CN101683322B - Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method - Google Patents
Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method Download PDFInfo
- Publication number
- CN101683322B CN101683322B CN2008102005466A CN200810200546A CN101683322B CN 101683322 B CN101683322 B CN 101683322B CN 2008102005466 A CN2008102005466 A CN 2008102005466A CN 200810200546 A CN200810200546 A CN 200810200546A CN 101683322 B CN101683322 B CN 101683322B
- Authority
- CN
- China
- Prior art keywords
- carbon dioxide
- berberine hydrochloride
- supercritical carbon
- liposome
- hydrochloride 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a method for preparing nano berberine hydrochloride liposome by a supercritical carbon dioxide method. The invention has the following specific operation steps: 1) forming a film; 2) encapsulating drugs; a) preparing berberine hydrochloride liposome injectable suspension; b) incubating in supercritical carbon dioxide fluid; and 3) after finishing releasing the carbon dioxide incubation, releasing carbon dioxide gas and reducing the system pressure into normal pressure to obtain the product. Compared with the prior art, the invention adopts the supercritical carbon dioxide fluid as a medium to prepare the nano berberine hydrochloride liposome with good monodispersity and high drug encapsulation rate according to the characteristics of even mass transferring and high velocity and the like of the supercritical carbon dioxide fluid, and controls the dimension of the liposome by regulating and preparing pressure.
Description
Technical field
The invention belongs to and relate to the pharmaceutical carrier preparation field, specifically a kind of method for preparing of nano berberine hydrochloride liposome.
Background technology
After medicine combined with carrier and forms medicament carrier system, the absorption of medicine and distributing no longer determined by medicine itself, but received the influence of carrier physicochemical property.Select the suitable pharmaceutical carrier material according to clinical requirement, not only can conduct drugs to target organ, and play useful effect for the physicochemical property and the pharmacologically active of medicine.
Berberine hydrochloride (berberine) is a kind of isoquinoline alkaloid that from plant drying rhizomes such as Rhizoma Coptidis, Cortex Phellodendri, Radix Berberidis, extracts, and is mainly used in the past through intestinal canal administration and treats gastroenteritis, bacillary dysentery and general intestinal infection etc.In recent years discover berberine hydrochloride to the human liver cancer cell, colon tumor cell, esophageal cancer cell system, malignant teratoma cell, rat 9L brain tumor cell, Lewis lung cancer cell etc. all has certain inhibition and killing action.But research finds that also tumor-inhibiting action does not have interaction in vitro obvious in the berberine hydrochloride body, possibly be because the berberine hydrochloride intestinal absorption is relatively poor, is difficult in vivo reach to the direct acting higher blood drug level of tumor cell.The berberine hydrochloride intravenously administrable possibly cause side effect such as drug eruption, anaphylaxis, hematochrome and erythrocytopenia, drug sensitivity shock, and nineteen eighty-two, China banned use of the berberine hydrochloride intravenously administrable.Because berberine hydrochloride has cheap, safe characteristics, make it become a kind of medicine that is worth further investigation aspect treatment and prevention of tumour.Therefore, how to bring into play the effect of berberine hydrochloride in treatment of cancer, suppressing its side effect is the big problem that we face.In order to improve the berberine hydrochloride curative effect, reduce its toxic and side effects simultaneously, this patent proposes the new method of preparation nano berberine hydrochloride liposome.
The superminiature spheroid that liposome type of being lipid bilayer forms, similar biofilm structure, it can be wrapped in pharmaceutical pack in the capsule and become a kind of new drug carrier.Its solvent phospholipid is inherent composition in the organism, degrades through biotransformation in vivo, and avirulence and immunogenicity, therefore, liposome is considered to one of pharmaceutical carrier of tool development prospect.
