CN108030929A - A kind of preparation method of Lazer's property liposome comprising berberine - Google Patents
A kind of preparation method of Lazer's property liposome comprising berberine Download PDFInfo
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- CN108030929A CN108030929A CN201711375734.8A CN201711375734A CN108030929A CN 108030929 A CN108030929 A CN 108030929A CN 201711375734 A CN201711375734 A CN 201711375734A CN 108030929 A CN108030929 A CN 108030929A
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- liposome
- lazer
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- berberine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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- Pharmacology & Pharmacy (AREA)
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- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The present invention relates to a kind of preparation method of Lazer's property liposome comprising berberine, which is mainly made of phosphatidyl choline, polyglycol derivatization phospholipid, unsaturated phosphatide, berberine with poly- (2 ethylacrylic acid).The average grain diameter of the novel lipide is 435nm, and pH sensitive ranges are 5.2~7.0, and thermo-responsive scope is 39 DEG C~48 DEG C, has pH sensitiveness and heat sensitivity.In practical application, the Lazer's property liposome of the invention prepared, has the advantages that stability is good, and targeting is strong；Lazer's property method for preparing lipidosome of the present invention is simple and practicable at the same time.
Description
Technical field
The invention belongs to drug encapsulation technical field, more particularly to a kind of preparation of Lazer's property liposome comprising berberine
Method.
Background technology
Drug release material be it is a kind of there is the material of Drug controlled release, can by film through control system,
The mode Drug controlled release such as body diffusion system.Liposome is as one kind in numerous drug release materials, due to its preparation
Simply, harmless, non-immunogenicity reaction, easy the advantages that preparing, are paid close attention to by people.
Liposome is to be dispersed in water by phosphatide or other amphipathic molecules and formed, and has targeting, sustained release, biology
The features such as compatibility, be the good targeting vector of medicine.Due to liposome have the function of the characteristic of biomembrane and, mainly as cancer
Disease drug carrier is studied, it can transport directly to the active material of encapsulating on target cell, thus have " biological missile " it
Claim.But since traditional liposomal prepared by natural phospholipid, cholesterol is because stability is poor, product packaging rate is relatively low and targeting point
The reasons such as cloth is not good enough, using being above restricted, therefore a kind of novel lipide material with superperformance of exploitation seems, pole has must
Will.
In recent years, people have developed novel intelligent liposome to improve the stability of liposome, design and develop temperature-sensitive
The novel intelligent liposomes such as the quick liposome of liposome, pH, immunoliposome, magnetic liposome, photosensitive liposomes.Wherein most closed
Note is the quick liposomes of pH and thermal sensitive liposome, but they have the shortcomings that it is respective, such as the quick liposomes of pH in human body stabilization
Property is poor, and the toxic side effect of thermal sensitive liposome is larger.The pole therefore it provides a kind of more perfect method for preparing lipidosome seems
It is necessary.
For the present invention using having acid-sensitive poly- (2- ethylacrylic acids) modification blank liposome, the obtained quick liposomes of pH are quick
It is perceptual good；By selecting different lecithin materials and design proportioning that thermal sensitive liposome is made, its sensitiveness is good and poison is secondary makees
With smaller.Two kinds of liposomes are merged, are made while two kinds of liposome each shortcomings are improved by the present invention with blending method
There is Lazer's property liposome of two kinds of liposome good characteristics at the same time.
The content of the invention
In view of the deficiencies of the prior art, it is an object of the present invention to provide a kind of system of Lazer's property liposome comprising berberine
Preparation Method.
To achieve the above object, the present invention uses following scheme：A kind of preparation of Lazer's property liposome comprising berberine
Method, comprise the following steps：
(1) 5.5mol egg yolk lecithins and 4.5mol cholesterol are codissolved in 10mL chloroforms, 40 DEG C of rotary evaporation film forming,
24h is dried in vacuo, is added in 10mL PBS buffer, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains blank
Liposome solutions.
(2) with 1:6 mass ratio takes acid-sensitive dose poly- (2- ethylacrylic acids) to be reacted with tetradecylamine, and obtained 100mg gathers
(2- ethylacrylic acids) fat amide derivant, is added in the blank liposomes liquid solution of step 1 preparation, and 40 DEG C of stirring 24h, are stirred
It is not embedded into the derivative of liposome after the completion of mixing by the elution of SePharose CL-6B chromatographic columns using HBS buffer solutions as eluent,
Obtain the quick liposome solutions of pH.
(3) 0.9g dipalmitoylphosphatidylcholine (DPPC), 0.06g hydrogenated soya phosphatides (HSPC) and 0.04g bis- are weighed
Stearoyl monoethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), is codissolved in 10mL chloroforms, 60 DEG C of rotary evaporation film forming, very
The dry 24h of sky, is added in 10mLPBS buffer solutions, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains temperature-sensitive lipid
Liquid solution.
