CN102772802A - Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof - Google Patents
Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof Download PDFInfo
- Publication number
- CN102772802A CN102772802A CN201210228920XA CN201210228920A CN102772802A CN 102772802 A CN102772802 A CN 102772802A CN 201210228920X A CN201210228920X A CN 201210228920XA CN 201210228920 A CN201210228920 A CN 201210228920A CN 102772802 A CN102772802 A CN 102772802A
- Authority
- CN
- China
- Prior art keywords
- polyethylene glycol
- chitosan
- oleanolic acid
- nanometer liposome
- liposome
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicinal Preparation (AREA)
Abstract
The invention discloses a preparation method for an oleanolic acid (OA) nanoliposome modified by chitosan (CS) and polyethylene glycol (PEG). The method includes firstly, dissolving the OA, soybean phosphatidylcholines (SPC) and Chol into absolute ethyl alcohol, and stirring to obtain an oil phase; secondly, adding the PEG and a CS solution into a hydration medium phosphate buffer solution (PBS), and stirring to obtain a water phase; thirdly, dropwise adding the oil phase into the water phase to obtain a lipid suspension, fourthly, removing the absolute ethyl alcohol in the lipid suspension, and performing an ultrasonic dispersion to obtain a small monolayer liposome; and fifthly, preserving at the temperature of 4DEG C. An OA nanoliposome modified by the CS and the PEG is further disclosed. The nanoliposome is good in dispersity, neat in form, high in encapsulated ratio and capable of exerting effects of small scale and large absorbing surface, thereby the bioavailability of medicines can be remarkably improved, besides, the preparation of the OA nanoliposome can be completed under the normal pressure, normal temperature and mild conditions, the process is simple, the reaction is easy to control, and the encapsulated ratio of obtained liposome is high.
Description
Technical field
The invention belongs to the Nano medication technical field, oleanolic acid nanometer liposome of particularly a kind of chitosan and Polyethylene Glycol combined modification and preparation method thereof.
Background technology
OA (Oleanolic Acid, oleanolic acid) is a kind of natural pentacyclic triterpenoid, extensively is present in the various plants.OA is can transaminase lowering active, promote liver cell regeneration, blood sugar lowering, the fat relevant insulin resistance of alleviation and the effect of hyperlipidemia, and also has good antitumous effect.Because it does not have toxic and side effects to body, therefore has DEVELOPMENT PROSPECT preferably.But OA dissolubility in water is extremely low, absorbs in the body and some problems of biological utilisation existence, so its dosage is big, the medication number of times is frequent, and these have all limited giving full play to of its pharmacological action.
The absorption efficiency of medicine also depends on drug particles size, pattern, dispersibility and apparent condition etc. except receiving self property influences.Because it is too small that the influence of factors such as conventional medicament low absorption, metabolism and degraded makes medicine act on the concentration of diseased region, the drug solubility of water is low during administration, and these have all influenced medicine and have played a role at targeting moiety.Compare with the medicine of routine, Nano medication has that granule is little, surface reaction activity is high, the active center reaches advantages such as high adsorption capacity more.In addition, Nano medication can reduce dosage under the prerequisite that guarantees drug effect, alleviate or eliminate the toxic and side effects of medicine; For the relatively poor oral drugs of water solublity, reduce the drug particles size, its specific surface area be can increase, thereby the dissolution rate and the bioavailability of medicine significantly improved; The Nano medication that is wrapped can be realized its slow release effect, prolongs it at the intravital circulation time of machine, reduces administration number of times, improves drug effect and safety.
