CN102935068B - Preparation method of liposome entrapping water-soluble medicines - Google Patents
Preparation method of liposome entrapping water-soluble medicines Download PDFInfo
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Abstract
The invention relates to a preparation method of a liposome entrapping water-soluble medicines. According to the invention, a water-soluble medicine is dissolved in amino-acid-structure-containing high-molecular hydrophilic colloids such as gelatin, collagen or albumin; poloxamer is added and the mixture is well mixed, such that a W/W type colloidal solution is formed; the solution is lyophilized; the formed lyophilized powder is transferred into a tert-butanol solution containing a liposome material, and the mixture is well dispersed; and the mixture is subjected to lyophilization, such that high-entrapment-efficiency liposome entrapping the water-soluble medicine is formed. According to the liposome preparation method, W/W type colloid is combined with a two-step lyophilization preparation technology, such that problems such as low water-soluble medicine load, low entrapment efficiency and poor stability of existing preparation methods of liposome entrapping water-soluble medicines are solved. The method provided by the invention is used for preparing liposome used for entrapping water-soluble medicines, and is especially suitable for preparing liposome used for entrapping poor-thermal-stability or macromolecular medicines. The method can also be used for covering medicine bitterness or odor, and for separating medicines from other components.
Description
[technical field]
The invention belongs to pharmaceutics field, more particularly, the present invention relates to a kind of liposome and preparation method thereof.
[background technology]
Liposome (1iposomes) is a kind of single or multiple lift microcapsule being made up of the orderly lipid bilayer of arrangement.Liposome belongs to colloid system, has the cytoid structure of class, strong with cell membrane affinity, can increase the ability of encapsulated medicine permeate through cell membranes.Liposome good biocompatibility, can realize targeted delivery in medicine body, there is prolong drug action time, increase medicine inside and outside stability, reduce drug toxicity, strengthen the plurality of advantages such as pharmacological action.
Desirable liposome need to reach following requirement: (1) liposome form rounding and not assembling, can effectively control particle size range by preparation method, and realize the object of slow-releasing and controlled-releasing action and the targeted delivery of medicine; (2) liposome stability is high, can place for a long time; (3) liposome has higher envelop rate and drug loading, especially for poor heat stability medicine or water-soluble biological macromolecular drug; (4) liposome is easily realized sterilizing or sterile working.
The preparation method of liposome has multiple, as mechanical dispersion method, film dispersion method, reverse phase evaporation, multi-emulsion method, fusion method, injection method, freeze-drying, surfactant facture, calcium fusion method, carrier deposit method etc.Wherein, reverse phase evaporation, be the better method of water soluble medicament-entrapping in conjunction with the freeze-drying of thawing law technology repeatedly.But, still Shortcomings of the method for preparing lipidosome of existing water soluble medicament-entrapping, for example reverse phase evaporation needs long heat treatment process, is unsuitable for the protein and peptide drugs of poor heat stability, and in preparation process, organic solvent is easily residual, and potential hazard is large.Easily realize asepticize operation in conjunction with thawing law technology freeze-drying method repeatedly, the lipid freeze-dry powder good stability obtaining, but while meeting water formation liposome solutions in conjunction with the lipid freeze-dry powder that the freeze-drying method of thawing law technology prepares repeatedly, particle diameter increases several times conventionally, form is rounding and easily gathering not, cannot effectively control particle size range, have safety issue.
Patent of invention " a kind of new method of preparing liposome " (application number 03111470.9) discloses a kind of new method of preparing liposome; its claim 1 is: " a kind of new method of preparing liposome; it is characterized in that: a. prepares a single phase soln; its solute is: (1) is used to form the lipid of liposome, material to be encapsulated; or (2) be used to form the lipid of liposome, material and water-solubility carrier to be encapsulated, and described water-solubility carrier is sucrose, lactose or mannitol; Its solvent is made up of the tert-butyl alcohol and water, and the volume ratio of the tert-butyl alcohol and water is greater than 1:3.B. freezing single phase soln is removed solvent, obtains lyophilized products, and the lyophilized products aquation obtaining is obtained to liposome ".
Patent of invention " a kind of new method of preparing liposome " (application number 03111470.9), although utilized the freeze drying process that the tert-butyl alcohol is solution, is clearly emphasized: (1) water-solubility carrier is sucrose, lactose or mannitol; (2) before lyophilization, must obtain single phase soln system.Although the lipid freeze-dry powder that this patent of invention prepares is dissolved in the water, the liposome particle diameter forming is little, but structure is still traditional unilamelar liposome, because easily seeing through lipid film, the micromolecular water soluble substances such as sucrose, lactose or mannitol are dissolved in dissolution medium.In addition the liposome that prepared by this invention is the rear hyperosmosis that produces because inner micromolecular water solubleness carrier dissolves, therefore there is the problem of the quick seepage of water soluble drug, and this invention entrapment efficiency is lower, and (embodiment 5 is 21%, embodiment 6 is 40%), cannot meet Chinese Pharmacopoeia 2010 editions for the liposome encapsulation requirement of (being greater than 85%).
Patent of invention CN201210022488.9 discloses brood lac core lipid body lyophilized powder and a preparation method, this liposome has the similar structure of the present invention, but preparation method is that 1 part of high molecular weight hydrophilic colloidal materials is dissolved in 10-200 part water, add 0.05-50 part water soluble drug, mix formation water; 0.5-50 part matrix material, 2-100 part emulsifying agent and 10-1000 part frozen-dried supporting agent are dissolved in 50-4000 part tert-butyl alcohol, form oil phase; Water is distributed to and in oil phase, forms w/o type emulsion, and ultrasonic, high-speed stirred or high pressure homogenize are processed and are formed w/o type microemulsion solution, adopt super low temperature quick frozen technique to form solid dispersion, and the tert-butyl alcohol is removed in lyophilization, forms lipid freeze-dry powder.Patent of invention CN201010131339.7 discloses a kind of preparation method of wrapping medicine carrying composite lipidosome: by water soluble drug and amphipathy macromolecule material dissolves in aqueous phase system, carry out lyophilization processing for the first time, then the lyophilized powder that comprises water soluble drug and Liposome film-forming material are dispersed in organic facies system, carry out freeze-drying method for the second time, make the liposome of water soluble medicament-entrapping.Patent of invention CN201010131244.5 discloses a kind of preparation method of wrapping medicine carrying composite lipidosome: medicine is scattered in the amphipathy macromolecule material system of dissolving or molten condition, rapid cooling processing obtains medicine solid dispersion, then medicine solid dispersion and Liposome film-forming material are dispersed in organic facies system, according to method for preparing lipidosome, make the liposome of bag medicine carrying thing.Above three patents of invention, part has solved the problem such as poor stability or complicated operation existing in the preparation technology of liposome entrapment medicine, but operating time control is had relatively high expectations or needs heating and melting processing etc., increase the destruction to macromolecular drug, increased the operation control difficulty of preparation in batches simultaneously.
