPEG
2000the amount of-DSPE is on the impact of envelop rate
Measure envelop rate according to the formula preparation liposome in table 1.Result is known, and the liposome encapsulation be prepared into when the mass ratio of DSPE addition and magnolol is 1.5:10 is best.
The addition of 2.3 single factor exploration adjuvants is on the impact of liposome encapsulation
The amount of table 2 adjuvant is on the impact of envelop rate
The prescription screening of table 2 shows, liposome encapsulation obtained under 4 kinds of prescriptions, all more than 98%, illustrates that the enclose of liposome to magnolol is better.Consider envelop rate and save adjuvant two factors, when envelop rate is more or less the same, choosing the preparation that the 4th group of prescription saving adjuvant most carries out liposome.
2.4 single factor exploration results are known, and the preparation prescription of liposome should choose mPEG
2000the mass ratio of-DSPE addition and magnolol is 1.5:10, and the mass ratio of medicine and adjuvant is 10:30:4, i.e. magnolol: phospholipid: cholesterol: the preferred weight proportioning of hydrophilic lipid derivate is: 10:30:4:1.5.Sample appearance is have the good liposome solutions of blue-opalescent transparency, and after placing a few weeks longer, form has no obvious change.As Fig. 1, and directly with compared with water-soluble magnolol solution, after magnolol liposome, can effectively improve its dissolubility, be more prone to as medicine the utilization that is absorbed by the body.
3, extracorporeal releasing test and result thereof
Whether have slow release effect for investigating magnolol liposome, this experiment measures its release in vitro rate.
The preparation of 3.1 magnolol liposome honokiol solution
1) magnolol liposome is prepared by optimum formula preceding method.
2) take 12mg magnolol, use 3ml anhydrous alcohol solution, slowly add 3ml ultra-pure water, be mixed with 2mgmL
-1magnolol solution.
3.2 release experiment
Get magnolol liposome and each 1ml of magnolol solution, respectively get three parts and load bag filter, dialyse in 30mlPBS buffer, after dialysis starts, respectively sample 0.5ml in 0.5,1,2,4,6,8,12,24,48,72,96,120h, by chromatograph methanol constant volume to 1ml.Sample introduction successively, detect with HPLC and 2. calculate the release rate of magnolol and magnolol liposome with formula, its account form is: release rate %=dissociate magnolol amount/magnolol total amount × 100%---formula 2.
3.3 extracorporeal releasing experiment results
Draw release rate curve as Fig. 2.
As shown in Figure 2, rapidly, 0.5h has just discharged the medicine of more than 35% to the release of magnolol solution, liposome solutions just had same burst size after five days, can observe simultaneously and draw, in environment, magnolol solution discharges most medicine when dialysing 12h in vitro, reaches balance at 24h.It is comparatively slow that magnolol liposome solutions then discharges, until 120h is still at continuation slow releasing, learns that liposome really have slow release effect after contrasting with magnolol solution.
4 stability tests and result thereof
Preparation, in production application, except the factor considering cost and drug effect, also should consider the stability of medicine.This experiment, by the investigation prepared magnolol liposome encapsulation and particle diameter by optimum formula and carry out two weeks, evaluates the stability of this liposome.Concrete grammar is: magnolol liposomal samples is deposited in the refrigerator of 4 DEG C, measures its particle diameter respectively at 0,3,6,9,12,15d, draws stability curve as Fig. 3.
As shown in Figure 3, the particle diameter of magnolol liposome particle diameter within two weeks is stabilized in about 60nm.By contrast, the particle diameter fluctuation of blank liposome is comparatively large, and having good stability of magnolol liposome is described.
The mensuration of 5 particle diameters and Zeta potential
Magnolol liposome laser particle analyzer will be prepared carry out the detection of particle diameter and current potential by optimum formula.Result is as shown in Figure 4 and Figure 5: the mean diameter of liposome is 48.3 ~ 62.3nm, and polydispersity coefficient PDI is less than 0.3, illustrates that obtained liposomal particle size is evenly distributed.Zeta potential is-30.7mV, shows this liposome electronegative.Due to the electrostatic repulsion of molecular surface, when Zeta potential absolute value is more than or equal to 30mV, liposome stability is good.