At present, the method for preparing liposome mainly contains film dispersion method, reverse evaporation, ultrasonic dispersing method; Freeze-thaw method, freeze-drying, alcohol injection, ether injection method; Reverse phase evaporation, surfactant treatment method etc., but there is important disadvantages in above-mentioned method for preparing; As: organic solvent residual causes the safety of liposome to descend in the product, and granule monodispersity difference is unfavorable for the stable of system, and the envelop rate of medicine is low etc.Supercritical carbon dioxide fluid is a kind of environmentally friendly solvent, after preparation finishes, is easy to separate with product and do not cause environmental pollution, and possesses the characteristic of fine solvent.In recent years, along with the supercritical carbon dioxide fluid technology rapid development, the continuous expansion of application, it is applied in the preparation of liposome gradually.In the liposome preparation process, the advantage of supercritical carbon dioxide mainly shows the following aspects: the first, and with other method for preparing comparison of liposome, the liposome particle diameter of supercritical carbon dioxide process preparation is comparatively even, and monodispersity is high.The second, with operation such as evaporation, extraction or dilution relatively, utilize the liposome organic solvent-free of supercritical carbon dioxide process preparation residual, for the safe clinical practice of liposome provides maybe.The 3rd, in hatching process, supercritical carbon dioxide fluid can dissolve the small molecular weight impurity in the phospholipid, and impurity can be simultaneously by the stripped system of band, to reach the effect that purification phospholipid improves the liposome quality during release of carbon dioxide.The 4th; Supercritical carbon dioxide fluid is a kind of emerging fine solvent; Mass transfer rate is high, and this helps the diffusion of hatching process Chinese medicine molecule in the export-oriented film of film, and supercritical carbon dioxide fluid is that carbon dioxide forms through the high pressure effect at a certain temperature simultaneously; High pressure in the system helps the interior water that medicine gets into liposome equally, and then improves the envelop rate of liposome.In addition, the supercritical carbon dioxide fluid environment can plastify the structure of phospholipid molecule, and this plastication helps phospholipid molecule and forms the more stable conformation of thermodynamics, makes the arrangement of bilayer more even, to reach the purpose that improves liposome stability.Above-mentioned advantage explanation supercritical carbon dioxide process has certain advantage aspect the preparation berberine hydrochloride liposome.
Summary of the invention
In the method to the existing preparation liposome in the ubiquitous product organic solvent residual cause the safety of liposome to descend; Granule monodispersity difference is unfavorable for defectives such as stablizing of system; The present invention proposes a kind of new method of preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method, and particle diameter in the traditional preparation process method is inhomogeneous to overcome effectively, poor stability, envelop rate are low, the shortcoming of organic solvent residual.
The technical scheme that the present invention adopts is:
Mass transfer according to supercritical carbon dioxide fluid evenly reaches characteristics such as speed height; The utilization supercritical carbon dioxide fluid is as medium; The preparation nano berberine hydrochloride liposome that monodispersity is good, entrapment efficiency is high, and can be through regulating the size of preparation pressure control liposome.
According to technique scheme, concrete operations of the present invention are:
1) film forming
Take by weighing an amount of lecithin and cholesterol and place round-bottomed flask, the weight ratio of lecithin and cholesterol is 3.8-2.5:1.With the chloroform methanol mixed liquor of volume ratio 2.5-2:1 preparation, add again wherein extremely with till its dissolving.After dissolving fully, under 35-42 ℃ of constant temperature and nitrogen protection condition, film forming in Rotary Evaporators, evacuation volatilizees organic solvent fully then.
2) entrapped drug
A) preparation of berberine hydrochloride liposome suspension
Pipette the berberine hydrochloride drug solution, the round-bottomed flask that places step 1) to have, magnetic agitation comes off film, obtains the berberine hydrochloride liposome suspension.Wherein the mass ratio of berberine hydrochloride drug solution and lecithin (medicine fat ratio) is 1:9-12.
B) in supercritical carbon dioxide fluid, hatch
The suspension that makes in a) is placed autoclave, and other adds an amount of dehydrated alcohol, again autoclave is placed water bath with thermostatic control; Feed carbon dioxide; After reaching uniform temperature and pressure, carbon dioxide gets into its critical region, presents the supercritical fluid state.Preparation pressure can be regulated.As 16,18,20,22,24MPa etc.As long as this pressure carbon dioxide is above-critical state.Close intake valve then, 40-45 ℃ stirred in water bath hatching half an hour.The mass ratio of dehydrated alcohol and berberine hydrochloride drug solution is 1:4-5.5.Getting dehydrated alcohol density is 0.789g/mL, and the density of berberine hydrochloride drug solution is approximately 1g/mL.
3) release of carbon dioxide
After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, promptly gets product.
Compare prior art, the present invention has following good effect:
The particle size dispersion of the berberine hydrochloride liposome that 1) prepares is even, and monodispersity is good, stability high (Fig. 1, Fig. 2, table 1);
2) visible like the comparison of table 2, the envelop rate of the medicine of gained of the present invention is higher than traditional method;
3) Determination of Residual Organic Solvents is few;
4) can be when regulating hatching the size and the stability (seal pressure to size and dispersive influence thereof, see Fig. 3) of the pressure control liposome particle diameter of supercritical carbon dioxide in the autoclave.The relation of preparation pressure and berberine hydrochloride liposome particle diameter: pressure is big more, and particle diameter is more little.