In a word, the method for preparing lipidosome of existing water soluble medicament-entrapping adopts Emulsion system or double emulsion system mostly, and need to pass through long heat treatment process, due to features such as the easy gatherings of macromole water soluble drug poor heat resistance, oil-water interfaces, the liposome of water soluble medicament-entrapping prepared by these class methods of application can not ensure the activity of the activity, particularly macromolecular drug of medicine.In addition, the liposome of water soluble medicament-entrapping prepared by these class methods exists that microgranule is easily assembled, drug delivery amount is low, envelop rate is low, the problems such as burst effect is obvious, adopt drug delivery technologies initiatively can appropriateness to improve the envelop rate of medicine, the steps such as desalination but active drug delivery technologies need to be dialysed, operating time length and poor reproducibility, be also unsuitable for the extensive preparation of the liposome of water soluble drug.
[summary of the invention]
The technical problem to be solved in the present invention is the weak point for existing method for preparing lipidosome, a kind of liposome technology of preparing that is suitable for water soluble medicament-entrapping is provided, water soluble drug is dissolved in to gelatin, collagen or albumin etc. containing in the high molecular weight hydrophilic colloid of amino acid structure, add poloxamer (Poloxamer) to mix rear formation W/W type colloid solution, lyophilization, the lyophilized powder forming proceeds in the t-butanol solution that contains matrix material, lyophilization again after being uniformly dispersed, forms the liposome of the water soluble medicament-entrapping of high envelop rate.This method for preparing lipidosome utilizes secondary freeze drying technology to solve that the method for preparing lipidosome of existing water soluble medicament-entrapping is little for water soluble drug drug loading, envelop rate is low and the problem of poor stability, the drug-loaded liposome particle diameter preparing in nanoscale, be uniformly dispersed, drug loading is large, entrapment efficiency is high, drug release is slow.The method is suitable for the liposome of preparing water soluble medicament-entrapping, be particularly useful for the liposome of the macromolecular drugs such as the medicine of preparation bag heat-carrying poor stability and albumen, polypeptide, polysaccharide, reach the object of slow controlled release drug administration or targeted delivery administration, also can for cover bitterness or stink, with other set isolations from, reduce medicine irritation effect, liquid medicine is converted to the preparation objects such as solid form, can be applicable to injection, oral, mucosa, skin, wound surface, the part of tract or the multiple dosage form of whole body therapeutic.
Inventor finds in long term test: 1. gelatin, collagen or albumin etc. have good ability to arrange jobs containing the high molecular weight hydrophilic colloid solution of amino acid structure for water soluble drug, but be difficult to long-term preservation, after mixing containing the high molecular weight hydrophilic colloid of amino acid structure and poloxamer (Poloxamer), can form W/W type colloid solution, macromolecular drug is easily scattered in the interior water forming containing the high molecular weight hydrophilic colloid solution of amino acid structure; 2. the lyophilized powder that utilizes lyophilization that W/W type colloid solution is made is conducive to the stable of medicine; 3. poloxamer (Poloxamer) is good lyophilization proppant, and the liposome stability that freeze-dry process obtains is high; 4. the lyophilized powder that W/W type colloid solution is made is scattered in that the liposome particle diameter forming in phospholipid solution is little and stability is high again.Inventor is through great many of experiments, innovatively " W/W type colloid solution technology " and " secondary freeze drying technique " organically blended integral, the features such as the high stability of the integrated high drug load of colloid, freeze drying process, explore a kind of liposome technology of preparing that is suitable for water soluble medicament-entrapping, the problem that overcomes that the drug loading that traditional liposomal water soluble medicament-entrapping exists is low, entrapment efficiency is low, burst effect is obvious etc.
The key technology of method for preparing lipidosome of the present invention is combined as: 1. preparation mixes rear formation W/W type colloid solution containing high molecular weight hydrophilic colloid and the poloxamer (Poloxamer) of amino acid structure, and macromolecular drug is easily scattered in the interior water containing the high molecular weight hydrophilic colloid solution of amino acid structure; 2. adopt freeze drying process that the interior W/W type macromolecule glue liquid solution that holds mutually macromolecular drug is made to lyophilized powder, be conducive to keep the high degree of dispersion state of medicine and maintain stability; 3. the hydrophilic colloid lyophilized powder that holds water soluble drug proceeds in the t-butanol solution that contains matrix material, utilize ultrasonic technique to make phospholipid material fully wrap up the microgranule that hydrophilic colloid is core, form suspension solution system, and by regulating ultrasound intensity or tert-butyl alcohol consumption control particle size; 5. secondary freeze drying, forms the lipid freeze-dry powder of the water soluble medicament-entrapping of high envelop rate, injects before use solvent for injection, forms the liposome solutions of water soluble medicament-entrapping, solves the problem of macromolecular drug poor stability.
Taking bovine serum albumin as example, experimental group: according to Mass Calculation, 1 part of gelatin is dissolved in 1000 parts of water, adds 5 parts of bovine serum albumin and 100 parts of Poloxamer 188, mix and form hydrophilic colloid solution I, lyophilization, forms lyophilized powder I.5 parts of hydrogenation egg yolk lecithin, 10 parts of polyoxyethylene hydrogenated Oleum Ricini are dissolved in 200 parts of tert-butyl alcohols, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in liquid nitrogen, and lyophilization forms the lipid freeze-dry powder of bag year bovine serum albumin.
Inventor is provided with controlled trial and observes, and controlled trial group is as follows:
Controlled trial group 1(does not add gelatin): according to Mass Calculation, 5 parts of bovine serum albumin and 200 parts of Poloxamer188 are dissolved in to 1000 parts of water, mix, form hydrophilic colloid solution I, lyophilization, forms lyophilized powder I.5 parts of hydrogenation egg yolk lecithin, 10 parts of polyoxyethylene hydrogenated Oleum Ricini are dissolved in 200 parts of tert-butyl alcohols, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in liquid nitrogen, and lyophilization forms the lipid freeze-dry powder of bag year bovine serum albumin.
In controlled trial group 2(hydrophilic colloid, do not add poloxamer): according to Mass Calculation, 1 part of gelatin is dissolved in 1000 parts of water, adds 5 parts of bovine serum albumin, mix and form hydrophilic colloid solution I, lyophilization, forms lyophilized powder I.5 parts of hydrogenation egg yolk lecithin, 10 parts of polyoxyethylene hydrogenated Oleum Ricini are dissolved in 200 parts of tert-butyl alcohols, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in liquid nitrogen, and lyophilization forms the lipid freeze-dry powder of bag year bovine serum albumin.