Inventor is visible by above test, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE (mPEG
2000-DSPE) effect of finishing can be played in the preparation process of liposome, magnolol is more easily wrapping in liposome, increases the envelop rate of obtained magnolol liposome.And its content has material impact for magnolol liposome characteristics of pharmacokinetics in vivo and safety.Inventor has investigated mPEG
2000the addition of-DSPE, result shows, and the envelop rate of liposome affects in obviously experiment by it to be learnt: if do not add mPEG
2000-DSPE, obtained liposome solutions easily separates out precipitation, and blue-opalescent is not obvious and solution is transparent hardly, illustrates that a large amount of medicines is not liposomal encapsulated.Add mPEG
2000after-DSPE, obtained liposome character is the translucent solution with blue-opalescent, and placement was still stablized after two weeks.As can be seen here, mPEG is added when preparing liposome
2000-DSPE can significantly improve the envelop rate of magnolol liposome, and increase the stability of liposome, its optimal addn is mPEG
2000-DSPE and magnolol mass ratio are 1.5:10.
Illustrate that the present invention screens the beneficial effect of prescription scope below by way of preparation example.
The preparation and property of preparation example 1 invented liposomes detects:
Recipe quantity: magnolol, phospholipid, cholesterol and mPEG
2000the mass ratio of-DSPE is: 10:30:4:1.5
Prepared by employing alcohol injection: take magnolol, phosphatidylcholine, cholesterol, mPEG by analytical balance precision
2000-DSPE, is dissolved in round-bottomed flask by 2ml absolute ethyl alcohol and stirring, and be placed in water-bath heated at constant temperature, magnetic agitation is uniformly dissolved completely to adjuvant, and this process used time is about 10-15min.Separately get 5ml ultra-pure water, heating, by having dissolved, adjuvant is vertical with the alcoholic solution of medicine to be injected in the ultra-pure water of rapid stirring.Continue after injection completes to stir 5min, steam instrument rotary evaporation removing dehydrated alcohol with revolving, standardize solution, to 5ml, obtains the liposome solutions of 2mg/ml.Be placed into 4 DEG C of Refrigerator stores for subsequent use.
After testing, obtained magnolol liposome encapsulation 98.22%, particle diameter is about 60nm, and Zeta potential is about-30mV.Under 4 DEG C of conditions, within 15 days, lactone plastid has good stability.
The preparation and property of preparation example 2 invented liposomes detects:
Recipe quantity: magnolol, phospholipid, cholesterol and mPEG
2000the mass ratio of-DSPE is: 10:35:4:1.5
Prepared by employing alcohol injection: take magnolol, phosphatidylcholine, cholesterol, mPEG by analytical balance precision
2000-DSPE, is dissolved in round-bottomed flask by 2ml absolute ethyl alcohol and stirring, and be placed in water-bath heated at constant temperature, magnetic agitation is uniformly dissolved completely to adjuvant, and this process used time is about 10-15min.Separately get 5ml ultra-pure water, heating, is injected into the alcoholic solution having dissolved adjuvant and medicine in the ultra-pure water of rapid stirring.Continue after injection completes to stir 5min, steam instrument rotary evaporation removing dehydrated alcohol with revolving, standardize solution, to 5ml, obtains the liposome solutions of 2mg/ml.Be placed into 4 DEG C of Refrigerator stores for subsequent use.
Envelop rate 98.88%, particle diameter is about 70nm, and Zeta potential is about-28mV.Under 4 DEG C of conditions, within 15 days, lactone plastid has good stability.
The preparation and property of preparation example 3 invented liposomes detects:
Recipe quantity: magnolol, phospholipid, cholesterol and mPEG
2000the mass ratio of-DSPE is: 10:30:8:2.5
Prepared by employing alcohol injection: take magnolol, phosphatidylcholine, cholesterol, mPEG by analytical balance precision
2000-DSPE, is dissolved in round-bottomed flask by 2ml absolute ethyl alcohol and stirring, and be placed in water-bath heated at constant temperature, magnetic agitation is uniformly dissolved completely to adjuvant, and this process used time is about 10-15min.Separately get 5ml ultra-pure water, heating, is injected into the alcoholic solution having dissolved adjuvant and medicine in the ultra-pure water of rapid stirring.Continue after injection completes to stir 5min, steam instrument rotary evaporation removing dehydrated alcohol with revolving, standardize solution, to 5ml, obtains the liposome solutions of 2mg/ml.Be placed into 4 DEG C of Refrigerator stores for subsequent use.