In the Nano medication system, liposome is one of at present best pharmaceutical carrier, is suitable for small-molecule drug, peptide, albumen and nucleic acid.It forms the transfer targeting through sealing, combine the protecting film or the blocking group that prevent biological elimination effect, thereby realizes nanoparticle target organ, target tissue or the desired selectivity of target cell.The administration of nanometer liposome oral administration can increase permeability and stability.It has higher drug loading, penetrates blood vessel easily and does not cause vascular endothelial injury, and the protection medicine is avoided the destruction of gastric acid and enzyme, and the local in vivo aggregate concentration of medicine is high, thereby can improve curative effect, and the reduction poisonous side effect of medicine; The nano-lipid physical ability increases its absorption at intestinal epithelial cell, prolongs soak time, improves drug targeting property.The method for preparing of liposome has a variety of, mainly contains active loading method and passive medicine carrying method, and active loading method comprises pH gradient method, ammonium sulphate gradient and calcium acetate gradient method; Passive medicine carrying method has film dispersion method, ultrasonic dispersion, freeze-drying, freeze-thaw method, multi-emulsion method, injection method, reverse evaporation, supercritical methanol technology.Because there are defectives such as poor stability, targeting property are not strong in common liposome, and the finishing of liposome has improved its targeting property, stability, realized the prolongation of action time in the body.At present, the method for modified liposome has chitosan and derivant, galactose, polyvinyl alcohol and different molecular weight polyethylene glycol (PEG).Chitosan is a kind of natural positively charged polysaccharide, characteristics such as its possess hydrophilic property, biocompatibility, biological degradability and hypotoxicity.Utilize chitosan modified liposome; Improve the slow-releasing of medicine, increased the stability of liposome and the targeting property of medicine, simultaneously can be through interacting with electronegative cell membrane; Promote the absorption of hydrophilic macromolecular drug, reduce the toxicity of cationic-liposome.Polyethylene Glycol and water molecules power are extremely strong, and flexible big, toxicity is low.The polyethyleneglycol modified circulating half-life of chemical compound of realizing prolongs and the physical and chemical stability enhancing.Can comprehensively bring into play both effects by chitosan and the common modified liposome film of PEG surface, make it reduce combining of conditioning ingredients in liposome and the blood plasma, increase hydrophilic, prolong its action time in vivo, improve its stability in vivo and in vitro.
But the liposome size for preparing through prior preparation method is big, dispersibility is bad, envelop rate is lower, thereby causes bioavailability of medicament low.
Summary of the invention
The technical problem that (one) will solve
The technical problem that the present invention will solve provides oleanolic acid nanometer liposome of a kind of chitosan and Polyethylene Glycol combined modification and preparation method thereof, with the low defective of bioavailability of the oleanolic acid liposome Chinese medicine that overcomes prior art.
(2) technical scheme
In order to achieve the above object; The invention provides the oleanolic acid nanometer liposome of a kind of chitosan and Polyethylene Glycol combined modification; Oleanolic acid is encapsulated between the duplicature of said nanometer liposome; And adopt chitosan and Polyethylene Glycol combined modification, and the mass ratio of said oleanolic acid and Polyethylene Glycol is 1:1.5, the mass ratio of said chitosan and Polyethylene Glycol is 1:1.5.
Wherein, said nanometer liposome is the spherical shape structure.
Wherein, said nanometer liposome size homogeneous.
The present invention also provides the method for preparing of the oleanolic acid nanometer liposome of a kind of chitosan and Polyethylene Glycol combined modification, said method comprising the steps of:
A, oleanolic acid, soybean lecithin and cholesterol are dissolved in the dehydrated alcohol, stir the oil phase that obtains homogeneous; The mass percent concentration of wherein said oleanolic acid and cholesterol is 1:2, and the mass ratio of said oleanolic acid and soybean lecithin is 1:10 ~ 1:20, and the volume of said dehydrated alcohol accounts for the 10-40% of said oil phase;
B, Polyethylene Glycol and chitosan solution joined in 35-45 ℃ the aquation medium phosphate buffer, stir and obtain water;
C, said oil phase is dropwise splashed into aqueous phase, obtain the lipid suspension;
D, remove the dehydrated alcohol in the said lipid suspension, and carry out Ultrasonic Pulverization and obtain small unilamellar vesicle;
E, place a period of time at normal temperatures, place 4 ℃ of preservations.
Wherein, in said steps A and step B, adopt the constant temperature magnetic agitation, wherein thermostat temperature is 35-55 ℃, and the rotating speed that in steps A, stirs is controlled at 300-700 r/min, and the rotating speed that in step B, stirs is controlled at 50-300 r/min.
Wherein, in said step B, the mass ratio of the oleanolic acid in said Polyethylene Glycol and the steps A is 1.5:1.
Wherein, in said step B, the pH value of said phosphate buffer is 6.5.
Wherein, in said step B, said chitosan solution is the solution that obtains in the acetic acid solution of 0.1-0.5% for chitosan is dissolved in concentration, and the mass ratio of said chitosan and Polyethylene Glycol is 1:1.5.
Wherein, in said step D, remove the method for the dehydrated alcohol in the said lipid suspension and remove or stir the time enough removal for the dehydrated alcohol in the lipid suspension being revolved boil off.
