CN1186011C - automatic liposome preparation apparatus and method for prepaving liplsome by same - Google Patents
automatic liposome preparation apparatus and method for prepaving liplsome by same Download PDFInfo
- Publication number
- CN1186011C CN1186011C CNB021394296A CN02139429A CN1186011C CN 1186011 C CN1186011 C CN 1186011C CN B021394296 A CNB021394296 A CN B021394296A CN 02139429 A CN02139429 A CN 02139429A CN 1186011 C CN1186011 C CN 1186011C
- Authority
- CN
- China
- Prior art keywords
- liposome
- pressure pump
- medicine
- organic
- phospholipid
- 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
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to an automatic liposome preparation device and a method for preparing liposome by the device. The preparation device comprises a high-pressure pump, a solution mixer, liposome particle diameter control equipment and a high-pressure ultrafiltration dialysis column and uses a principle of preparing the liposome by a solvent dispersion method. Phospholipid and liposoluble medicine dissolved in organic phases are rapidly mixed with a water solution at high pressure in the solution mixer by the high-pressure pump to form multiphase and multilayer liposome; the generated multiphase and multilayer liposome is forcibly and directly passes through a liposome squeezer to obtain large monochamber liposome at high pressure; medicine and organic solvent which are not coated in the liposome can be removed by the high-pressure ultrafiltration dialysis column. The present invention connects requisite devices of large-scale liposome production into a whole, integrates the whole process from material feed to liposome finished product acquisition, reduces the unnecessary intermediate processes, is capable of regulating the production scale according to requirements, has the controllable product quality and is not only suitable for liposoluble medicine but also suitable for water-soluble medicine.
Description
One, technical field
The invention belongs to field of medicaments, relate to the liposome preparation device, particularly this device of automatization's liposome preparation device and use prepares the method for liposome.
Two, background technology
Liposome (liposome is referred to as the fat bead again) is the hollow bead that is made of the phospholipid bilayer film.The main component that constitutes liposome is phospholipid and other lipids.Just find when phospholipid molecule is dispersed in the water multilamellar vesicle of similar biofilm structure that can nature formation ordered arrangement as far back as Britain scholar Bangham in 1961 and Standish.Each lipid layer of vesicle all constitutes with the phospholipid bimolecular of ordered arrangement; Separated by water between vesicle central authorities and each layer.People were commonly referred to as liposome with this hollow bead with similar biofilm structure of duplicature form afterwards.The character of liposome and particle diameter be according to its preparation method, the difference of material therefor and composition and widely different.Common by the double-deck immobilized artificial membrane number of plies that its structure comprised, charge property, function, purposes wait is classified to liposome.These classification method are because starting point difference has often been emphasized liposome characteristic in a certain respect, and various classification have different separately pluses and minuses.
(1) presses the shape and the particle size classification of liposome
Liposome can be by its shape and particle size classification.Different preparation methoies can form shape and the widely different liposome of particle diameter.That liposome can prepare is very little (approximately 25nm, the minimum grain size that theory can reach), also can be very big (1 μ m to 5 μ m is equivalent to the particle diameter of zooblast).They can comprise multilamellar phospholipid bilayer film, also can only contain monolayer phospholipid bilayer film.Because it is very big that the particle diameter of liposome is used influence to it, so be present the most frequently used sorting technique by its shape and particle size classification.
1, multilamelar liposome (claiming multilamellar liposome again, Multilamellar vesicles MLVs): the liposome that contains multilamellar phospholipid bilayer film is called multilamelar liposome, its particle diameter≤10 μ m (mostly being 300-5000nm).In general multilamelar liposome contains the phospholipid bilayer film more than five layers.The liposome that contains three to five layers of phospholipid bilayer film is called multiple layer liposome (Oligo-lamellar vesicles).For convenience, many times people completely call multilamelar liposome to multiple layer and multilamelar liposome.
2, small unilamellar vesicle (Small unilamellar vesicles SUVs): the liposome that is constituted by one deck phospholipid bilayer film.The particle diameter of small unilamellar vesicle is less than 50nm.Along with the particle diameter of liposome successively decreases, its spheroid radian increases, and causes the asymmetric of adventitia in the phospholipid bilayer film thereupon.It is especially outstanding that this inside and outside membrane assymmetry shows in small unilamellar vesicle.Taken 70% phospholipid such as its outer membrane of 25nm small unilamellar vesicle (thickness of phospholipid bilayer film is 4-5nm).Inside and outside membrane assymmetry makes the sphere of small unilamellar vesicle that very big tension force be arranged.So with respect to large unilamellar vesicle, the small unilamellar vesicle instability is easy to merge.
3, large unilamellar vesicle (Large unilamellar vesicles LUVs): the liposome that is constituted by one deck phospholipid bilayer film.The difference of itself and small unilamellar vesicle only is varying in size of particle diameter.The particle diameter of large unilamellar vesicle is generally about 80-200nm.Its long time in blood is imitated, high medicine parcel amount, and stability has determined that large unilamellar vesicle is ideal intravenously administrable pharmaceutical carrier.The liposome medicament majority that has gone on the market at present is a large unilamellar vesicle.