Envelop rate 95.65%, particle diameter is about 75nm, and Zeta potential is about-30mV.Under 4 DEG C of conditions, within 15 days, lactone plastid has good stability.
From above-mentioned test, the present invention has successfully prepared magnolol liposome, and the parcel of the liposome developed by the present invention, the water solublity being insoluble in the magnolol of water is greatly improved.Preparation-obtained fat body envelop rate is high, and even particle size distribution, has good stability, and has slow releasing function, and be more easily absorbed by the body utilization, is better than prior art.
Claims (10)
1. magnolol liposome, is characterized in that: the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidase 2 .5-11 part, 0.4 ~ 3 part, cholesterol, hydrophilic lipid derivate 0.10 ~ 3 part;
Preferably, magnolol 1 part, phosphatidase 3-10 parts, 0.4 ~ 1 part, cholesterol, hydrophilic lipid derivate 0.10 ~ 0.25 part;
Preferably, magnolol 1 part, phosphatidase 3 .0 part, 0.4 part, cholesterol, hydrophilic lipid derivate 0.15 part.
2. magnolol liposome according to claim 1, is characterized in that: described phospholipid is at least one in natural phospholipid, semi-synthetic phospholipid, synthetic phospholipid or their derivant and above-mentioned substance hydrogenated products.
3. magnolol liposome according to claim 1, is characterized in that: described phospholipid comprises at least one in soybean lecithin, Ovum Gallus domesticus Flavus lecithin, hydrogenated soy phosphatidyl choline, hydrogenated yolk lecithin, phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE, phosphatidyl glycerol and sodium salt thereof or phosphatidylinositols, cuorin, sphingomyelin, Phosphatidylserine, phosphatidic acid.
4. magnolol liposome according to claim 3, is characterized in that: described phosphatidylcholine is at least one in distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, DOPC, stearoyl oleoyl phosphatidylcholine, the sub-oleoyl phosphatidylcholine of stearoyl, POPC, palmityl sub-oleoyl phosphatidylcholine, dimyristoyl phosphatidyl choline, DLPC, DDPC, two decoyl phosphatidylcholines, DHPC or stearoyl palmitoylphosphatidylcholine;
Described PHOSPHATIDYL ETHANOLAMINE is soybean phospholipid phosphatidyl ethanolamine, DSPE, DPPE, DOPE, stearoyl oleoylphosphatidyl ethanolamine, the sub-oleoylphosphatidyl ethanolamine of stearoyl, palmitoyloleoyl phosphatidyl ethanolamine, the sub-oleoylphosphatidyl ethanolamine of palmityl, DMPEA, two lauroyl PHOSPHATIDYL ETHANOLAMINE, two caprinoyl PHOSPHATIDYL ETHANOLAMINE, at least one in two decoyl PHOSPHATIDYL ETHANOLAMINE or two hexanoyl PHOSPHATIDYL ETHANOLAMINE.
5. magnolol liposome according to claim 1, is characterized in that: described phospholipid adopts at least one in soybean lecithin, Ovum Gallus domesticus Flavus lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline, distearoyl phosphatidylcholine, dipalmitoyl phosphatidyl choline, DSPE, DPPE.
6. magnolol liposome according to claim 1, is characterized in that: described hydrophilic lipid derivate is selected from polyethyleneglycol lipid derivates.