Wherein, in said step D, carry out the method that Ultrasonic Pulverization obtains small unilamellar vesicle and carry out the Ultrasonic Pulverization of 3-10 min for adopting the cell ultrasonic disintegrator.
(3) beneficial effect
The chitosan that obtains through method for preparing of the present invention and the oleanolic acid nanometer liposome good dispersion of Polyethylene Glycol combined modification, form is regular, envelop rate is higher; Can bring into play its small scale, big absorbing surface effect, thereby improve bioavailability of medicament more significantly.
In addition, the present invention can accomplish under the condition of normal temperature and pressure and gentleness, and technology is simple, and reaction is easy to control, and the liposome encapsulation that obtains is higher.
Description of drawings
Fig. 1 is the flow chart of method for preparing of oleanolic acid nanometer liposome of a kind of chitosan and the Polyethylene Glycol combined modification of the embodiment of the invention;
Fig. 2 a and Fig. 2 b are the oleanolic acid nanometer liposome electromicroscopic photograph figure of the modification of the embodiment of the invention;
Fig. 3 is that OA nanometer liposome and the former medicine of OA of CS of the present invention and PEG combined modification is to the survival rate of the Hela cell variable effect sketch map with drug level;
Fig. 4 is the infrared spectrum detection waveform figure of the embodiment of the invention to CS and PEG existence.
The specific embodiment
Below in conjunction with accompanying drawing and embodiment, specific embodiments of the invention describes in further detail.Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
The present invention adopts improved alcohol injection to combine ultrasonic dispersion; Investigated the formula factors (proportioning of OA and soybean lecithin; The proportioning of OA and ratio and the OA and the PEG-2000 of cholesterol) and preparation technology's factor (pH of hydration temperature in the preparation and aquation medium and ultrasonic time), select optimization formula through orthogonal experiment and single-factor variable method.The method for preparing of a kind of chitosan that the present invention is concrete and the oleanolic acid nanometer liposome of Polyethylene Glycol combined modification is as shown in Figure 1, may further comprise the steps:
Step s101 is dissolved in oleanolic acid, soybean lecithin and cholesterol in the dehydrated alcohol, stirs the oil phase that obtains homogeneous; The mass percent concentration of wherein said oleanolic acid and cholesterol is 1:2, and the mass ratio of said oleanolic acid and soybean lecithin is 1:10 ~ 1:20, and the volume of said dehydrated alcohol accounts for the 10-40% of said total system.Can adopt the constant temperature magnetic agitation in this step, thermostat temperature is 35-55 ℃, and rotating speed is controlled at 300-700 r/min.
Step s102 joins Polyethylene Glycol and chitosan solution in 35-45 ℃ the aquation medium phosphate buffer, stirs and obtains water.In this step; The mass ratio of the oleanolic acid among said Polyethylene Glycol and the step s101 is 1.5:1; The mass ratio of said chitosan and Polyethylene Glycol is 1:1.5, and the pH value of said phosphate buffer is 6.5, and adopts the constant temperature magnetic agitation; Thermostat temperature is 35-55 ℃, and rotating speed is controlled at 50-300 r/min.
Step s103 dropwise splashes into aqueous phase with said oil phase, obtains the lipid suspension.
Step s104 removes the dehydrated alcohol in the said lipid suspension, and carries out Ultrasonic Pulverization and obtain small unilamellar vesicle.In this step; The method of removing the dehydrated alcohol in the said lipid suspension is removed or is stirred time enough and remove for the dehydrated alcohol in the lipid suspension being revolved boil off, and carries out the method that Ultrasonic Pulverization obtains small unilamellar vesicle and carries out the Ultrasonic Pulverization of 3-10 min for adopting the cell ultrasonic disintegrator.
Step s105, room temperature place a period of time (general room temperature is placed and spent the night), place 4 ℃ of preservations.
The chitosan that the present invention obtains through above-mentioned method for preparing and the oleanolic acid nanometer liposome of Polyethylene Glycol combined modification are the spherical shape structure, and repeatability is high, and is cheap and easy to get; Prepared particle good dispersion, size is little, big or small homogeneous.Wherein oleanolic acid is encapsulated between the duplicature of said nanometer liposome, and the liposome encapsulation that obtains is higher.In the nanometer liposome of the present invention, the mass ratio of said oleanolic acid and Polyethylene Glycol is 1:1.5, and the mass ratio of said chitosan and Polyethylene Glycol is 1:1.5.