(4) the thermal sensitive liposome solution of the quick liposome solutions of the pH of 1 volume, 1 volume is added to chloroform/first of 4 volumes
Alcohol (volume ratio 2:1) mixed solution, rotary evaporation film forming, is dried in vacuo 24h, the berberine concentration for being added to 2 volumes is 1mg/
In the PBS buffer of the berberine of mL, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtain containing the double of berberine
Quick property liposome.
(5) Lazer's property liposome made from step (4) by the makrolon membrane filtration in 0.45 μm of aperture, is obtained into Lazer
Property liposome.
The beneficial effects of the present invention are：
(1) the selected thermal sensitive liposome raw material of the present invention are dipalmitoylphosphatidylcholine (DPPC), hydrogenated soybean
Phosphatide (HSPC), distearyl monoethanolamine-polyethylene glycol (DSPE-PEG2000), compared with traditional thermal liposome raw material,
Eliminate the lysophosphatide of ancillary drug release.Reason is：On the one hand, lysophosphatide can break the red blood cell in organism
Split, the amount of addition needs stringent control, and there are security risk；On the other hand, lysophosphatide is expensive, removes lysophosphatide
After can reduce liposome preparation cost.Due to eliminating the supplement of this raw material of lysophosphatide, invention increases two palm fibres
The dosage of palmitic acid phosphatidyl choline (DPPC), 90 are designed as by the weight ratio for the raw material for preparing thermal sensitive liposome:6:4.Be by
In the dipalmitoylphosphatidylcholine (DPPC) added phase transition temperature be 41 DEG C, closer to thermal sensitive liposome in human body
Sensitive temperature when working, drug release effect of the thermal sensitive liposome in human body can be lifted.Also achieve and the quick fat of pH at the same time
Plastid efficiently merges.
(2) present invention prepares the quick liposomes of pH by macromolecule modified method, i.e., using poly- (2- ethylacrylic acids) and ten
This high-molecular compound of (2- ethylacrylic acids) fat amide derivant poly- made from tetraalkyl amine modifies blank liposome.Make
With this macromolecule modified liposome with acid-sensitive, can not only make liposome that there is pH sensitiveness, meanwhile, alkalescence or in
In the quick working ranges of non-pH of property, which is rendered as extended configuration on the surface of liposome membrane, the quick liposomes of " cover " pH
Class eucaryotic cell structure, avoid the premature immune system by human body of the quick liposomes of pH and remove, extend the quick liposomes of pH and exist
Circulation time in organism.
Brief description of the drawings
Fig. 1 Lazers property liposome pH sensitive range test charts；
Means of differential scanning calorimetry (DSC) figure of Fig. 2 Lazers property liposome；
The grain size distribution of Fig. 3 Lazers property liposome.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with specific embodiment.
A kind of preparation of Lazer's property liposome comprising berberine of embodiment 1
(1) 5.5mol egg yolk lecithins and 4.5mol cholesterol are codissolved in 10mL chloroforms, 40 DEG C of rotary evaporation film forming,
24h is dried in vacuo, is added in 10mLPBS buffer solutions, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains blank fat
Plastid solution.
(2) with 1:6 mass ratio takes acid-sensitive dose poly- (2- ethylacrylic acids) to be reacted with tetradecylamine, and obtained 100mg gathers
(2- ethylacrylic acids) fat amide derivant, is added in the blank liposomes liquid solution of step 1 preparation, and 40 DEG C of stirring 24h, are stirred
It is not embedded into the derivative of liposome after the completion of mixing by the elution of SePharose CL-6B chromatographic columns using HBS buffer solutions as eluent,
Obtain the quick liposome solutions of pH.
(3) 0.9g dipalmitoylphosphatidylcholine (DPPC), 0.06g hydrogenated soya phosphatides (HSPC) and 0.04g bis- are weighed
Stearoyl monoethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), is codissolved in 10mL chloroforms, 60 DEG C of rotary evaporation film forming, very
The dry 24h of sky, is added in 10mL PBS buffer, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains temperature-sensitive fat
Plastid solution.
(4) the thermal sensitive liposome solution of the quick liposome solutions of the pH of 1 volume, 1 volume is added to chloroform/first of 4 volumes
Alcohol (volume ratio 2:1) mixed solution, rotary evaporation film forming, is dried in vacuo 24h, the berberine concentration for being added to 2 volumes is 1mg/
In the PBS buffer of the berberine of mL, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtain containing the double of berberine
Quick property liposome.
(5) Lazer's property liposome made from step (4) by the makrolon membrane filtration in 0.45 μm of aperture, is obtained into lipid
Body.
The pH sensitive ranges measure of 2 Lazer's property liposome of embodiment
Using the pH sensitive ranges of the Lazer's property liposome prepared in the determined by ultraviolet spectrophotometry embodiment of the present invention 1.
Each group Lazer property liposome is placed in 3.5~7.5 pH environment, the absorbance of different groups of solution is measured after a period of time,
The quick scopes of pH of Lazer's property liposome are measured 5.2~7.0, as shown in Figure 1.