Description of drawings
Fig. 1 is the particle diameter and the form (AFM) of nano berberine hydrochloride liposome
Fig. 2 a and Fig. 2 b are the size ratios (laser light scattering mensuration result) of the berberine hydrochloride liposome of the method for preparing that proposes of the present invention and traditional film dispersion method preparation.
Wherein Fig. 2 a is particle diameter and the particle size distribution that supercritical carbon dioxide process that the present invention proposes prepares nanometer liposome, and Fig. 2 b is the particle diameter and the particle size distribution of the liposome of film dispersion method preparation.
Fig. 3 shows through regulating the preparation pressure of supercritical carbon dioxide, can control the size and the dispersibility of nano berberine hydrochloride liposome
Embodiment
Below further specify the present invention through embodiment:
Embodiment 1: preparation berberine hydrochloride liposome suspension; Take by weighing 50mg lecithin and the 16.5mg cholesterol places round-bottomed flask; Chloroform and methyl alcohol mixed liquor dissolving with 3mL volume ratio 2.2:1; Dissolve back rotary evaporation film forming fully, bath temperature is set at 41 ℃, and evacuation makes solvent evaporates again.Pipette 6ml1mg/mL berberine hydrochloride solution, place above-mentioned round-bottomed flask, magnetic agitation comes off film.Above-mentioned suspension and magneton are placed autoclave, and other adds the 1.3mL dehydrated alcohol, and logical carbon dioxide is to 12MPa.Close intake valve then, in the stirred in water bath hatching half an hour of uniform temperature.After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, promptly gets product.
Embodiment 2: preparation berberine hydrochloride liposome suspension; Take by weighing 50mg lecithin and the 16.5mg cholesterol places round-bottomed flask, with chloroform and the methyl alcohol mixed liquor dissolving of 4mL volume ratio 2:1, dissolving back rotary evaporation film forming fully; Bath temperature is set at 45 ℃, and evacuation makes solvent evaporates again.Pipette 5mL1mg/mL berberine hydrochloride solution, place above-mentioned round-bottomed flask, magnetic agitation comes off film.Above-mentioned suspension and magneton are placed autoclave, and other adds the 1.3mL dehydrated alcohol, and logical carbon dioxide is to 16MPa.Close intake valve then, in the stirred in water bath hatching half an hour of uniform temperature.After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, promptly gets product.
Embodiment 3: preparation berberine hydrochloride liposome suspension; Take by weighing 50mg lecithin and the 16.5mg cholesterol places round-bottomed flask; Chloroform and methyl alcohol mixed liquor dissolving with 3.5mL volume ratio 2.5:1; Dissolve back rotary evaporation film forming fully, bath temperature is set at 42 ℃, and evacuation makes solvent evaporates again.Pipette 5mL1mg/mL berberine hydrochloride solution, place film forming round-bottomed flask, magnetic agitation comes off film.Above-mentioned suspension and magneton are placed autoclave, and other adds the 1.3mL dehydrated alcohol, and logical carbon dioxide is to 20MPa.Close intake valve then, in the stirred in water bath hatching half an hour of uniform temperature.After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, promptly gets product.
Embodiment 4: preparation berberine hydrochloride liposome suspension; Take by weighing 55mg lecithin and the 16.5mg cholesterol places round-bottomed flask, with chloroform and the methyl alcohol mixed liquor dissolving of volume ratio 2.1:1, dissolving back rotary evaporation film forming fully; Bath temperature is set at 43 ℃, and evacuation makes solvent evaporates again.Pipette 5mL1mg/mL berberine hydrochloride solution, place above-mentioned round-bottomed flask, magnetic agitation comes off film.Above-mentioned suspension and magneton are placed autoclave, and other adds the 1.3mL dehydrated alcohol, and logical carbon dioxide is to 24MPa.Close intake valve then, in the stirred in water bath hatching half an hour of uniform temperature.After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, promptly gets product.