(2) press the surface charge property classification
1, neutral fat plastid: mainly by lecithin, the liposome that uncharged phospholipid and other uncharged lipids constitute.
2, elecrtonegativity liposome: with regard to liposome generally speaking, be elecrtonegativity.Contain the electronegative phospholipid of part in its phospholipid component, as phosphatidyl silk amino acid (PS), phosphatidylinositols (PI), and phosphatidyl glycerol fat (PG).
3, electropositive liposome: liposome is electropositive.Contain the positively charged chemical compound of part in its phospholipid component, as two-octadecane-two-methyl amine salt (DODAC), 18 ammonia.The electropositive liposome has been subjected to paying close attention to widely as the carrier of non-viral gene medicine in recent years.
(3) by functional classification
1. conventional liposome: comprise above-mentioned unilamelar liposome, multilamelar liposome and multiphasic liposomes etc.
2. long-acting liposome: can in blood, keep longer a period of time.Its surface is many with Polyethylene Glycol, Jia Ji Ju oxazoline, and the polyethylene pyrrole is modified.
3. intelligent liposome: utilize the inside and outside special physicochemical property of some health and the liposome of constructing.This lipoid plastid changes with the variation of external condition.Can be regularly, fixed point, quantitative release medicine.Comprised temperature-sensitive, acid-sensitive, photosensitive, magnetosensitive, controllable release and target administration.
(4) press liposome development division of history into periods classification
This classification method mainly lays particular emphasis on liposome has breakthrough liposome function to improve index as the developing history of drug administration carrier and important generation, is difficult to a generally acknowledged unified saying.
1, first generation liposome: medicine is by natural or synthetic phospholipid, and it is the most basic that the duplicature that cholesterol is formed wraps up, the simplest liposome structure.Major function is for reducing toxicity and increasing drug effect.The liposome medicament produced of having gone on the market belongs to this type of more, and (trade name, AmBisome), liposomal doxorubicin (Myocet) and liposome daunorubicin (DaunoXome) be by the FDA authentication, listing production such as: liposome amphotericin.
2, second filial generation liposome: the liposome that the surface is modified with Polyethylene Glycol-phospholipid derivative (PEG-lipids).Liposome after the modification has improved the holding time of medicine in blood, has increased the passive target function of medicine.Polyethylene Glycol-liposomal doxorubicin (Doxil) is by FDA authentication, and listing is produced, and multiple Polyethylene Glycol-liposome medicament has also entered the clinical II/III phase and tested.
3, third generation liposome: the surface is used Polyethylene Glycol-phospholipid derivative (PEG-lipids) modification and liposome introduce the targeting group in the Polyethylene Glycol terminal.Improved initiatively target function, thus the drug effect of significantly improving.These targeting groups can be monoclonal antibody, polysaccharide, polypeptide and vitamin.This lipoid plastid medicine has also entered the clinical I/II phase and has tested.
4, the 4th generation liposome: make liposome have targeting by modification, controllable sustained-release, acid-sensitive amalgamation and other controllabilities, thereby the drug effect of significantly improving.This lipoid plastid medicine has certain controllability, also is called intelligent liposome.The 4th generation liposome also be in the laboratory research stage.
(5) by purposes and route of administration classification
1, intravenous drip liposome.
2, external (percutaneous dosing) liposome.
3, oral liposome.
4, medicament for the eyes liposome.
5, respiratory tract administration liposome.
6, mucosa delivery liposome.
7, intramuscular injection and topical liposome.
8, polypeptide, albumen and gene drug delivery liposome.
9, liposome bacterin.
10, the liposome of other purposes.
(6), traditional liposomal industrial production process and defective
1), mechanical dispersion method
Various phospholipid dressing (if fat-soluble medicine also should be included in this part) are dissolved in the organic solvent uniformly, make uniform thin film after organic solvent is steamed.In the phospholipid membrane container, add water or the buffer or the water solublity medicinal liquid of formula ratio, make thin film be scattered in water with mechanical dispersion method and both can form liposome.When liposome forms, get final product success water soluble drug is wrapped in the liposome empty pocket, or oil-soluble medicine is wrapped in the liposome duplicature.The particle diameter of liposome can be contracted to the grain size that needs with the different-grain diameter control device.The dialysis apparatus that can use of Bao Guo medicine is not separated.
The limitation of mechanical dispersion method is: phospholipid membrane forms influence and the restriction that step is subjected to solvent evaporation equipment, both has been difficult for making uniform phospholipid membrane, also is difficult for large-scale production.The medicine parcel efficient of this method depends on the quality of immobilized artificial membrane in aqueous phase mechanical dispersion degree.If immobilized artificial membrane mechanical dispersion degree in water is bad in addition, the heterogeneous multilamelar liposome particle diameter of formation is too big, can make the very difficulty of dwindling of liposome particle diameter.
2), solvent dispersion method
With with the miscible organic solvent dissolution phospholipid dressing of water and (or) fat-soluble medicine, the organic phospholipid drug solution is added slowly with in the aqueous solution that stirs, organic solvent disperses back phospholipid to be forced to form heterogeneous multilamelar liposome drug solution at aqueous phase.Influence of various factors such as the physicochemical property of the ratio that liposome particle diameter that forms and bag dose are subjected to organic solution and water, mixing speed, solution temperature, aqueous solution pH, solution particle intensity, medicine, the kind of phospholipid and prescription.In general, the content of organic solution amount in final liquid should be between 7%-20%.