7. magnolol liposome according to claim 6, is characterized in that: described polyethyleneglycol lipid derivates comprises Polyethylene Glycol
1000 ~ 20000-sterol derivative, Polyethylene Glycol
1000 ~ 20000-fatty acid or aliphatic ester derivatives;
Further preferably, described Polyethylene Glycol
1000 ~ 20000-sterol derivative is Polyethylene Glycol
1000 ~ 20000-cholesterol, Polyethylene Glycol
1000 ~ 20000-cholesterol succinate, Polyethylene Glycol
1000 ~ 20000-cholesterol methyl carbonate etc., Polyethylene Glycol
1000 ~ 20000-phospholipid derivant is as Polyethylene Glycol
1000 ~ 20000-DSPE, Polyethylene Glycol
1000 ~ 20000-DPPE, Polyethylene Glycol
1000 ~ 20000-DOPE, Polyethylene Glycol
1000 ~ 20000-palmitoyloleoyl phosphatidyl ethanolamine, Polyethylene Glycol
1000 ~ 20000-DMPEA etc., Polyethylene Glycol
1000 ~ 20000-Diglyceride or monoesters are as Polyethylene Glycol
1000 ~ 20000-distearyl glycerol, Polyethylene Glycol
1000 ~ 20000-dipalmitoyl-glycerol, Polyethylene Glycol
1000 ~ 20000at least one in-two myristoyl glycerol;
Further preferably, described Polyethylene Glycol
1000 ~ 20000-fatty acid or aliphatic ester derivatives are Polyethylene Glycol
1000 ~ 20000-stearic acid, Polyethylene Glycol
1000 ~ 20000at least one in-Palmic acid, Brij, Myrij;
Preferably, described hydrophilic lipid derivate is selected from Polyethylene Glycol
1000 ~ 20000-DSPE, Polyethylene Glycol
1000 ~ 20000at least one in-cholesterol succinate, Monostalotetrahexosylgangliside or Tween 80.
8. magnolol liposome according to claim 1, is characterized in that: the component containing, for example lower weight proportion:
Magnolol 1 part, phosphatidase 2 .5-11 part, 0.4 ~ 3 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 3 part;
Preferably, magnolol 1 part, phosphatidase 3-10 parts, 0.4 ~ 1 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 0.25 part;
Preferably, magnolol 1 part, phosphatidase 3 .0 part, 0.4 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.15 part;
Preferably, magnolol 1 part, phosphatidylcholine 2.5-11 part, 0.4 ~ 3 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 3 part;
Preferably, magnolol 1 part, phosphatidylcholine 3-10 part, 0.4 ~ 1 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.10 ~ 0.25 part;
Preferably, magnolol 1 part, phosphatidylcholine 3.0 parts, 0.4 part, cholesterol, methoxy poly (ethylene glycol) PHOSPHATIDYL ETHANOLAMINE 0.15 part.
9. the preparation method of magnolol liposome described in any one of claim 1-8, is characterized in that comprising: film dispersion method, injection method, ultrasonic dispersion; Wherein, described injection method is alcohol injection, ether injection.
10. magnolol liposome derives preparation, it is characterized in that, with magnolol liposome described in any one of claim 1-8 for raw material, prepares injection, lyophilized injectable powder, oral liquid, tablet, capsule, aerosol, nasal drop, gel.
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| CN109875983A (en) * | 2018-12-10 | 2019-06-14 | 广州医科大学附属第二医院 | A kind of agonist of aldehyde dehydrogenase 2 |
| CN110974861B (en) * | 2019-12-24 | 2022-02-01 | 贵州大学 | Blumea balsamifera oil liposome |
| CN110974861A (en) * | 2019-12-24 | 2020-04-10 | 贵州大学 | Blumea balsamifera oil liposome |
| CN112121003A (en) * | 2020-09-29 | 2020-12-25 | 江苏集萃新型药物制剂技术研究所有限公司 | Drug-carrying material of sustained-release preparation, composition of drug-carrying material, sustained-release preparation and preparation method of sustained-release preparation |
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| CN115645383A (en) * | 2022-11-04 | 2023-01-31 | 武汉大学 | Application of magnolol in preparation of anti-EV 71 virus medicine |
| CN116617414A (en) * | 2023-03-31 | 2023-08-22 | 中国农业大学 | Liposome and preparation method and application thereof |
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| CN116270473A (en) * | 2023-05-25 | 2023-06-23 | 成都金瑞基业生物科技有限公司 | Co-carried liposome and preparation method thereof |
| CN116270473B (en) * | 2023-05-25 | 2023-12-19 | 成都金瑞基业生物科技有限公司 | Co-carried liposome and preparation method thereof |
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