The present invention evenly reaches characteristics such as speed height according to the mass transfer of supercritical carbon dioxide fluid; The utilization supercritical carbon dioxide fluid is as medium; Preparation monodispersity is good, entrapment efficiency is high nano berberine hydrochloride liposome, and can be when regulating hatching the size of the pressure control liposome particle diameter of supercritical carbon dioxide in the autoclave.
The liposome stability of supercritical carbon dioxide process that table 1, the present invention propose and the preparation of traditional supercritical carbon dioxide process relatively
(annotate: 1, the condition determination of stability constant: a calculation of Stability Constants formula: K
E=(A
25d-A
0)/A
0The b temperature is 277.2K;
2, stability constant small liposome is stable more more)
The liposome encapsulation of supercritical carbon dioxide process that table 2, the present invention propose and the preparation of traditional supercritical carbon dioxide process relatively
(annotate: the computing formula of envelop rate: envelop rate=(1-free drug quality/medicine gross mass) * 100%)
Claims (6)
1. the method for a preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method is characterized in that it may further comprise the steps:
1) film forming
Take by weighing an amount of lecithin and cholesterol and place round-bottomed flask, the weight ratio of lecithin and cholesterol is 3.8-2.5: 1, and again with volume ratio 2.5-2: the chloroform methanol mixed liquor of 1 preparation adds wherein extremely with till its dissolving; After dissolving fully, under 35-42 ℃ of constant temperature and nitrogen protection condition, film forming in Rotary Evaporators, evacuation volatilizees organic solvent fully then;
2) entrapped drug
A) preparation of berberine hydrochloride liposome suspension
Pipette the berberine hydrochloride drug solution, the round-bottomed flask that places step 1) to have, magnetic agitation comes off film, obtains the berberine hydrochloride liposome suspension; The mass ratio of described berberine hydrochloride drug solution and lecithin is 1: 9-12;
B) in supercritical carbon dioxide fluid, hatch
The suspension that makes in a) is placed autoclave, and other adds an amount of dehydrated alcohol, again autoclave is placed water bath with thermostatic control, feeds carbon dioxide, and after the increase pressure, carbon dioxide gets into its critical region, presents the supercritical fluid state; Close intake valve then, 40-45 ℃ stirred in water bath hatching 20-45 minute; The mass ratio of dehydrated alcohol and berberine hydrochloride drug solution is 1: 4-5.5;
3) release of carbon dioxide
After hatching finished, carbon dioxide gas was reduced to normal pressure with system pressure, obtains product.
2. the method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method as claimed in claim 1 is characterized in that the mass ratio of described berberine hydrochloride drug solution and lecithin is preferably 1: 10.
3. the method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method as claimed in claim 1 is characterized in that the mass ratio of described dehydrated alcohol and berberine hydrochloride drug solution is preferably 1: 5.
4. the method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method as claimed in claim 1, when it is characterized in that in supercritical carbon dioxide fluid, hatching, the temperature of water-bath is preferably 43 ℃.
5. the method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method as claimed in claim 1, when it is characterized in that in supercritical carbon dioxide fluid, hatching, the time of stirring hatching is preferably 30 minutes.
6. the method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method as claimed in claim 1 is characterized in that cholesterol and the weight ratio of lecithin in the step 1) is preferably 1: 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008102005466A CN101683322B (en) | 2008-09-26 | 2008-09-26 | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008102005466A CN101683322B (en) | 2008-09-26 | 2008-09-26 | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101683322A CN101683322A (en) | 2010-03-31 |
| CN101683322B true CN101683322B (en) | 2012-07-18 |
Family
ID=42046857
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008102005466A Expired - Fee Related CN101683322B (en) | 2008-09-26 | 2008-09-26 | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101683322B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379847B (en) * | 2010-08-31 | 2013-03-13 | 华东理工大学 | Controlled synthesis method of nanometer polyenic taxusol liposome |