Because the content of this preparation method requirement organic solution amount in final liquid should be between 7%-20%, and the dissolubility of phospholipid in solvent for use can not be too big, so the liposome concentration of this method preparation is very low, if there is not effective concentrator, the requirement that the liposome medicinal liquid concentration of making does not reach clinical application concentration.Simultaneously, the medicine parcel efficient of this method and the size of liposome particle diameter depend on the mixing speed of the rate of addition and the mixed solution of organic solution in the preparation process, the rate of addition of organic solution is difficult to control when large-scale production, and mechanical dispersion speed also difficulty reach desirable level.
3), surfactant-dispersed method
The aforementioned two kinds of methods of many macromolecular compounds are difficult to the medicine of parcel effective dose.So produced the surfactant-dispersed method thus.This method is dissolved in macromolecular compound, phospholipid, surfactant in the aqueous solution by a certain percentage, utilizes dialysis apparatus slowly surfactant to be removed and promptly can be made into liposome medicament.This method has been used to the liposome gene medicine, the preparation of liposome protein drug and liposome polypeptide drugs.
This preparation method requires the CMC value of surfactant between 5-20, and the surfactant of Shi Yonging can not influence macromolecular biological activity simultaneously.In order to improve the embedding rate of medicine, usually in prescription, use charged phospholipid or the macromolecule modified phospholipid of process.
Three, summary of the invention
Purpose of the present invention is intended to design a kind of automatization liposome preparation device and uses this device to prepare the method for liposome.This device has been used the principle that the solvent dispersion legal system is equipped with liposome, to be dissolved in the phospholipid of organic facies and fat-soluble medicine and aqueous solution by high-pressure pump and make it to form heterogeneous multilamellar liposome, and so just can avoid the rate of addition of conventional solvent dispersion method organic solution to be difficult to the defective that control and mechanical dispersion speed difficulty reach desirable level in solution mixer mesohigh rapid mixing.
To achieve these goals, the technical solution adopted in the present invention is, a kind ofly is used to prepare fat-soluble and automatization's liposome preparation device water soluble drug, is characterized in that this device comprises:
Monophosphatide NaOH solution tank NaOH 1, phospholipid solution jar 1 is communicated with first organic high-pressure pump 3 by first three-way valve 2;
One medicine liquid tank 5, medicine liquid tank 5 is communicated with second organic high-pressure pump 4;
First organic high-pressure pump 3 and second organic high-pressure pump 4 are communicated with a blender 6 respectively; The squeezer 7 that blender 6 and dwindles the liposome particle diameter links to each other, and communicates with an arrival end of a liposome storage tank 8; The port of export of liposome storage tank 8 bottoms by second three-way valve 9 with first three-way valve 2, be connected disc type control valve 10 and be connected respectively;
One buffering flow container 12, the arrival end of buffering flow container 12 is communicated with another arrival end of liposome jar 8, and is communicated with a control valve 13, and the other end of control valve 13 is connected with an end of the ultrafilter 15 that a purification liposome is used; The port of export of buffering flow container 12 is connected with a high-pressure pump 14 by the 3rd three-way valve 11; The other end of the high pressure ultrafiltration dialysis post of using by high-pressure pump 14 and purification liposome 15 is connected;
The high pressure ultrafiltration dialysis post 15 that the purification liposome is used also links to each other with a waste liquid tank 17 by another control valve 16.
Other characteristics of this device are, said first and second organic high-pressure pump 3,4 and high-pressure pump 14 are single cylinder, twin-tub piston type organic solution high-pressure pump, rotary-type organic solution high-pressure pump, reciprocating type organic solution high-pressure pump, the high performance liquid chromatogram infusion pump, preparation any of high performance liquid chromatogram infusion pump.
Said storage tank 8 is any in the fluid reservoir of water-bath temperature control fluid reservoir, steam temperature control fluid reservoir, automatically controlled temperature fluid reservoir, no temperature regulating device.
The said squeezer 7 that dwindles the liposome particle diameter is, band controllable temperature device or do not have the liposome membrane squeezer of temperature regulating device, or homogenizer, or microgranule fluidization processor or other liposome particle diameter control appliance.
All have temperature regulating device on said phospholipid solution jar 1, squeezer 7, liposome jar 8 and the buffering flow container 12.
The high pressure ultrafiltration dialysis post 15 that said purification liposome is used is Hollow Fiber Ultrafiltration dialysis cartridge, or gel ultrafiltration dialysis post, or the membrane ultrafiltration dialysis cartridge.