Embodiment one
In the present embodiment; OA, SPC (soybean lecithin) and Chol (cholesterol) are dissolved in a certain amount of dehydrated alcohol with the proportioning of 1:15:2,43 ℃ of constant temperature lower magnetic forces stir the oil phase that (rotating speed of stirring is 500 r/min) obtain homogeneous (wherein the volume of dehydrated alcohol account for total system 20%); PEG-2000 (mass ratio of OA and PEG is 1:1.5) and chitosan solution (0.1% acetic acid solution) join among 43 ℃ the aquation medium PBS (mass ratio of chitosan and PEG is 1:1.5); Stir (rotating speed of stirring is 120 r/min) at 43 ℃ of constant temperature lower magnetic forces and obtain water; The lipid alcoholic solution is dropwise splashed among the water PBS (phosphate buffer), get the lipid suspension.Dehydrated alcohol in the lipid suspension revolved to boil off remove or stir time enough and remove, adopt the cell ultrasonic disintegrator that its ultrasonic 7 min are obtained small unilamellar vesicle.The preparation back is subsequent use in 4 ℃ of preservations.
Wherein, oleanolic acid (OA) is available from Chengdu Man Site bio tech ltd; Soybean lecithin (SPC) is available from Shenyang Tian Feng Biology Pharmacy Co., Ltd; Cholesterol (Chol), chitosan (CS) is available from the sharp big biological Industrial Co., Ltd. in Zhengzhou; Polyethylene Glycol-2000 (PEG-2000) is available from the close europeanized reagent company limited product of Tianjin section.
Embodiment two
In the present embodiment; OA, SPC and Chol are dissolved in a certain amount of dehydrated alcohol with the proportioning of 1:10:2,55 ℃ of constant temperature lower magnetic forces stir the oil phase that (rotating speed of stirring is 300 r/min) obtain homogeneous (wherein the volume of dehydrated alcohol account for total system 10%); PEG-2000 (mass ratio of OA and PEG is 1:1.5) and chitosan solution (0.3% acetic acid solution) join among 45 ℃ the aquation medium PBS (mass ratio of chitosan and PEG is 1:1.5); Stir (rotating speed of stirring is 50 r/min) at 55 ℃ of constant temperature lower magnetic forces and obtain water; The lipid alcoholic solution is dropwise splashed among the water PBS, get the lipid suspension.Dehydrated alcohol in the lipid suspension revolved to boil off remove or stir time enough and remove, adopt the cell ultrasonic disintegrator that its ultrasonic 3 min are obtained small unilamellar vesicle.The preparation back is subsequent use in 4 ℃ of preservations.The raw material that adopts in the present embodiment is identical with embodiment one.
Embodiment three
In the present embodiment; OA, SPC and Chol are dissolved in a certain amount of dehydrated alcohol with the proportioning of 1:20:2,35 ℃ of constant temperature lower magnetic forces stir the oil phase that (rotating speed of stirring is 700 r/min) obtain homogeneous (wherein the volume of dehydrated alcohol account for total system 40%); PEG-2000 (mass ratio of OA and PEG is 1:1.5) and chitosan solution (0.5% acetic acid solution) join among 35 ℃ the aquation medium PBS (mass ratio of chitosan and PEG is 1:1.5); Stir (rotating speed of stirring is 300 r/min) at 35 ℃ of constant temperature lower magnetic forces and obtain water; The lipid alcoholic solution is dropwise splashed among the water PBS, get the lipid suspension.Dehydrated alcohol in the lipid suspension revolved to boil off remove or stir time enough and remove, adopt the cell ultrasonic disintegrator that its ultrasonic 10 min are obtained small unilamellar vesicle.The preparation back is subsequent use in 4 ℃ of preservations.The raw material that adopts in the present embodiment is identical with embodiment one.
Down in the face of measuring through the chitosan of the foregoing description acquisition and the oleanolic acid nanometer liposome of Polyethylene Glycol combined modification:
(1) sign of the OA nanometer liposome of CS and PEG combined modification: get nano liposome dispersion liquid under the room temperature and dilute, measure the mean diameter and the pattern of nanometer liposome at wavelength 215.0 nm (oleanolic acid characteristic absorption peak) with appropriate amount of deionized water.Get 1-2 and drip nano liposome dispersion liquid and place on the 300 purpose copper mesh, with 2% Salkowski's solution dyeing, room temperature is dried, and observes the form of nanometer liposome down in transmission electron microscope (TEM).