| CN101978952A (en) * | 2010-10-22 | 2011-02-23 | 广东医学院 | Method for preparing berberine hydrochloride liposome preparation |
| CN102475682B (en) * | 2010-11-30 | 2013-03-13 | 沈阳药科大学 | Berberine liposome and preparation method thereof |
| CN102488656A (en) * | 2011-02-23 | 2012-06-13 | 大连大学 | Method for preparing water-soluble drug liposome by utilizing reverse evaporating process of supercritical fluid |
| CN103449533B (en) * | 2012-05-29 | 2015-02-18 | 华东理工大学 | Supercritical carbon dioxide method for extraction separation of magnetosomes from magnetotactic bacteria |
| CN110448661B (en) * | 2019-08-13 | 2021-12-28 | 合肥市未来药物开发有限公司 | Zedoary turmeric oil mixed micelle and preparation method thereof |
| CN111001000B (en) * | 2019-12-10 | 2022-02-01 | 金陵科技学院 | Berberine hydrochloride FeAg nano-alloy liposome with light-controlled drug release and magnetic-controlled drug release and preparation method thereof |
| CN113143847B (en) * | 2021-03-09 | 2022-11-01 | 厦门大学 | Preparation method and application of long-acting low-toxicity insect-expelling ointment for skin and supercritical carbon dioxide fluid |
-
2008
- 2008-09-26 CN CN2008102005466A patent/CN101683322B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 严宾等.《超临界CO2法制备头孢唑啉钠脂质体》.《物理化学学报》.2006,第22卷(第02期),第226－229页. * |
| 平其能.《脂质体的制备》.《药剂学实验与指导》.中国医药科技出版社,1994,第194-195页. * |
| 裘丹等.《两种盐酸小檗碱脂质体制备方法的比较》.《中国化学会第十一届胶体与界面化学会议论文摘要集》.2007,第292－293页. * |
| 陈俊英等.《无水乙醇环境下超临界CO2萃取薯蓣皂素的研究》.《化学世界》.2007,(第06期),第349-356页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101683322A (en) | 2010-03-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101683322B (en) | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method | |
| Wang et al. | Cancer nanomedicines stabilized by π-π stacking between heterodimeric prodrugs enable exceptionally high drug loading capacity and safer delivery of drug combinations | |
| Wan et al. | Structure‐guided engineering of cytotoxic cabazitaxel for an adaptive nanoparticle formulation: Enhancing the drug safety and therapeutic efficacy | |
| Xing et al. | Studies on the oridonin-loaded poly (D, L-lactic acid) nanoparticles in vitro and in vivo | |
| Yi et al. | Cytotoxic effect of novel Flammulina velutipes sterols and its oral bioavailability via mixed micellar nanoformulation | |
| CN102225205B (en) | Tripterine nano structure lipid carrier and preparation method and application thereof | |
| US9283190B2 (en) | Highly efficient and long-acting slow-release formulation of poorly soluble drugs and preparation method thereof | |
| WO2008052410A1 (en) | A supermolecule composition of water-soluble coenzyme q10 and preparation method thereof | |
| CN107213467A (en) | A kind of preparation method of phospholipid complexes of curcumin | |
| CN104069071B (en) | A kind of silibinin sustained-release micropill of double-layer coatings and preparation method thereof | |
| CN102357075A (en) | Docetaxel nano preparation and preparation method thereof | |
| Shan et al. | Preparation of Icaritin-loaded mPEG-PLA micelles and evaluation on ischemic brain injury | |
| CN107049944B (en) | Polymer micelle capable of realizing simultaneous administration of sorafenib and curcumin and preparation method thereof | |
| CN114010799B (en) | Targeting degradable nano-drug carrier for photo-thermal synergistic chemotherapy and preparation method thereof | |
| CN106361724B (en) | A sustained release nanometer microsphere composition of 20(R) -ginsenoside Rg3 and its preparation method | |
| CN102961368A (en) | Curcumin nanosuspension and preparation method thereof | |
| Zheng et al. | Inositol hexanicotinate self-micelle solid dispersion is an efficient drug delivery system in the mouse model of non-alcoholic fatty liver disease | |
| CN103800286B (en) | A kind of 20 (S)-protopanoxadiol self-emulsion compositions and its preparation method and application | |
| CN105012234B (en) | A kind of 1,7-bis(3,4-dimethoxyphenyl)-1,6-heptadiene-3,5-dione polymer micelle and preparation method thereof and medical usage | |
| CN100423719C (en) | Nano granules of Docetaxel and preparing method | |
| CN103768022B (en) | A kind of self-assembled nanometer grain, Preparation Method And The Use that paclitaxel is conveyed as targeting | |
| CN103656669A (en) | Compound micelle-based nano-vector, and preparation method and application thereof | |
| CN1422618A (en) | 9-nitrocamptothecin solid dispersant and preparation method | |
| EP2845599B1 (en) | Chinese herbal compound anti-tumour nanopreparation and use thereof | |
| CN102793678B (en) | A kind of preparation method of the Docetaxel injection without tween |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120718 Termination date: 20180926 |