Adopting the method for preparing liposome of above-mentioned automatization liposome preparation device is that whole liposome production process is carried out under nitrogen protection, and carries out by the following method:
1) fat-soluble medicine of recipe quantity, lecithin, cholesterol, the oleic acid of liposome material and the vitamin E of antioxidant of liposome material are added organic solvent, heating makes it fully to be injected with in the phospholipid solution jar 1 after the dissolving; If during with this method coated water-soluble medicine, water soluble drug makes it fully dissolving back injection medicine liquid tank 5 at aqueous solution or buffer heating;
2) organic phospholipid medicine and aqueous solution are pressed in the blender 6 with first organic high-pressure pump 3 and second organic high-pressure pump 4 respectively, make that organic solvent is quick, be dispersed in the water at blender mesohigh machine phospholipid medicine and aqueous solution rapid mixing, force phospholipid to form heterogeneous multilamelar liposome drug solution;
3) can obtain the liposome solutions that particle diameter has dwindled by squeezer 7; Dwindling the technical process of particle diameter can carry out repeatedly, and the liposome solutions in the liposome storage tank 8 can be by organic high-pressure pump 3, blender 6 and squeezer 7 circulation repetitive operations, until the liposome solutions that reaches required particle size range;
4) then, liposome solutions is pressed into high pressure ultrafiltration dialysis post 15 through high-pressure pump 14, organic solvent and the medicine that is not wrapped can dialyse out; Carry out the liposome medicament product that the ultrafiltration dialysis process promptly can obtain purifying repeatedly, final products should be uniform milky solutions, do not precipitate lamination;
5) the above-mentioned product bottling back that obtains is preserved under 2 ℃ of-8 ℃ of conditions.
The method for preparing liposome of above-mentioned use automatization liposome preparation device; Described fat-soluble medicine is:
1. antibacterials: comprise daunorubicin, amphotericin B;
2. cancer therapy drug: comprise 5-fluorouracil, doxorubicin hydrochloride, Farmorubine Hydrochloride, vincristine sulfate, topotecan, mitoxantrone, paclitaxel, Docetaxel; And
3. fat-soluble medicine: comprise miconazole, itraconazole, econazole, ketoconazole, fluconazol, fenticonazole, sulconazole, tioconazole, Sertaconazole, terconazole (triaconazole), croconazole, isoconazole, fluorine triaconazole and water soluble drug;
Described liposome material lecithin is: Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline, synthetic lecithin, sphingomyelins.
The fat-soluble medicine consumption of described recipe quantity is 0.1%-16% (mol ratio); Consumption as the phospholipid of liposome material is 50%-80% (mol ratio); Consumption as the cholesterol of liposome material is 0-30% (mol ratio); Oleic consumption as the liposome material is 0-10% (mol ratio); Consumption as the vitamin E of antioxidant is 0-0.4% (mol ratio).
The particle diameter of described liposome can be that the particle diameter of liposome is 80nm～240nm.
Device of the present invention constitutes jointly interlock with organic solution high-pressure pump, solution mixer, liposome particle diameter control appliance and high pressure ultrafiltration dialysis post and becomes one, breathe out from being dosed into whole process one gas that obtains the liposome finished product, reduced unnecessary pilot process, production scale can be regulated according to need, product quality controllable, be not only applicable to fat-soluble, also be applicable to water soluble drug.Experimental results show that particle diameter is even, narrow diameter distribution with the controlled amount of the liposome of this device preparation.Pro-liposome with this device preparation can form effective pH or ion gradient, thereby can utilize the high efficiency parcel weak-alkaline and weak-acidic of gradient method medicine.
Four, description of drawings
Fig. 1 is an automatization of the present invention liposome preparation device sketch map;
Fig. 2 is the frozen section electron micrograph of the liposome made of automatization of the present invention liposome preparation device.
Five, the specific embodiment
For a more clear understanding of the present invention, the present invention is described in further detail for the embodiment that finishes according to technical scheme of the present invention that provides below in conjunction with the inventor.
5.1, apparatus and the liposome technological process of production
5.1.1, apparatus figure
Referring to Fig. 1, device comprises:
Monophosphatide NaOH solution tank NaOH 1, phospholipid solution jar 1 is communicated with first organic high-pressure pump 3 by first three-way valve 2;
One medicine liquid tank 5, medicine liquid tank 5 is communicated with second organic high-pressure pump 4;
First organic high-pressure pump 3 and second organic high-pressure pump 4 are communicated with a blender 6 respectively; The squeezer 7 that blender 6 and dwindles the liposome particle diameter links to each other, and communicates with an arrival end of a liposome storage tank 8; The port of export of liposome storage tank 8 bottoms by second three-way valve 9 with first three-way valve 2, be connected disc type control valve 10 and be connected respectively;
One buffering flow container 12, the arrival end of buffering flow container 12 is communicated with another arrival end of liposome jar 8, and is communicated with a control valve 13, and the other end of control valve 13 is connected with an end of the ultrafilter 15 that a purification liposome is used; The port of export of buffering flow container 12 is connected with a high-pressure pump 14 by the 3rd three-way valve 11; The other end of the high pressure ultrafiltration dialysis post of using by high-pressure pump 14 and purification liposome 15 is connected;
The high pressure ultrafiltration dialysis post 15 that the purification liposome is used also links to each other with a waste liquid tank 17 by another control valve 16.