The thermally sensitive temperature scope measure of 3 Lazer's property liposome of embodiment
The phase alternating temperature of the Lazer's property liposome prepared in the embodiment of the present invention 1 is measured using differential scanning calorimetry (DSC)
Degree.Lazer's property liposome is diluted to 20mg/mL, draws 10 μ L in aluminium dish, 20 DEG C~60 DEG C range scans, heating rate 2
DEG C/min, 39 DEG C~48 DEG C of phase transition temperature is measured, as shown in Figure 2.
The particle size determination of 4 Lazer's property liposome of embodiment
The particle diameter distribution of the Lazer's property liposome prepared in the embodiment of the present invention 1 is measured using laser fineness gage.Will
Lazer's property liposome is diluted to 20mg/mL, draws 20 μ L in cuvette, is measured after distilled water dilution with Laser Scattering Particle instrument
Particle diameter distribution, it is 435nm to measure Lazer's property liposome average grain diameter, as shown in Figure 3.
The drug release effect of 5 Lazer's property liposome of embodiment
The Lazer's property liposome prepared in above-described embodiment 1 shows through the quick scopes of pH and thermally sensitive temperature scope measure, in pH
Condition can accelerate the release of medicine in 5.2~7.0 and temperature in the range of 39 DEG C~48 DEG C, illustrate the liposome in biology
There is good controlled-release effect in vivo, improve the bioavailability of medicine.
The principle that embodiment described above is intended to be merely illustrative of the present uses preferable embodiment, but simultaneously
The limitation to the scope of the claims of the present invention cannot be considered as.It should be pointed out that for those of ordinary skill in the art, not
In the case of departing from present inventive concept, some deformations can also be made in some details and are changed, are regarded as institute of the present invention
The protection domain covered.
Claims (1)
1. a kind of preparation method of Lazer's property liposome comprising berberine, it is characterised in that comprise the following steps：
(1) 5.5mol egg yolk lecithins and 4.5mol cholesterol are codissolved in 10mL chloroforms, 40 DEG C of rotary evaporation film forming, vacuum
Dry 24h, is added in 10mLPBS buffer solutions, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains blank liposome
Solution.
(2) with 1:6 mass ratio takes acid-sensitive dose poly- (2- ethylacrylic acids) to be reacted with tetradecylamine, and poly- (the 2- second of 100mg is made
Base acrylic acid) fat amide derivant, it is added in the blank liposomes liquid solution of step 1 preparation, 40 DEG C of stirring 24h, have been stirred
It is not embedded into the derivative of liposome by the elution of SePharose CL-6B chromatographic columns using HBS buffer solutions as eluent after, obtains
The quick liposome solutions of pH.
(3) 0.9g dipalmitoylphosphatidylcholine (DPPC), 0.06g hydrogenated soya phosphatides (HSPC) and 0.04g distearyls are weighed
Acyl monoethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), is codissolved in 10mL chloroforms, and 60 DEG C of rotary evaporation film forming, vacuum is done
Dry 24h, is added in 10mL PBS buffer, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtains thermal sensitive liposome
Solution.
(4) the thermal sensitive liposome solution of the quick liposome solutions of the pH of 1 volume, 1 volume is added to the chloroform/methanol (body of 4 volumes
Product ratio 2:1) mixed solution, rotary evaporation film forming, is dried in vacuo 24h, and the berberine concentration for being added to 2 volumes is the Huang of 1mg/mL
In the PBS buffer of Lian Su, using Ultrasound Instrument every 2s~3s interval ultrasound 2min, obtain containing Lazer's property fat of berberine
Plastid.
(5) Lazer's property liposome made from step (4) by the makrolon membrane filtration in 0.45 μm of aperture, is obtained into Lazer's property fat
Plastid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201711375734.8A CN108030929B (en) | 2017-12-19 | 2017-12-19 | Preparation method of double-sensitive liposome containing berberine |
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| CN201711375734.8A CN108030929B (en) | 2017-12-19 | 2017-12-19 | Preparation method of double-sensitive liposome containing berberine |
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| CN108030929A true CN108030929A (en) | 2018-05-15 |
| CN108030929B CN108030929B (en) | 2021-02-09 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102210870A (en) * | 2011-06-09 | 2011-10-12 | 南京中医药大学 | Liposome composite phospholipid capable of adjusting phase-transition temperature and application thereof |
-
2017
- 2017-12-19 CN CN201711375734.8A patent/CN108030929B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102210870A (en) * | 2011-06-09 | 2011-10-12 | 南京中医药大学 | Liposome composite phospholipid capable of adjusting phase-transition temperature and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| WENTING ZHOU ET AL.: ""Characteristics, phase behavior and control release for copolymer–liposome with both pH and temperature sensitivities"", 《COLLOIDS AND SURFACES A: PHYSICOCHEMICAL AND ENGINEERING ASPECTS》 * |
| 王汝涛等: ""聚（2-乙基丙烯酸）脂肪酰胺衍生物构建的酸敏脂质体"", 《药学学报》 * |
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