Test result: the oleanolic acid long-circulating nanoliposome microsphere good dispersion of finishing, narrow particle size distribution, below mean diameter 100 nm, the granule circularity of this liposome is very high, clear-cut, form is regular.The oleanolic acid nanometer liposome electromicroscopic photograph of modifying shown in Fig. 2 a and Fig. 2 b, the electromicroscopic photograph of Fig. 2 a when not adding the dosage form of Tween 80 wherein, Fig. 2 a electromicroscopic photograph during for the dosage form of adding Tween 80.
(2) entrapment efficiency determination of the OA nanometer liposome of dextrane gel chromatography CS and PEG combined modification: adopt the polydextran gel column chromatography to isolate liposome and free OA; The liposome effluent of collecting adopts the methanol breakdown of emulsion, and reversed-phase high-performance liquid chromatography (RP-HPLC) method is measured the drug level of sealing in medicine total concentration and the liposome.Under chromatographic condition, obtain the standard curve equation, and calculate the envelop rate of liposome medicament.
Test result: entrapment efficiency determination utilizes high performance liquid chromatograph, chromatographic column Angilent ZORBA * 300 SB-C18 (250 * 4.6mm, 5 μ m), and mobile phase is methanol: 0.1% acetic acid (88:12), UV-detector is made the oleanolic acid standard curve at 215nm.
Adopting the polydextran gel method to measure envelop rate, is filler with Sephadex G-75, and liposome and free medicine are separated, and finally is wrapped in the OA concentration (Ce) in the liposome with the standard curve Equation for Calculating.The accurate again post liposome 0.5ml that drew not puts the 10ml volumetric flask, uses the dissolve with methanol standardize solution, measures total drug level (Ct).
Calculate the envelop rate of liposome according to formula EE%=Ce/ Ct * 100%.Wherein, EE% is the envelop rate of liposome, and Ce is the OA concentration that is wrapped in the liposome, and Ct is the total concentration of OA in the Liposomal formulation.
(3) MTT (3-(4,5)-dimethylthiahiazo (z-y1)-3,5-di-phenytetrazoliumromide; 3-(4; 5-dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt) method detects the toxic action of the present invention to the Hela cell: the Hela cell of the trophophase of taking the logarithm is diluted to 5 * 103 cells/well density with it; Be inoculated in 96 orifice plates; Every hole, blank secondary series 6 holes adds each 100 μ l of DMEM culture medium, every hole, middle 58 holes 100 μ l Cell saps, and 32 holes, edge add the moisture of PBS with 64 holes, saturated centre.Administration behind the cultivation 24h, with 2,4,8,16,32 times of OA initial concentration dilutions, blank liposome dilutes identical multiple with LCL-OA simultaneously, aseptic filtration, each Concentraton gradient is done three parallel holes, the 10 μ l administrations of every hole.Behind the administration 24h, every hole adds MTT solution 10 μ l.After cultivating 4h in the incubator, every hole adds Formazan lysate 100 μ l, observes down until ordinary optical microscope and finds that purple crystal dissolves fully.Afterwards, adopt ELIASA under 595nm, to measure absorbance OD value.
The Hela cell that this test is adopted is provided by Beijing Medical University's Experimental Animal Center.The Hela cell is placed 37 ℃ of saturated humidities, contains 5%CO
2Incubator in cultivate.According to growing state 2-3 days of cell going down to posterity.When being exponential phase, cell is used for experiment.
Test result: adopt the fragmentation effect of the detection of drugs of mtt assay to tumor cell; The OD meansigma methods of the OA nanometer liposome of CS and PEG combined modification is starkly lower than the former medicine of OA; After administration 24h was described, the antitumous effect of the common OA nanometer liposome of modifying of CS and PEG was superior to the former medicine of OA.Wherein the OA nanometer liposome of CS and PEG combined modification and the former medicine of OA are as shown in Figure 3 with the variable effect of drug level to the survival rate of Hela cell.