First and second organic high-pressure pump 3,4 and high-pressure pump 14 are: single cylinder, twin-tub piston type organic solution high-pressure pump, rotary-type organic solution high-pressure pump, reciprocating type organic solution high-pressure pump, high performance liquid chromatogram infusion pump, preparation any of high performance liquid chromatogram infusion pump.
The squeezer 7 that dwindles the liposome particle diameter is, band controllable temperature device or do not have the liposome membrane squeezer (extruder) of temperature regulating device, or homogenizer (homogenizer), or microgranule fluidization processor (microfluidizer processor) or other liposome particle diameter control appliance.
All have temperature regulating device on phospholipid solution jar 1, squeezer 7, liposome jar 8 and the buffering flow container 12.
The high pressure ultrafiltration dialysis post 15 that said purification liposome is used is Hollow Fiber Ultrafiltration dialysis cartridge, or gel ultrafiltration dialysis post, or the membrane ultrafiltration dialysis cartridge.
Above-mentioned liposome preparation Design of device is equipped with the principle of liposome based on the solvent dispersion legal system,, makes it fully, mixes effectively, uniformly and generate liposome respectively with organic solvent and aqueous solution high pressure, injecting mixer fast with two high pressure pumps; The plurality of step and the equipment of preparation liposome are reasonably linked together, make a concentrated step of liposome preparation, particle diameter control, purification and liposome finish, avoided the transfer repeatedly of preparation intermediate, reduced unnecessary pilot process; Production scale can be regulated according to need, product quality controllable; Install widely applicablely, can be used for preparing fat-soluble and water soluble drug.
5.1.2, production technology
Whole liposome production process is carried out under nitrogen protection.
Recipe quantity lecithin, cholesterol and fat-soluble medicine add organic solvent, and heating makes it to inject organic solution storage tank 1 after the fully dissolving.If during with this method coated water-soluble medicine, water soluble drug makes it fully dissolving back injection aqueous solution storage tank 5 at aqueous solution or buffer heating.Organic phospholipid medicine and aqueous solution are pressed in the blender 6 with high-pressure pump 3 and 4 respectively, make that organic solvent is quick, be dispersed in the water at blender mesohigh machine phospholipid medicine and aqueous solution rapid mixing, force phospholipid to form heterogeneous multilamelar liposome drug solution.Can obtain the liposome solutions that particle diameter has dwindled by particle diameter controller 7.Dwindling the technical process of particle diameter can carry out repeatedly, and the liposome solutions in the liposome storage tank 8 can pass through high-pressure pump 3, blender 6 and 7 these circulation repetitive operations of particle diameter controller, until the liposome solutions that reaches required particle size range.Then, liposome solutions is pressed into high pressure ultrafiltration dialysis post 15 through high-pressure pump 14, organic solvent and the medicine that is not wrapped can dialyse out.Carry out the liposome medicament product that the ultrafiltration dialysis process promptly can obtain purifying repeatedly.Final products should be uniform milky solutions, do not precipitate lamination.Should under 2 ℃ of-8 ℃ of conditions, preserve after the product bottling.
Below be the embodiment that the inventor provides, the present invention is not limited to these embodiment.
Embodiment 1: use automatization's liposome preparation device to prepare the liposome of 100nm particle diameter
The liposome prescription:
| Title | Molecular weight | Mole (%) | Inventory (g) |
| Lecithin | 760 | 60 | 18.24 |
| Cholesterol | 386.7 | 39 | 6.03 |
| Vitamin E | 472 | 1 | 0.19 |
With the lecithin of recipe quantity, cholesterol and vitamin E add 20ml ethanol, are heated to 45 ℃ of stirrings and make it to dissolve fully in the organic fluid reservoir of back adding, 180ml water for injection are added in the aqueous solution fluid reservoir the built-in double-deck 100nm filter membrane of squeezer.Start infusion pump, ethanol adjuvant solution and 180ml water for injection are prepared blender and the squeezer that pump A and B are pressed into novel automatic liposome preparation device by high performance liquid chromatography respectively.It is 20-30: 70-80 that the regulating gradient pump makes the flow-rate ratio of water pump and organic facies pump, collects the also particle diameter of testing product, liposome solutions can be pumped into blender and squeezer again as excessive.Finally can get the liposome solutions of particle size range at 80-180nm.Then, liposome solutions is pressed into high pressure ultrafiltration dialysis post 15 through high-pressure pump 14, organic solvent can be dialysed out by bag.Carry out the liposome medicament product that the ultrafiltration dialysis process promptly can obtain purifying repeatedly.
With the liposome that has prepared different-grain diameter with the quadrat method inventor.The frozen section electron micrograph of the thin film of preparation liposome institute different-grain diameter and the corresponding liposome of making as shown in Figure 2
Embodiment 2: use automatization of the present invention liposome preparation device to prepare the liposome that can be used for pH gradient method packaging medicine of 100nm particle diameter
With the liposome of device of the present invention preparation can be used for wrapping up useful traditional pH gradient method medicine that can wrap up, only verified with the example that is prepared as of liposomal doxorubicin.