Only primary freeze drying processing of controlled trial group 3(): according to Mass Calculation, 1 part of gelatin is dissolved in 1000 parts of water, adds 5 parts of bovine serum albumin and 200 parts of Poloxamer 188, mix and form hydrophilic colloid solution I.5 parts of hydrogenation egg yolk lecithin, 10 parts of polyoxyethylene hydrogenated Oleum Ricini are dissolved in 200 parts of tert-butyl alcohols, form t-butanol solution I.Hydrophilic colloid solution I is joined in t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in liquid nitrogen, and lyophilization forms the lipid freeze-dry powder of bag year bovine serum albumin.
Controlled trial group 4, prepares liposome, concrete grammar according to patent of invention CN201210022488.9 method: according to Mass Calculation, 1 part of gelatin is dissolved in 1000 parts of water, adds 5 parts of bovine serum albumin to mix formation water; 5 parts of hydrogenation egg yolk lecithin, 10 parts of polyoxyethylene hydrogenated Oleum Ricini and 200 parts of Poloxamer 188 are dissolved in 500 parts of tert-butyl alcohols, form oil phase; Water is distributed in oil phase and forms w/o type emulsion, and 80KHz supersound process 2min forms w/o type microemulsion solution, adopts liquid nitrogen flash freezer technique to form solid dispersion, and the tert-butyl alcohol is removed in lyophilization, forms lipid freeze-dry powder.
Controlled trial group 5, prepares liposome, concrete grammar according to patent of invention CN201010131339.7 method: according to Mass Calculation, 200 parts of polyvidones are dissolved in 1000 parts of water, add 5 parts of bovine serum albumin to dissolve ,-30 DEG C freezing 5 hours, lyophilization obtains solid-state dried frozen aquatic products; 5 parts of hydrogenation egg yolk lecithin, 2 parts of cholesterol, 0.5 part of Tween 80 are dissolved in 500 parts of tert-butyl alcohols, form oil phase; Above-mentioned solid-state dried frozen aquatic products is distributed in oil phase, adds 200 portions of mannitol, ultrasonic (80KHz) processes 2min and makes to be uniformly dispersed, and lyophilization obtains lipid freeze-dry powder.
Controlled trial group 6, prepares liposome, concrete grammar according to patent of invention CN201010131244.5 method: according to Mass Calculation, 200 parts of polyvidone meltings in 65 DEG C of water-baths, add 5 parts of bovine serum albumin to be uniformly dispersed, go to quenching processing in 0 DEG C of ice bath, form solid dispersion; 5 parts of hydrogenation egg yolk lecithin, 2 parts of cholesterol, 0.5 part of Tween 80 are dissolved in 500 parts of tert-butyl alcohols, form oil phase; Above-mentioned solid dispersion is distributed in oil phase, adds 200 portions of mannitol, ultrasonic (80KHz) processes 2min and makes to be uniformly dispersed, and lyophilization obtains lipid freeze-dry powder.
Found that, the bovine serum albumin lipid freeze-dry powder of experimental group is dissolved in 2000 parts of waters for injection, liposome particle diameter less (mean diameter=810nm), and microscopic pattern rounding, is uniformly dispersed, and places and within 24 hours, there is no significant change (Fig. 1).Adopt polydextran gel partition method in conjunction with bovine serum albumin specific detection kit measurement envelop rate, the bovine serum albumin liposome encapsulation of experimental group reaches 95.4%, and measures after placing 30min, and bovine serum albumin liposome does not have burst effect.
Controlled trial group 1(does not add macromolecule hydrophilic colloid) bovine serum albumin lipid freeze-dry powder be dissolved in 2000 parts of waters for injection, liposome initial particle is less, but aggregation velocity is fast, (>2 μ m) easily to form macroparticle, initial envelop rate reaches 86.2%, but drug leakage is serious, there is obvious burst effect.
In controlled trial group 2(hydrophilic colloid, do not add poloxamer) bovine serum albumin lipid freeze-dry powder be dissolved in 2000 parts of waters for injection, particle diameter is large, and (>2 μ m), shape is rounding not, and skewness, and liposome encapsulation is 89.6%.
Only primary freeze drying processing of controlled trial group 3() bovine serum albumin lipid freeze-dry powder be dissolved in 2000 parts of waters for injection, liposome particle diameter is large, and (m), shape is rounding not for >2 μ, and liposome encapsulation is 70.2%.
Controlled trial group 4(patent of invention CN201210022488.9 method) in being distributed to oil phase, water forms in the preparation manipulation of w/o type emulsion, and must adopt the supersound process of higher-wattage could form w/o type microemulsion solution.The bovine serum albumin lipid freeze-dry powder preparing is dissolved in 2000 parts of waters for injection, liposome particle diameter large (mean diameter=980nm), and not rounding very of micro-shape, liposome encapsulation is 88.2%.
Controlled trial group 5(patent of invention CN201010131339.7 method) solid-state dried frozen aquatic products is distributed in the preparation manipulation in oil phase, must adopt the supersound process of higher-wattage just can make Solution Dispersion even.The bovine serum albumin lipid freeze-dry powder preparing is dissolved in 2000 parts of waters for injection, liposome particle diameter large (mean diameter=1.1 μ m), not rounding very of micro-shape, liposome encapsulation is 86.1%.
Controlled trial group 6(patent of invention CN201010131244.5 method) solid dispersion being distributed in the preparation manipulation in oil phase, must adopt the supersound process of higher-wattage just can make Solution Dispersion even, the bovine serum albumin lipid freeze-dry powder preparing is dissolved in 2000 parts of waters for injection, liposome particle size distribution range is wider, mean diameter is large, and (mean diameter=1.5 μ m), it is irregularly shaped that micro-shape is, and liposome encapsulation is 85.7%.
Experimental result shows, the key technology combination of method for preparing lipidosome of the present invention is indispensable.Contrast prior art, " W/W type colloid solution technology " and " secondary freeze drying technique " combination technology of adoption of innovation of the present invention, ensure better the activity of high molecular weight protein class medicine, improve the controllability of operation, reduce the difficulty of processing procedure, especially reduced and formed the required mechanical treatment intensity (as ultrasonic time and power) of w/o type microemulsion solution, also improved drug-loaded liposome rounding property and envelop rate.
In addition, the tert-butyl alcohol amount in the poloxamer consumption in the hydrophilic colloid solution I of experimental group and the water yield, t-butanol solution I is increased by 1 times by inventor, all the other steps are constant, the liposome mean diameter that the bag obtaining carries bovine serum albumin is 550nm, show, by the adjusting of poloxamer consumption or liquor capacity in preparation process, can control the particle diameter of liposome.