(4) the present invention's detection that CS and PEG are existed: at first adopt mass percent concentration be mannitol and the sucrose of 1:1 as freeze drying protectant, use vacuum freeze drier that the OA liposome preparation of CS and PEG modification is become lyophilized powder.A spot of lyophilized powder and exsiccant KBr powder mixes are ground to form fine powder, and it is to be measured that the use tablet machine is pressed into thin slice.Its infrared spectrum detection waveform is as shown in Figure 4, and wherein, waveform A is pure PEG, and waveform B is pure CS, and waveform C is the OA liposome that PEG modifies, and waveform D is the common OA liposome of modifying of CS and PEG.As can beappreciated from fig. 4, the OA nanometer liposome of CS and PEG combined modification has the characteristic peak of CS and PEG-2000.
The embodiment of the invention adopts nanotechnology that oleanolic acid is prepared into the nanometer liposome of CS and PEG combined modification, the dissolubility that greatly improves medicine on the one hand and stability, improves the administering mode of oleanolic acid, reduces dosage, the raising therapeutic effect; Can bring into play its slow releasing function on the other hand.Experiment is observed its fragmentation effect to tumor cell through medicine of the present invention is carried out pharmacodynamic study.Simultaneously it has the multiple advantage that general Nano medication has as Nano medication, and as than small-size effect, specific surface area is big, and absorption efficiency is high, good stability etc.The method has simple to operate, and repeatability is high, the productive rate advantages of higher.
The present invention makes the nanometer liposome that Dispersion of Particles property is good, form is regular, envelop rate is higher through optimization of C factor and preparation technology's factor.The present invention is the film material with soybean lecithin and cholesterol, adopts improved alcohol injection to combine ultrasonic method to prepare the oleanolic acid nanometer liposome of chitosan and PEG combined modification.The liposome particles good dispersion that obtains, narrow particle size distribution, about mean diameter 100 nm, circularity is very high, clear-cut, form is regular.The present invention can effectively regulate and control the size of liposome particle diameter through the proportioning that changes film material in the lipid, and when the mass ratio of OA and SPC was 1:10, particle diameter was about 120 nm; During for 1:15, below particle diameter 200 nm; During for 1:20, particle diameter is more than 400 nm.Because the oleanolic acid long-circulating nanoliposome of finishing of the present invention has the obvious suppression effect to the Hela cell; And has certain slow release effect; The oleanolic acid of liposome as the carrier embedding is described; Can bring into play its small scale, big absorbing surface effect, thereby improve bioavailability of medicament more significantly.
In addition, the present invention can accomplish under the condition of normal temperature and pressure and gentleness, and technology is simple, and reaction is easy to control, and the liposome encapsulation that obtains is higher.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from know-why of the present invention; Can also make some improvement and replacement, these improvement and replacement also should be regarded as protection scope of the present invention.
Claims (10)
1. the oleanolic acid nanometer liposome of chitosan and Polyethylene Glycol combined modification; It is characterized in that; Oleanolic acid is encapsulated between the duplicature of said nanometer liposome; And adopt chitosan and Polyethylene Glycol combined modification, and the mass ratio of said oleanolic acid and Polyethylene Glycol is 1:1.5, the mass ratio of said chitosan and Polyethylene Glycol is 1:1.5.
2. the oleanolic acid nanometer liposome of chitosan according to claim 1 and Polyethylene Glycol combined modification is characterized in that, said nanometer liposome is the spherical shape structure.
3. the oleanolic acid nanometer liposome of chitosan according to claim 1 and 2 and Polyethylene Glycol combined modification is characterized in that, said nanometer liposome size homogeneous.
4. the method for preparing of the oleanolic acid nanometer liposome of chitosan and Polyethylene Glycol combined modification is characterized in that, said method comprising the steps of:
A, oleanolic acid, soybean lecithin and cholesterol are dissolved in the dehydrated alcohol, stir the oil phase that obtains homogeneous; The mass percent concentration of wherein said oleanolic acid and cholesterol is 1:2, and the mass ratio of said oleanolic acid and soybean lecithin is 1:10 ~ 1:20, and the volume of said dehydrated alcohol accounts for the 10-40% of said total system;
B, Polyethylene Glycol and chitosan solution joined in 35-45 ℃ the aquation medium phosphate buffer, stir and obtain water;
C, said oil phase is dropwise splashed into aqueous phase, obtain the lipid suspension;
D, remove the dehydrated alcohol in the said lipid suspension, and carry out Ultrasonic Pulverization and obtain small unilamellar vesicle;
E, place a period of time at normal temperatures, place 4 ℃ of preservations.