The liposome prescription:
| Title | Molecular weight | Mole (%) | Inventory (g) |
| Lecithin | 760 | 60 | 18.24 |
| Cholesterol | 386.7 | 39 | 6.03 |
| Vitamin E | 472 | 1 | 0.19 |
Lecithin with recipe quantity, cholesterol and vitamin E add 20ml ethanol, are heated to 45 ℃ of stirrings and make it to dissolve fully in the organic fluid reservoir of back adding, with the 400mM of 180ml, the pH4 tartaric acid solution adds in the aqueous solution fluid reservoir, the built-in double-deck 100nm filter membrane of squeezer.Start infusion pump, with ethanol adjuvant solution and the 400mM tartaric acid solution prepare blender and the squeezer that pump A and B are pressed into novel automatic liposome preparation device by high performance liquid chromatography respectively.It is 20-30: 70-80 that the regulating gradient pump makes the flow-rate ratio of water pump and organic facies pump, collects the also particle diameter of testing product, liposome solutions can be pumped into blender and squeezer again as excessive.Finally can get the liposome solutions of particle diameter at 120nm ± 40nm.Then, liposome solutions is pressed into high pressure ultrafiltration dialysis post 15 through high-pressure pump 14, uses 400mM, the pH4 tartaric acid solution replenishes liquid as dialysis organic solvent is removed.Carry out the liposome that can be used for pH gradient method packaging medicine that the ultrafiltration dialysis process promptly can obtain purifying repeatedly.
The 50mg doxorubicin hydrochloride is added 2ml be surrounded by 400mM, the tartaric liposome solutions of pH4 treats that doxorubicin hydrochloride dissolves the Na of back with pH10 fully
2CO
3The soda acid of solution regulator solution is put the bottle 20min in 60 ℃ of water-baths that fills medicinal liquid to pH8.Ultracentrifugation detects proof medicine embedding rate and reaches more than 99%.
Embodiment 3: use novel automatic liposome preparation device to prepare liposoluble medicinal liposome
Apparatus of the present invention can be used for preparing all liposoluble medicinal liposomes, are only verified with the example that is prepared as of liposome miconazole.
The medicinal liposome prescription:
| Title | Molecular weight | Mole (%) | Inventory (g) |
| Lecithin | 760 | 87 | 19 |
| Cholesterol | 386.66 | 9.2 | 1.0 |
| Oleic acid | 479.15 | 15.0 | 2.5 |
| Miconazole nitrate | 282 | 15 | 0.3 |
| Azone | 281 | 0.1 | 0.01 |
| Vitamin E | 472 | 0.2 | 0.03 |
| Oleum menthae | 0.2 | 0.3 |
Recipe quantity lecithin, cholesterol, oleic acid, miconazole nitrate, azone, vitamin E and Oleum menthae add 40ml ethanol, be heated to 45 ℃ of stirrings and make it to dissolve fully in the organic fluid reservoir of back adding, in the pure adding aqueous solution fluid reservoir with 180ml, the built-in double-deck 200nm filter membrane of squeezer.Start infusion pump, with ethanol adjuvant solution and water prepare blender and the squeezer that pump A and B are pressed into novel automatic liposome preparation device by high performance liquid chromatography respectively.It is 20-30: 70-80 that the regulating gradient pump makes the flow-rate ratio of water pump and organic facies pump, collects the also particle diameter of testing product, liposome solutions can be pumped into blender and squeezer again as particle diameter is excessive.Finally can get the liposome medicament solution of particle diameter at 210nm+30nm.Because the liposome miconazole will be used for the treatment of dermatosis, so ethanol need not removed.Need not employ the ultrafiltration dialysis device in the production.
Get in the liposome miconazole solution that the 400ml pure water adds 200ml slowly.Adding the fashionable blunting of attention of answering stirs.Final products are uniform milky solution, do not precipitate lamination.Compare before diluting back liposome particle diameter 212nm ± 30nm and mixing and do not have significant change.Under 2 ℃-15 ℃ and lucifuge condition, preserve after the product fill.
Embodiment in sum can exemplify various, and the inventor proves in a large amount of experiments, as long as in the scope of the claim protection that the present invention proposes, can reach purpose of the present invention, does not exemplify one by one at this.
Claims (10)
1. one kind is used to prepare fat-soluble and automatization's liposome preparation device water soluble drug, it is characterized in that this device comprises:
Monophosphatide NaOH solution tank NaOH (1), phospholipid solution jar (1) is communicated with first organic high-pressure pump (3) by first three-way valve (2);
One medicine liquid tank (5), medicine liquid tank (5) is communicated with second organic high-pressure pump (4);
First organic high-pressure pump (3) and second organic high-pressure pump (4) are communicated with a blender (6) respectively; The squeezer (7) that blender (6) and dwindles the liposome particle diameter links to each other, and communicates with an arrival end of a liposome storage tank (8); The port of export of liposome storage tank (8) bottom by second three-way valve (9) with first three-way valve (2), be connected disc type control valve (10) and be connected respectively;
One buffering flow container (12), the arrival end of buffering flow container (12) is communicated with another arrival end of liposome jar (8), and is communicated with a control valve (13), and the other end of control valve (13) is connected with an end of the ultrafilter (15) that a purification liposome is used; The port of export of buffering flow container (12) is connected with a high-pressure pump (14) by the 3rd three-way valve (11); The other end of the high pressure ultrafiltration dialysis post of using by high-pressure pump (14) and purification liposome (15) is connected;
The high pressure ultrafiltration dialysis post (15) that the purification liposome is used also links to each other with a waste liquid tank (17) by another control valve (16).