In addition, different model or the consumption of some components of experimental group do not exert an influence to experimental result, and for example phospholipid material adopts hydrogenated phospholipid or synthetic phospholipid, and particle diameter and the envelop rate of the drug-loaded liposome of formation are consistent; Poloxamer raw material adopts the different models such as Poloxamer 188 or Poloxamer 407, and particle diameter and the envelop rate of the drug-loaded liposome of formation are consistent; Bovine serum albumin lipid freeze-dry powder prepared by experimental group is dissolved in 1000 parts of-50000 parts of waters for injection, and particle diameter and the envelop rate of the drug-loaded liposome of formation are consistent.Therefore, can be according to clinical practice needs, select suitable phospholipid material, poloxamer model or the injection water yield.
In experiment, also find, traditional liposomal needs a large amount of phospholipid materials could wrap year certain medicine, medicine: the minimum 1:5 that is no less than of mass ratio of phospholipid, and medicine of the present invention: it is even lower that the mass ratio of phospholipid can reach 1:1, save the consumption of phospholipid, significantly improved the Drug loading capacity of liposome.In addition, preparation technology of the present invention is easy, does not need to adopt pH gradient or ammonium sulphate gradient can prepare high envelop rate drug-loaded liposome, saves the complicated operating process such as desalination and high pressure homogenize, easily realizes asepticize operation.
Thus, a kind of method for preparing lipidosome of water soluble medicament-entrapping, its preparation process comprises following process:
A. calculate with quality proportioning, add 500-5000 part water to form solution 1 part of high molecular weight hydrophilic glue material containing amino acid structure, add 0.5-50 part water soluble drug and 20-500 part poloxamer to dissolve, form the W/W type hydrophilic colloid solution that contains medicine, lyophilization, forms lyophilized powder I;
B. calculate with quality proportioning, by 1-50 part matrix material, 5-500 part polyoxyethylene hydrogenated Oleum Ricini, is dissolved in 500-10000 part tert-butyl alcohol, adopts the processing method accelerate dissolution of ultrasonic or heating in water bath, forms t-butanol solution I;
C. lyophilized powder I is joined in t-butanol solution I, be uniformly dispersed, form t-butanol solution II, in liquid nitrogen or refrigerator, freeze to form solid, lyophilization forms the lipid freeze-dry powder II of water soluble medicament-entrapping, fill seals preservation in cillin bottle, injects before use the known solvent for injection of 1000-50000 part pharmacy, forms the liposome solutions of water soluble medicament-entrapping.
The above-mentioned high molecular weight hydrophilic glue material containing amino acid structure comprises gelatin, collagen and human albumin.
Above-mentioned water soluble drug refers to independent application, coordinates solubilizing agent application or the diagnostic reagent of drug solubility is greater than 0.01mg/ml within the scope of specific pH value biopharmaceutical macromolecular drug, chemicals, Chinese medicine effective extract, medical science or field of biology application.
Above-mentioned biopharmaceutical macromolecular drug refers to medicine and their biodegradation or the derivative products of protein, polypeptide, polysaccharide, enzyme, coenzyme, nucleic acid structure, comprises the polysaccharide of somatomedin, hormone, stimulating factor, antibody, vaccine, interferon, interleukin, plant and animal material.
Above-mentioned water soluble drug comprises: insulin, hirudin, vascular endothelial growth inhibitive factor, neurotrophic factor, cell growth factor, osteogenic growth factor, breast iron transfer albumen, interferon, fluorescin, oxaliplatin, carboplatin, nedaplatin, aclarubicin, doxorubicin, epirubicin, mitoxantrone, bleomycin, Bleomycin A5, mitomycin, Erlotinib, gefitinib, imatinib, Dasatinib, alizapride, azasetron, ondansetron, chlorine phosphoric acid, mesna, Rituximab, ibritumomab tiuxetan, Cetuximab, bevacizumab, Anastrozole, aminoglutethimide, formestane, exemestane, teniposide, etoposide, pentostatin, irinotecan, busulfan, chlormethine, Ka Mosiding, Lomustine, homoharringtonine, asparaginase, pegaspargase, cytosine arabinoside, floxuridine, gemcitabine, AZD2171, alendronate, Rosuvastatin, antithrombase, carmofur, docetaxel, vindesine, vincristine, paclitaxel, BAY 43-9006, decitabine, procarbazine, nimodipine, nifedipine, nitrendipine, felodipine, diclofenac, naproxen, tramadol hydrochloride, morphine, nitroglycerin, clonidine, Ismo 20, ticlopidine, acetazolamide, acetaminophen, aminophylline, amitriptyline, ampicillin, amoxicillin, aspirin, beclamide, caffeine, cimetidine, phenobarbital, Camphora, chloromycetin, Chlophenamin, chlorpromazine hydrochloride, CLOF, cloxacillin, codeine phosphate, diazepam, dextromethorphan, ibuprofen, diphhydramine hydrochloride, doxycycline hydrochloride, eprazinone, fenfluramine, ferrous citrate, ferrous fumarate, ferrous sulfate, scopolamine, pseudoephedrine, berberine, indomethacin, levodopa, lithium carbonate, meclofenoxate hydrochloride, methaqualone, methyl An Feitaiming, acetylspiramycin, nitrofurantoin, nortriptyline, narcotine, hydrochloric acid handkerchief is exerted Lamine, phenacetin, Phenylbutazone, hydrochloric acid benzene good fortune is bright, hydrochloric acid amfetamine alcohol, prednisolone, procainamide, propantheline bromide, Doxaphene, Propranolol, sulphuric acid Kui Nier, thioridazine, zinc gluconate, sulfametomidine, tetracycline, streptomycin, gentamycin, trifluomeprazine, alimemazine, vitamin, nuclear-magnetism contrast agent, isotope.
Above-mentioned matrix material refers to the known phospholipid of pharmacy and lipid materials, comprises natural phosphatidyl choline, hydrogenated phospholipid, synthetic phospholipid and polyethyleneglycol modified derivant thereof, saturated fatty acid glyceride, steroidal.
In the hydrophilic colloid solution that contains medicine of above-mentioned preparation process a, can also add 0.1-50 part pharmacy generally acknowledged water solublity antioxidant, stabilizing agent, pH adjusting agent, wherein stabilizing agent comprises: human serum albumin, tartaric acid, succinic acid, cholic acid, deoxycholic acid, fumaric acid, citric acid, Palmic acid, aminoacid.