5. the method for preparing of the oleanolic acid nanometer liposome of chitosan according to claim 4 and Polyethylene Glycol combined modification; It is characterized in that; In said steps A and step B, adopt the constant temperature magnetic agitation, wherein thermostat temperature is 35-55 ℃; The rotating speed that in steps A, stirs is controlled at 300-700 r/min, and the rotating speed that in step B, stirs is controlled at 50-300 r/min.
6. the method for preparing of the oleanolic acid nanometer liposome of chitosan according to claim 4 and Polyethylene Glycol combined modification is characterized in that, in said step B, the mass ratio of the oleanolic acid in said Polyethylene Glycol and the steps A is 1.5:1.
7. according to the method for preparing of the oleanolic acid nanometer liposome of each described chitosan of claim 4 to 6 and Polyethylene Glycol combined modification, it is characterized in that in said step B, the pH value of said phosphate buffer is 6.5.
8. according to the method for preparing of the oleanolic acid nanometer liposome of each described chitosan of claim 4 to 6 and Polyethylene Glycol combined modification; It is characterized in that; In said step B; Said chitosan solution is the solution that obtains in the acetic acid solution of 0.1-0.5% for chitosan is dissolved in concentration, and the mass ratio of said chitosan and Polyethylene Glycol is 1:1.5.
9. the method for preparing of the oleanolic acid nanometer liposome of chitosan according to claim 4 and Polyethylene Glycol combined modification; It is characterized in that; In said step D, remove the method for the dehydrated alcohol in the said lipid suspension and remove or stir the time enough removal for the dehydrated alcohol in the lipid suspension being revolved boil off.
10. according to the method for preparing of the oleanolic acid nanometer liposome of claim 4 or 9 described chitosans and Polyethylene Glycol combined modification; It is characterized in that; In said step D, carry out the method that Ultrasonic Pulverization obtains small unilamellar vesicle and carry out the Ultrasonic Pulverization of 3-10 min for adopting the cell ultrasonic disintegrator.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210228920XA CN102772802A (en) | 2012-07-04 | 2012-07-04 | Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210228920XA CN102772802A (en) | 2012-07-04 | 2012-07-04 | Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102772802A true CN102772802A (en) | 2012-11-14 |
Family
ID=47118055
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210228920XA Pending CN102772802A (en) | 2012-07-04 | 2012-07-04 | Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102772802A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103919729A (en) * | 2014-03-20 | 2014-07-16 | 燕山大学 | Wedelolactone nanoliposome modified by combining chitosan and polyethylene glycol and its preparation method |
| CN107595779A (en) * | 2017-11-16 | 2018-01-19 | 河南牧翔动物药业有限公司 | A kind of rubican liposome and preparation method thereof |
| CN107714651A (en) * | 2017-11-16 | 2018-02-23 | 河南牧翔动物药业有限公司 | A kind of febantel liposome and preparation method thereof |
| CN111920957A (en) * | 2020-08-04 | 2020-11-13 | 河南中医药大学 | Preparation method of mitochondrion targeted tocopherol-oleanolic acid double-drug nanoparticles |
| CN112972403A (en) * | 2021-03-24 | 2021-06-18 | 齐鲁工业大学 | Lipid nano anti-tumor medicine containing bovine serum albumin and preparation method thereof |
| CN112999197A (en) * | 2021-03-05 | 2021-06-22 | 浙江医药高等专科学校 | Chitosan-coated solid lipid nanoparticle for promoting pentacyclic triterpenoid drug absorption and preparation method thereof |
| CN115869286A (en) * | 2022-11-10 | 2023-03-31 | 海南卓泰制药有限公司 | Amsacrine-containing encapsulating composition and preparation method thereof |
-
2012
- 2012-07-04 CN CN201210228920XA patent/CN102772802A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 吕青志等: "包覆脂质体的研究进展", 《食品与药品》 * |
| 高大威等: "齐墩果酸长循环脂质体的制备及表征", 《中国化学会第28届学术年会第3分会场摘要集》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103919729A (en) * | 2014-03-20 | 2014-07-16 | 燕山大学 | Wedelolactone nanoliposome modified by combining chitosan and polyethylene glycol and its preparation method |