2. according to automatization's liposome preparation device of claim described 1, it is characterized in that, said first and second organic high-pressure pump (3,4) and high-pressure pump (14) are single cylinder, twin-tub piston type organic solution high-pressure pump, rotary-type organic solution high-pressure pump, reciprocating type organic solution high-pressure pump, the high performance liquid chromatogram infusion pump, preparation any of high performance liquid chromatogram infusion pump.
3. according to automatization's liposome preparation device of claim described 1, it is characterized in that said storage tank (8) is any in the fluid reservoir of water-bath temperature control fluid reservoir, steam temperature control fluid reservoir, automatically controlled temperature fluid reservoir, no temperature regulating device.
4. according to automatization's liposome preparation device of claim described 1, it is characterized in that, the said squeezer (7) that dwindles the liposome particle diameter is band controllable temperature device or the liposome membrane squeezer that does not have temperature regulating device, or homogenizer, or microgranule fluidization processor or other liposome particle diameter control appliance.
5. according to automatization's liposome preparation device of claim described 1, it is characterized in that, all have temperature regulating device on said phospholipid solution jar (1), squeezer (7), liposome jar (8) and the buffering flow container (12).
6. according to automatization's liposome preparation device of claim described 1, it is characterized in that the high pressure ultrafiltration dialysis post (15) that said purification liposome is used is the Hollow Fiber Ultrafiltration dialysis cartridge, or gel ultrafiltration dialysis post, or the membrane ultrafiltration dialysis cartridge.
7. realize that the described automatization of claim 1 liposome preparation device prepares the method for liposome; It is characterized in that whole liposome production process is carried out, and carries out by the following method under nitrogen protection:
1) fat-soluble medicine of recipe quantity, lecithin, cholesterol, the oleic acid of liposome material and the vitamin E of antioxidant of liposome material are added organic solvent, heating makes it fully to be injected with in the phospholipid solution jar (1) after the dissolving; During with this method coated water-soluble medicine, water soluble drug makes it fully dissolving back injection medicine liquid tank (5) at aqueous solution or buffer heating;
2) use first organic high-pressure pump (3) and second organic high-pressure pump (4) to be pressed in the blender (6) respectively organic phospholipid medicine and aqueous solution, make that organic solvent is quick, be dispersed in the water at blender mesohigh machine phospholipid medicine and aqueous solution rapid mixing, force phospholipid to form heterogeneous multilamelar liposome drug solution;
3) promptly obtain the liposome solutions that particle diameter has dwindled by squeezer (7); Dwindle the technical process of particle diameter and carry out repeatedly, the liposome solutions in the liposome storage tank (8) is by organic high-pressure pump (3), blender (6) and squeezer (7) circulation repetitive operation, until the liposome solutions that reaches required particle size range;
4) then, liposome solutions is pressed into high pressure ultrafiltration dialysis post (15) through high-pressure pump (14), organic solvent and the medicine that is not wrapped can dialyse out; Carry out the ultrafiltration dialysis process repeatedly and promptly obtain the liposome medicament product of purifying, final products are uniform milky solutions, precipitation not, lamination;
5) the above-mentioned product bottling back that obtains is preserved under 2 ℃ of-8 ℃ of conditions.
8. the method for preparing liposome according to the described automatization of claim 7 liposome preparation device; It is characterized in that described fat-soluble medicine is:
1. antibacterials: comprise daunorubicin, amphotericin B;
2. cancer therapy drug: comprise 5-fluorouracil, doxorubicin hydrochloride, Farmorubine Hydrochloride, vincristine sulfate, topotecan, mitoxantrone, paclitaxel, Docetaxel; And
3. fat-soluble medicine: comprise miconazole, itraconazole, econazole, ketoconazole, fluconazol, fenticonazole, sulconazole, tioconazole, Sertaconazole, terconazole (triaconazole), croconazole, isoconazole, fluorine triaconazole and water soluble drug;
Described liposome material lecithin is Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline, synthetic lecithin, sphingomyelins.
9. the method for preparing liposome according to the described automatization of claim 7 liposome preparation device; It is characterized in that the fat-soluble medicine consumption of described recipe quantity is the 0.1-16% mol ratio; Consumption as the phospholipid of liposome material is the 50-80% mol ratio; Consumption as the cholesterol of liposome material is the 0-30% mol ratio; Oleic consumption as the liposome material is the 0-10% mol ratio; Consumption as the vitamin E of antioxidant is the 0-0.4% mol ratio.