In the t-butanol solution I of above-mentioned preparation process b, can also add 0.1-50 part pharmacy generally acknowledged oil-soluble antioxidant, organic acid and ester thereof, wherein organic acid comprises: formic acid, acetic acid, tartaric acid, Palmic acid, stearic acid, oleic acid.
The method for preparing lipidosome of above-mentioned a kind of water soluble medicament-entrapping, it is characterized in that: the drug-loaded liposome lyophilized powder of preparation injects the known solvent for injection of 1000-50000 part pharmacy before use as required, form the drug-loaded liposome of particle diameter within the scope of 100nm-1000nm.
Above-mentioned liposome is by selecting dissimilar phospholipid or lipid materials combination, make the liposome of long circulating liposomes, pH sensitive liposome body that PEG modifies, responsive to temperature liposome, surface band positive charge, bring into play better in gastrointestinal tract blood circulation targeting drug release effect in positioning release medicine or body.
Above-mentioned liposome is to use separately or be combined in preparation prescription to apply, by injection, oral, mucosa, skin, wound surface, tract administration, performance treatment, diagnosis, prevention, immunity, clean, the application of sterilization, beauty treatment, health care.
The method for preparing lipidosome of a kind of water soluble medicament-entrapping of the present invention, there is following advantage: (1) this liposome preparation technology organically blends integral by " W/W colloid technology " and " secondary freeze drying technique " innovatively, the features such as the high stability of the high drug load of integrated colloid and polymolecularity, freeze drying process, avoid medicine to be heated, realized high degree of dispersion and the particle size uniformity of the liposome of bag medicine carrying thing; (2) this liposome preparation technology has effectively improved the efficiency of traditional liposomal freeze-drying preparation technology, reduces lyophilizing Operating Complexity.(3) medicine of liposome of the present invention: it is even lower that the mass ratio of phospholipid can reach 1:1, has saved the consumption of phospholipid, significantly improves the Drug loading capacity of liposome; (4) preparation technology of the present invention is easy, does not need to adopt pH gradient or ammonium sulphate gradient can prepare high envelop rate liposome, saves the complex operations processes such as dialysis desalination, easily realizes asepticize operation; (5) can be by regulating hydrophilic colloid proportioning, tert-butyl alcohol consumption etc. to control liposome particle diameter; (6) the liposome particle diameter that prepared by the present invention is in nanometer range, and can effectively control particle diameter, be conducive to biomembrane barrier in medicine transdermal, mucosa or body, applied widely, can be applicable to multiple dosage form, meet injection, oral, mucosa, skin, wound surface, the part of tract or the needs of whole body therapeutic.(7) liposome of the present invention can be used as slow controlled release drug delivery system or targeted delivery drug-supplying system, also can for cover bitterness or stink, with other set isolations from, reduce medicine irritation effect, liquid medicine is converted to the preparation objects such as solid form.
[brief description of the drawings]
Fig. 1: bag carries the microscopic pattern (mean diameter=810nm) of bovine serum albumin liposome
Fig. 2: water soluble medicament-entrapping liposome preparation flow schematic diagram
[detailed description of the invention]
Now further describe the present invention in conjunction with following example.
Embodiment 1: insulin liposome
The present embodiment is prepared insulin liposome, and insulin liposome forms as table 1.
Table 1 insulin liposome component proportion
Insulin liposome method for preparing freeze-dried powder: shown in as shown in Fig. 2 flow process, according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add insulin, poloxamer (Poloxamer), additives I to dissolve, the hydrophilic colloid solution that formation contains medicine, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in 60 DEG C of tert-butyl alcohols, mix and form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and the ultrasonic 2min of 20KHz, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of water soluble medicament-entrapping, and fill seals preservation in cillin bottle.
Granularmetric analysis: get the lipid freeze-dry powder 100mg that each prescription obtains, add 2ml water for injection to form liposome solutions, you measure mean diameter by special particle size analyzer application library.
Result: prepared liposome, the insulin liposome mean diameter of prescription 1 is 915nm, the insulin liposome mean diameter of prescription 2 is 742nm, and the insulin liposome mean diameter of prescription 3 is 286nm, and the insulin liposome mean diameter of prescription 4 is 220nm, the insulin liposome mean diameter of prescription 5 is 120nm, the insulin liposome particle diameter of prescription 6-11 is respectively: 970nm, 290nm, 950nm, 265nm, 890nm and 280nm.Result shows to regulate the consumption of the tert-butyl alcohol and frozen-dried supporting agent can control liposome particle diameter.The consumption of the tert-butyl alcohol and frozen-dried supporting agent is larger, and the particle diameter of liposome is less.
Embodiment 2: growth hormone liposome
The present embodiment is prepared growth hormone liposome, and growth hormone liposome forms as table 2.
Table 2 growth hormone liposome component proportion
Growth hormone liposome method for preparing freeze-dried powder: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add growth hormone, poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains growth hormone, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and 55 DEG C of heating in water bath, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (60KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag carrying growth hormone, and fill seals preservation in cillin bottle.
Embodiment 3: the quality evaluation of growth hormone liposome
The present embodiment utilizes growth hormone liposome prepared by embodiment 2 to carry out quality evaluation.
Microscopic pattern and granularmetric analysis: get 100mg lipid freeze-dry powder and add 2ml water to form liposome solutions, draw respectively a certain amount of liposome turbid liquor also with 1% phosphotungstic acid dyeing, be placed in the morphological characteristic of observing liposome under scanning electron microscope, you measure liposome particle diameter by special particle size analyzer application library.
Entrapment efficiency determination: get 1ml liposome solutions, be splined on Sephadex G-50 gel column, taking distilled water as eluent, access the eluting part of different volumes, separate and receive free growth hormone eluting part, utilize HPLC method to analyze content, adopt " envelop rate (%)=[(growth hormone total amount-free growth hormone amount)/growth hormone total amount] × 100 " formula to calculate the envelop rate of growth hormone liposome.
Experimental result is in table 3, and growth hormone liposome prepared by visible the present invention has good particle size distribution and microscopic pattern, envelop rate high (all exceeding 90%), and without burst effect.In addition the obvious positive charges of growth hormone liposome surface band (surface potential approximately+42mv) of prescription 3 preparations, the faint negative charge of the surface of liposome band of other prescription (surface potential approximately-16mv).
The quality evaluation of table 3 growth hormone liposome
Note: according to Pharmacopoeia of People's Republic of China (2010 editions) regulation, the microgranules such as liposome, microsphere are greater than 40% in the burst size starting in 0.5h, have burst effect.
Embodiment 4: the preparation of hydrochloric doxorubicin liposome
The present embodiment is prepared hydrochloric doxorubicin liposome, and hydrochloric doxorubicin liposome forms as table 4.