| CN103919729B (en) * | 2014-03-20 | 2016-04-06 | 燕山大学 | The 1,8,9-trihydroxy-3-methoxy-benzo[4,5 nanometer liposome of a kind of chitosan and Polyethylene Glycol combined modification and preparation method |
| CN107595779A (en) * | 2017-11-16 | 2018-01-19 | 河南牧翔动物药业有限公司 | A kind of rubican liposome and preparation method thereof |
| CN107714651A (en) * | 2017-11-16 | 2018-02-23 | 河南牧翔动物药业有限公司 | A kind of febantel liposome and preparation method thereof |
| CN111920957A (en) * | 2020-08-04 | 2020-11-13 | 河南中医药大学 | Preparation method of mitochondrion targeted tocopherol-oleanolic acid double-drug nanoparticles |
| CN112999197A (en) * | 2021-03-05 | 2021-06-22 | 浙江医药高等专科学校 | Chitosan-coated solid lipid nanoparticle for promoting pentacyclic triterpenoid drug absorption and preparation method thereof |
| CN112972403A (en) * | 2021-03-24 | 2021-06-18 | 齐鲁工业大学 | Lipid nano anti-tumor medicine containing bovine serum albumin and preparation method thereof |
| CN112972403B (en) * | 2021-03-24 | 2022-03-29 | 齐鲁工业大学 | Lipid nano anti-tumor medicine containing bovine serum albumin and preparation method thereof |
| CN115869286A (en) * | 2022-11-10 | 2023-03-31 | 海南卓泰制药有限公司 | Amsacrine-containing encapsulating composition and preparation method thereof |
| CN115869286B (en) * | 2022-11-10 | 2023-08-18 | 海南卓泰制药有限公司 | Encapsulation composition containing amsacrine and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102772802A (en) | Oleanolic acid nanoliposome modified by chitosan and polyethylene glycol and preparation method thereof | |
| CN103705469B (en) | A kind of honokiol nanoparticle and preparation method thereof | |
| US20190224238A1 (en) | Tumor therapeutic drug | |
| CN114099533A (en) | Nucleic acid drug delivery system, preparation method, pharmaceutical composition and application | |
| CN102686217B (en) | Submicro emulsion of paclitaxel using steroid complex as intermediate carrier | |
| CN101953792B (en) | Irinotecan nano circulating liposome and preparation method thereof | |
| CN101984958B (en) | Nanoscale albendazole micropowder and preparation method thereof | |
| CN109288794A (en) | A kind of melittin liposome nanometer formulation and the preparation method and application thereof | |
| Guo et al. | High oral bioavailability of 2-methoxyestradiol in PEG-PLGA micelles-microspheres for cancer therapy | |
| CN112022919A (en) | Percutaneous-absorption artemisia vulgaris oil carrier gel and preparation method thereof | |
| CN102552182A (en) | Colloidal nucleus liposome lyophilized powder and preparation method thereof | |
| Han et al. | In vitro and in vivo evaluation of core–shell mesoporous silica as a promising water-insoluble drug delivery system: Improving the dissolution rate and bioavailability of celecoxib with needle-like crystallinity | |
| CN104887630A (en) | A lipidosome-covered dehydrogenated silibinin phospholipid complex and a preparing method thereof | |
| CN104288093A (en) | Application of nano-drug transdermal preparation in tumors | |
| CN102397242A (en) | Hydrogel containing coenzyme Q10, and cataplasm prepared by hydrogel | |
| CN105031658A (en) | Preparation method for acidity-controllable drug carrier | |
| CN102657610A (en) | 3,5-dihydroxyl4-isopropyl diphenylethylene micro-emulsion and preparation method thereof | |
| CN102379850A (en) | Targeted administration liposome passing through mucus barriers of human bodies | |
| CN102188378B (en) | Preparation method of liposome for coating and carrying water soluble drugs | |
| Shakhova et al. | Niosomes: a promising drug delivery system | |
| CN102973525B (en) | Antharcycline antitumor antibiotics loaded nano-micelle preparation and preparation method thereof | |
| CN105287612A (en) | Salinomycin sodium and adriamycin co-loaded nano-liposome as well as preparation method and application thereof | |
| Hu et al. | Anticancer effect of folic acid modified tumor-targeting quercetin lipid nanoparticle | |
| CN102188379A (en) | Preparation method of drug-carrying liposome | |
| CN103006560B (en) | Hyaluronic acid oligosaccharide encased paclitaxel liposome and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121114 |