10. the method for preparing liposome according to the described automatization of claim 7 liposome preparation device; It is characterized in that the particle diameter of described liposome is 80nm～240nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021394296A CN1186011C (en) | 2002-09-17 | 2002-09-17 | automatic liposome preparation apparatus and method for prepaving liplsome by same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021394296A CN1186011C (en) | 2002-09-17 | 2002-09-17 | automatic liposome preparation apparatus and method for prepaving liplsome by same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1483400A CN1483400A (en) | 2004-03-24 |
| CN1186011C true CN1186011C (en) | 2005-01-26 |
Family
ID=34147444
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021394296A Expired - Fee Related CN1186011C (en) | 2002-09-17 | 2002-09-17 | automatic liposome preparation apparatus and method for prepaving liplsome by same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1186011C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109937046A (en) * | 2016-09-09 | 2019-06-25 | 艾利西斯股份有限公司 | Liposome anti-cancer composition |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101617990B (en) * | 2009-07-31 | 2013-01-23 | 中国人民解放军第三军医大学 | Liposome preparation device and method for preparing liposome by using same |
| CN101912363A (en) * | 2010-07-29 | 2010-12-15 | 蔡海德 | Dissolving ultrafiltration-spray drying-molecule dispersion coating-hydration palletizing-freeze drying method for preparing liposome combination medicine |
| CN105012152B (en) * | 2014-04-23 | 2018-01-02 | 胡权 | The preparation facilities and preparation method of a kind of liposome |
| CN105434504B (en) * | 2014-08-29 | 2021-10-15 | 黑龙江科伦制药有限公司 | Fat emulsion tail liquid treatment device and method |
| CN113397978A (en) * | 2021-07-16 | 2021-09-17 | 江西科技师范大学 | System for preparing and purifying nanoliposome |
| CN115674769B (en) * | 2022-11-21 | 2023-03-21 | 苏州艾特森制药设备有限公司 | Intelligent preparation control method and equipment for liposome extrusion |
-
2002
- 2002-09-17 CN CNB021394296A patent/CN1186011C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109937046A (en) * | 2016-09-09 | 2019-06-25 | 艾利西斯股份有限公司 | Liposome anti-cancer composition |
| JP2019533006A (en) * | 2016-09-09 | 2019-11-14 | アイリシス・インコーポレイテッド | Liposome anticancer composition |
| EP3509605A4 (en) * | 2016-09-09 | 2020-05-27 | Irisys, Inc. | Liposomal anticancer compositions |
| US11033520B2 (en) | 2016-09-09 | 2021-06-15 | Irisys, Inc. | Liposomal anticancer compositions |
| AU2017324718B2 (en) * | 2016-09-09 | 2022-09-29 | Irisys, Inc. | Lipsomal anticancer compositions |
| JP2022172133A (en) * | 2016-09-09 | 2022-11-15 | アイリシス・インコーポレイテッド | Liposomal anticancer compositions |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1483400A (en) | 2004-03-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| FI104464B (en) | A process for the preparation of heterovesicular liposomes | |
| Gulati et al. | Parenteral drug delivery: a review | |
| JP3676976B2 (en) | Control of the amount of drug encapsulated in multivesicular liposomes | |
| CN101991538B (en) | TPGS-containing liposome composition and application thereof | |
| CN101244039B (en) | Novel method for preparing indissoluble medicaments liposome preparations | |
| CN102188377B (en) | Method for preparing medicine encapsulating liposome | |
| CN1186011C (en) | automatic liposome preparation apparatus and method for prepaving liplsome by same | |
| CA2505520A1 (en) | Protein-stabilized liposomal formulations of pharmaceutical agents | |
| JP2008231086A (en) | Emulsion composition containing prostaglandin e1 | |
| US20140161876A1 (en) | Liposome-containing preparation utilizing dissolution aid, and method for producing same | |
| US20230255894A1 (en) | Method of preparing lipid vesicles | |
| Liu et al. | Lecithin/isopropyl myristate reverse micelles as transdermal insulin carriers: Experimental evaluation and molecular dynamics simulation | |
| CN104083326B (en) | A kind of preparation method of the liposome wrapping load protein medicaments | |
| CN1242740C (en) | Mitoxantrone or mitoxantrone hydrochloride liposome injection and its preparing process | |
| CN100336507C (en) | Nimoldipine new nano liposome, its precursor freeze dryed matter and its preparing method | |
| CN1907266A (en) | Potassium dehydroandrographolide succinate/potassium dehydroandrographolide succinate liposome composition and production method thereof | |
| Rani et al. | Liposomal-Based Pharmaceutical Formulations–Current Landscape, Limitations and Technologies for Industrial Scale-Up | |
| CN1211073C (en) | Process for preparing ceramide liposome emulsion | |
| WO2016101854A1 (en) | Icg-encapsulated polymer-phospholipid nanoparticle and preparation method therefor | |
| CN103536533B (en) | A kind of water-soluble drug liposome preparation method | |
| WO2016105869A1 (en) | Method for making liposomes containing an active pharmaceutical ingredient | |
| CN105395485B (en) | A kind of bicyclic alcohols liposome and preparation method thereof | |
| CN1543931A (en) | Method for preparing brain targeting liposome pharmaceutical | |
| CN215350808U (en) | Injection type liposome medicine carrying equipment | |
| CN1257714C (en) | Injection of liposome of daunorubicin or hydrochloric daunorubicin and preparationmethod |
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 |
Granted publication date: 20050126 Termination date: 20150917 |
|
| EXPY | Termination of patent right or utility model |