Table 4 hydrochloric doxorubicin liposome component proportion
Note: DPPC is dipalmitoyl phosphatidyl choline; DSPC is phosphide distearoyl phosphatidylcholine; MSPC is myristoyl stearoyl lecithin; DSPE-PEG2000 is the phosphide distearoyl phosphatidylcholine that PEG2000 modifies.
Hydrochloric doxorubicin liposome method for preparing freeze-dried powder: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add doxorubicin hydrochloride, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains doxorubicin hydrochloride, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and the ultrasonic 30s of 90KHz, forms t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and the ultrasonic 1min of 50KHz, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag year doxorubicin hydrochloride, and fill seals preservation in cillin bottle.
Prepared by prescription 2 and 3 is long circulating liposomes, and prepared by prescription 4 and 5 is temperature sensitive liposome.
Matched group 1--film dispersion method is prepared hydrochloric doxorubicin liposome: take hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in pear shape bottle with 20ml ether dissolution, rotary evaporation film forming, and continue decompression rotary evaporation 15min and eliminate to ether, add 1mg/ml doxorubicin hydrochloride solution 10ml, after aquation 1h, ultrasonic 15min under Probe Ultrasonic Searching, crosses 0.22 μ m filter membrane 4 times, granulate, obtain hydrochloric doxorubicin liposome solution, add mannitol 1g, lyophilization, obtains hydrochloric doxorubicin liposome lyophilized powder.
Matched group 2--ammonium sulphate gradient is prepared hydrochloric doxorubicin liposome lyophilized powder: take hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in pear shape bottle with 20ml ether dissolution, add the ammonium sulfate 10ml of 250mmol/L, ultrasonic 3min becomes colostrum, decompression rotary evaporation 20min eliminates to ether, ultrasonic 15min under Probe Ultrasonic Searching, cross 0.22 μ m filter membrane 4 times, granulate, obtain blank liposome liquid 10ml, remove the salt in the outer water of this blank liposome, add wherein again doxorubicin hydrochloride drug powder 10mg, stir after 2min, leave standstill 20min, obtain hydrochloric doxorubicin liposome solution, add mannitol 1g, lyophilization, obtain hydrochloric doxorubicin liposome lyophilized powder.
Matched group 3--reverse phase evaporation is prepared hydrochloric doxorubicin liposome lyophilized powder: take hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in pear shape bottle with 20ml ether dissolution, add 1mg/ml doxorubicin hydrochloride aqueous solution 10ml, ultrasonic 3min becomes colostrum, decompression rotary evaporation 20min eliminates to ether, ultrasonic 15min under Probe Ultrasonic Searching, be pressed through 0.22 μ m filter membrane 4 times, obtain hydrochloric doxorubicin liposome solution, add mannitol 1g, lyophilization, obtains Evacet lyophilized powder.
Embodiment 5: the quality evaluation of hydrochloric doxorubicin liposome
The present embodiment utilizes the prescription of embodiment 4 and hydrochloric doxorubicin liposome prepared by matched group to carry out quality contrast.
Microscopic pattern and granularmetric analysis: with embodiment 3.
Entrapment efficiency determination: get 100mg lipid freeze-dry powder and add 2ml water to form liposome solutions, be splined on SephadexG-50 gel column, taking distilled water as eluent, access the eluting part of different volumes, separate the liposome eluting part that receives bag year doxorubicin hydrochloride, add Triton-100 to destroy hydrochloric doxorubicin liposome, extract amycin and make amycin solution, utilize HPLC method to detect amycin content (chromatographic condition: Venusil MP C18 post (416mm × 250mm, m), mobile phase is acetonitrile-methanol-10mmolL to 5 μ
-1phosphate buffer (36: 32: 32), it is 230nm that UV detects wavelength, flow velocity is 1.0mL/min).Adopt " envelop rate (%)=[(the doxorubicin hydrochloride amount that bag carries)/doxorubicin hydrochloride total amount] × 100 " formula to calculate the envelop rate of hydrochloric doxorubicin liposome.
Drug release behavior is measured: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, pack in the bag filter of molecular cut off 3500, two ends fasten, and the method that application digestion instrument specifies according to 2010 editions pharmacopeia is measured release.Dissolution medium is the alcoholic solution 800ml of 0.25% sodium lauryl sulphate and 10%, temperature is 37 ± 0.5 DEG C, and mixing speed is 100r/min, timing sampling 5ml, and supplementing in time isothermal equal-volume blank medium, after sample 0.22 μ m filtering with microporous membrane, HPLC measures peak area.The each time point hydrochloric doxorubicin liposome of calculation sample Cumulative release amount.Prescription 4 and 5 is responsive to temperature liposome, and drug release behavior assay method is the same, but dissolution medium Temperature Setting is 37 DEG C, 40 DEG C and 43 DEG C of three levels, and to write out a prescription 1 as contrast.
Experimental result is in table 5: the hydrochloric doxorubicin liposome form rounding of 7 formula preparations of the present invention, without clustering phenomena, mean diameter is little and particle size distribution is narrower, experimental group envelop rate reaches more than 90%, far above matched group, the prepared long circulating liposomes of prescription 1-3, prescription 4 and 5 is responsive to temperature liposome.Drug release experiment shows, prescription 1,2,3,6 and 7 has good slow releasing function than matched group, be greater than 8h release time, prescription 4 and 5 responsive to temperature liposome in the time of 37 DEG C 24h cumulative release amount between 80-100%, in the time that temperature is brought up to 40 DEG C or 43 DEG C, rate of release is obviously accelerated, when 43 DEG C of 4h, burst size approaches 100%, therefore write out a prescription 4 and prescription the 5 responsive to temperature liposomees that prepare there is good thermo-sensitive property, be applicable to the cancer target administration in conjunction with thermotherapy.
The quality evaluation of table 5 hydrochloric doxorubicin liposome
Embodiment 6: the preparation of topotecan hydrochloride liposome and drug release behavior are measured
The lipidosome freeze-dried powder, preparation method thereof of topotecan hydrochloride: the prescription composition of topotecan hydrochloride liposome and preparation method are with the prescription 2 of embodiment 4, and the medicine amycin just bag being carried is changed to topotecan hydrochloride.
Drug release behavior is measured: with embodiment 5.
Experimental result: topotecan hydrochloride liposome form rounding, without clustering phenomena, mean diameter is 905nm, narrow diameter distribution, envelop rate reaches 93.2%.Drug release experiment shows, topotecan hydrochloride liposome has write out a prescription 2 the similar drug release behavior of Evacet with embodiment 5, has good slow releasing function.
Embodiment 7: ganoderan liposome
The present embodiment is prepared ganoderan liposome, and ganoderan liposome forms as table 6.
The lipidosome freeze-dried powder, preparation method thereof of ganoderan: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add ganoderan, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains ganoderan, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and 50 DEG C of heating in water bath, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag year ganoderan, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: method is with embodiment 3.
Entrapment efficiency determination: get 100mg lipid freeze-dry powder and add 1ml water to form liposome solutions, be splined on SephadexG-50 gel column, taking distilled water as eluent, access the eluting part of different volumes, separate and receive free ganoderan eluting part, utilize sulfuric acid anthrone colorimetric method to measure free ganoderma polyoses content, adopt " envelop rate (%)=[(ganoderan total amount-free ganoderan amount)/ganoderan total amount] × 100 " formula to calculate the envelop rate of ganoderan liposome.
Result: the ganoderan liposome of three formula preparations has good particle diameter, and mean diameter is less than 1000nm, without clustering phenomena, the envelop rate of the ganoderan liposome of three formula preparations all exceedes 90%.
Table 6 ganoderan liposome component proportion
Embodiment 8:bFGF liposome
The present embodiment is prepared basic fibroblast growth factor (bFGF) liposome, and bFGF liposome forms as table 7.
Table 7bFGF liposome component proportion
The lipidosome freeze-dried powder, preparation method thereof of bFGF: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add bFGF, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains bFGF, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and ultrasonic (70KHz) 1min, forms t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag year bFGF, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the bFGF liposome of each formula preparation has good particle diameter, and mean diameter is less than 950nm, without clustering phenomena.In addition the obvious positive charges of bFGF surface of liposome band (surface potential approximately+32mv) of prescription 8 preparations, the faint negative charge of the surface of liposome band of other prescription (surface potential approximately-15mv).
Embodiment 9: recombinant human vascular endothelial inhibin liposome
The present embodiment is prepared recombinant human vascular endothelial inhibin liposome, and recombinant human vascular endothelial inhibin liposome composition is same as table 8.
Table 8 recombinant human vascular endothelial inhibin liposome component proportion
The lipidosome freeze-dried powder, preparation method thereof of recombinant human vascular endothelial inhibin: the lipidosome freeze-dried powder, preparation method thereof of recombinant human vascular endothelial inhibin: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add recombinant human vascular endothelial inhibin, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains recombinant human vascular endothelial inhibin, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, lyophilization 25h forms the lipid freeze-dry powder II of bag year recombinant human vascular endothelial inhibin, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the recombinant human vascular endothelial inhibin liposome of each formula preparation has good particle diameter, and mean diameter is less than 900nm, without clustering phenomena.
Embodiment 10: transforming growth factor β (TGF-β) liposome
The present embodiment is prepared transforming growth factor β (TGF-β) liposome, and TGF-β liposome forms as table 9.
Table 9TGF-β liposome component proportion
The lipidosome freeze-dried powder, preparation method thereof of TGF-β: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add TGF-β, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains TGF-β, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag year TGF-β, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the TGF-β liposome of each formula preparation has good particle diameter, without clustering phenomena, the TGF-β liposome mean diameter of prescription 1 and prescription 2 preparations is 768nm, the TGF-β liposome mean diameter 418nm of prescription 3 preparations.。
Embodiment 11: human interferon-alpha-2 b liposome
The present embodiment is prepared human interferon-alpha-2 b liposome, and human interferon-alpha-2 b liposome forms as table 10.
Table 10 human interferon-alpha-2 b liposome component proportion
The lipidosome freeze-dried powder, preparation method thereof of human interferon-alpha-2 b: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add human interferon-alpha-2 b, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains human interferon-alpha-2 b, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of the manned interferon alpha 2 b of bag, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the human interferon-alpha-2 b liposome of each formula preparation has good particle diameter, without clustering phenomena, the human interferon-alpha-2 b liposome mean diameter of prescription 1 and prescription 2 preparations is 863nm and 802nm, the human interferon-alpha-2 b liposome mean diameter 874nm of prescription 3 preparations.
Embodiment 12: the preparation of pseudoephedrine liposome
The present embodiment, taking Effective Component of Chinese Medicine pseudoephedrine as object, is prepared pseudoephedrine liposome, and preparation forms as table 11.
Table 11 pseudoephedrine liposome component proportion
The lipidosome freeze-dried powder, preparation method thereof of pseudoephedrine: according to the quality proportioning of above each component, take respectively component, high molecular weight hydrophilic colloidal materials is added to the water to formation solution, add pseudoephedrine, Poloxamer, additives I to dissolve, the hydrophilic colloid solution that formation contains pseudoephedrine, lyophilization, forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, form t-butanol solution I.Lyophilized powder I is joined in t-butanol solution I, and ultrasonic (50KHz) 1min, forms t-butanol solution II, and in liquid nitrogen, quick-freezing forms solid, and lyophilization 25h forms the lipid freeze-dry powder II of bag year pseudoephedrine, and fill seals preservation in cillin bottle.
Microscopic pattern and granularmetric analysis: get 200mg lipid freeze-dry powder and add 2ml water to form liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the pseudoephedrine liposome of each formula preparation has good particle diameter, without clustering phenomena, the pseudoephedrine liposome mean diameter of prescription 1 preparation is 887nm, the pseudoephedrine liposome mean diameter 386nm of prescription 2 and prescription 3 preparations.
In the above-described embodiments, only the present invention has been carried out to exemplary description, but those skilled in the art are reading after present patent application and can carry out various amendments to the present invention without departing from the spirit and scope of the present invention.
Claims (2)
1. a method for preparing lipidosome for water soluble medicament-entrapping, is characterized in that: its preparation process comprises following process:
A. calculate with quality proportioning, add 500-5000 part water to form solution 1 part of high molecular weight hydrophilic glue material containing amino acid structure, add 0.5-50 part water soluble drug and 20-500 part poloxamer to dissolve, form the W/W type hydrophilic colloid solution that contains medicine, lyophilization, forms lyophilized powder I;
B. calculate with quality proportioning, by 1-50 part matrix material, 5-500 part polyoxyethylene hydrogenated Oleum Ricini, is dissolved in 500-10000 part tert-butyl alcohol, adopts the processing method accelerate dissolution of ultrasonic or heating in water bath, forms t-butanol solution I;
C. lyophilized powder I is joined in t-butanol solution I, be uniformly dispersed, form t-butanol solution II, in liquid nitrogen or refrigerator, freeze to form solid, lyophilization forms the lipid freeze-dry powder II of water soluble medicament-entrapping, fill seals preservation in cillin bottle, injects before use the known solvent for injection of 1000-50000 part pharmacy, forms the liposome solutions of water soluble medicament-entrapping.
2. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, is characterized in that: the described high molecular weight hydrophilic glue material containing amino acid structure is selected from gelatin, collagen and human albumin.
3. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, is characterized in that: described water soluble drug refers to independent application, coordinates solubilizing agent application or the diagnostic reagent of drug solubility is greater than 0.01mg/ml within the scope of specific pH value biopharmaceutical macromolecular drug, chemicals, Chinese medicine effective extract, medical science or field of biology application.
4. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 3, it is characterized in that: described biopharmaceutical macromolecular drug refers to medicine and their biodegradation or the derivative products of protein, polypeptide, polysaccharide, enzyme, coenzyme, nucleic acid structure, be selected from the polysaccharide of somatomedin, hormone, stimulating factor, antibody, vaccine, interferon, interleukin, plant and animal material.
5. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, is characterized in that: described water soluble drug is selected from: insulin, hirudin, vascular endothelial growth inhibitive factor, neurotrophic factor, cell growth factor, osteogenic growth factor, breast iron transfer albumen, interferon, fluorescin, oxaliplatin, carboplatin, nedaplatin, aclarubicin, doxorubicin, epirubicin, mitoxantrone, bleomycin, Bleomycin A5, mitomycin, Erlotinib, gefitinib, imatinib, Dasatinib, alizapride, azasetron, ondansetron, chlorine phosphoric acid, mesna, Rituximab, ibritumomab tiuxetan, Cetuximab, bevacizumab, Anastrozole, aminoglutethimide, formestane, exemestane, teniposide, etoposide, pentostatin, irinotecan, busulfan, chlormethine, Ka Mosiding, Lomustine, homoharringtonine, asparaginase, pegaspargase, cytosine arabinoside, floxuridine, gemcitabine, AZD2171, alendronate, Rosuvastatin, antithrombase, carmofur, docetaxel, vindesine, vincristine, paclitaxel, BAY 43-9006, decitabine, procarbazine, nimodipine, nifedipine, nitrendipine, felodipine, diclofenac, naproxen, tramadol hydrochloride, morphine, nitroglycerin, clonidine, Ismo 20, ticlopidine, acetazolamide, acetaminophen, aminophylline, amitriptyline, ampicillin, amoxicillin, aspirin, beclamide, caffeine, cimetidine, phenobarbital, Camphora, chloromycetin, Chlophenamin, chlorpromazine hydrochloride, CLOF, cloxacillin, codeine phosphate, diazepam, dextromethorphan, ibuprofen, diphhydramine hydrochloride, doxycycline hydrochloride, eprazinone, fenfluramine, ferrous citrate, ferrous fumarate, ferrous sulfate, scopolamine, pseudoephedrine, berberine, indomethacin, levodopa, lithium carbonate, meclofenoxate hydrochloride, methaqualone, methyl An Feitaiming, acetylspiramycin, nitrofurantoin, nortriptyline, narcotine, hydrochloric acid handkerchief is exerted Lamine, phenacetin, Phenylbutazone, hydrochloric acid benzene good fortune is bright, hydrochloric acid amfetamine alcohol, prednisolone, procainamide, propantheline bromide, Doxaphene, Propranolol, sulphuric acid Kui Nier, thioridazine, zinc gluconate, sulfametomidine, tetracycline, streptomycin, gentamycin, trifluomeprazine, alimemazine, vitamin, nuclear-magnetism contrast agent, isotope.
6. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, it is characterized in that: described matrix material refers to the known phospholipid of pharmacy and lipid materials, be selected from natural phosphatidyl choline, hydrogenated phospholipid, synthetic phospholipid and polyethyleneglycol modified derivant thereof, saturated fatty acid glyceride, steroidal.
7. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, it is characterized in that: in the hydrophilic colloid solution that contains medicine of described preparation process a, can also add 0.1-50 part pharmacy generally acknowledged water solublity antioxidant, stabilizing agent, pH adjusting agent, wherein stabilizing agent is selected from: human serum albumin, tartaric acid, succinic acid, cholic acid, deoxycholic acid, fumaric acid, citric acid, Palmic acid, aminoacid.
8. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, it is characterized in that: in the t-butanol solution I of described preparation process b, can also add 0.1-50 part pharmacy generally acknowledged oil-soluble antioxidant, organic acid and ester thereof, wherein organic acid is selected from: formic acid, acetic acid, tartaric acid, Palmic acid, stearic acid, oleic acid.
9. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, it is characterized in that: the drug-loaded liposome lyophilized powder of preparation injects the known solvent for injection of 1000-50000 part pharmacy before use as required, form the drug-loaded liposome of particle diameter within the scope of 100nm-1000nm.
10. the method for preparing lipidosome of a kind of water soluble medicament-entrapping as claimed in claim 1, it is characterized in that: described liposome is by selecting dissimilar phospholipid or lipid materials combination, make the liposome of long circulating liposomes, pH sensitive liposome body that PEG modifies, responsive to temperature liposome, surface band positive charge, bring into play better in gastrointestinal tract blood circulation targeting drug release effect in positioning release medicine or body.
The method for preparing lipidosome of 11. a kind of water soluble medicament-entrappings as claimed in claim 1, it is characterized in that: described liposome is to use separately or be combined in preparation prescription to apply, by injection, oral, mucosa, skin, wound surface, tract administration, performance treatment, diagnosis, prevention, immunity, clean, the application of sterilization, beauty treatment, health care.
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| CN104840415B (en) * | 2014-02-19 | 2019-03-15 | 香港浸会大学 | Long-acting controlled release lipidosome gel composition containing blood-sugar decreasing active and preparation method thereof |
| CN104027308A (en) * | 2014-06-07 | 2014-09-10 | 刘雷 | bFGF long-circulating liposome, preparation and application thereof |
| EP3265063A4 (en) | 2015-03-03 | 2018-11-07 | Cureport, Inc. | Dual loaded liposomal pharmaceutical formulations |
| CN106924715A (en) * | 2015-12-31 | 2017-07-07 | 深圳翰宇药业股份有限公司 | terlipressin liposome and preparation method thereof |
| KR20190067172A (en) * | 2016-09-09 | 2019-06-14 | 아이리시스, 인크. | Liposome cancer composition |
| US20220087975A1 (en) * | 2019-01-11 | 2022-03-24 | Lipomedix Pharmaceuticals Ltd. | Liposome composition comprising liposomal prodrug of mitomycin c and method of manufacture |
| CN110200919A (en) * | 2019-06-12 | 2019-09-06 | 东南大学 | Prussian blue-Decitabine liposome of one kind and preparation method thereof |
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| CN102552182A (en) * | 2012-02-02 | 2012-07-11 | 鲁翠涛 | Colloidal nucleus liposome lyophilized powder and preparation method thereof |
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