CN102935068A - Preparation method of liposome entrapping water-soluble medicines - Google Patents

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

A kind of method for preparing lipidosome of water soluble medicament-entrapping

[technical field]

The invention belongs to the 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 that is comprised of the orderly lipid bilayer of arrangement.Liposome belongs to colloid system, has the cytoid structure of class, and is strong with the cell membrane affinity, can increase the ability of encapsulated medicine permeate through cell membranes.The liposome good biocompatibility can realize that targeting is sent in the medicine body, have 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 slow-releasing and controlled-releasing action of medicine and the purpose of targeted delivery; (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 realized sterilization or sterile working easily.

The preparation method of liposome has multiple, such 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, the method for preparing lipidosome of existing water soluble medicament-entrapping remains in deficiency, and for example reverse phase evaporation needs long heat treatment process, is unsuitable for the protein and peptide drugs of poor heat stability, and organic solvent is residual easily in the preparation process, and potential hazard is large.Realize easily the asepticize operation in conjunction with thawing law technology freeze-drying method repeatedly, the lipid freeze-dry powder good stability that obtains, but meet water when forming liposome solutions in conjunction with the lipid freeze-dry powder that the freeze-drying method of thawing law technology repeatedly prepares, particle diameter increases several times usually, form is rounding and easily gathering not, can't effectively control particle size range, have safety issue.

Patent of invention " a kind of new method for preparing liposome " (application number 03111470.9) discloses a kind of new method for preparing liposome, its claim 1 is: " a kind of new method for 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 comprised 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 that obtains is obtained liposome ".

Patent of invention " a kind of new method for preparing liposome " (application number 03111470.9) is although utilized the freeze drying process of the tert-butyl alcohol for solution, and clearly emphasize: (1) water-solubility carrier is sucrose, lactose or mannitol; (2) must obtain single phase soln system before the lyophilization.The liposome particle diameter that forms is little although the lipid freeze-dry powder that this patent of invention prepares is dissolved in the water, but structure still is traditional unilamelar liposome, is dissolved in the dissolution medium because the micromolecular water soluble substances such as sucrose, lactose or mannitol see through lipid film easily.Should produce hyperosmosis after the micromolecular water solubleness carrier dissolving of liposome owing to inside of invention preparation in addition, therefore the problem that has the quick seepage of water soluble drug, and this invention entrapment efficiency is lower, and (embodiment 5 is 21%, embodiment 6 is 40%), can't satisfy 2010 editions requirements for liposome encapsulation (greater than 85%) of Chinese Pharmacopoeia.

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, mixing forms 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 in the oil phase forms the w/o type emulsion, ultrasonic, high-speed stirred or high pressure homogenize are processed and are formed the 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 the medicine carrying composite lipidosome: with water soluble drug and amphipathy macromolecule material dissolves in aqueous phase system, carrying out the lyophilization first time processes, lyophilized powder and the Liposome film-forming material that then will comprise water soluble drug are dispersed in the organic facies system, carry out the freeze-drying method second time, make the liposome of water soluble medicament-entrapping.Patent of invention CN201010131244.5 discloses a kind of preparation method of wrapping the medicine carrying composite lipidosome: medicine is scattered in the amphipathy macromolecule material system of dissolving or molten condition, rapid cooling is processed and is obtained medicine solid dispersion, then medicine solid dispersion and Liposome film-forming material are dispersed in the organic facies system, according to method for preparing lipidosome, make the liposome of bag medicine carrying thing.More than three patents of invention, part has solved the problems such as the poor stability that exists among the preparation technology of liposome entrapment medicine or complicated operation, but operating time control is had relatively high expectations or is needed heating and melting processing etc., increase the destruction to macromolecular drug, increased simultaneously the in batches operation control difficulty of preparation.

In a word, the method for preparing lipidosome of existing water soluble medicament-entrapping adopts Emulsion system or double emulsion system mostly, and need through long heat treatment process, because the characteristics such as easy gathering of macromole water soluble drug poor heat resistance, oil-water interfaces, the liposome of the water soluble medicament-entrapping of this class methods preparation of using can not guarantee the activity of the activity, particularly macromolecular drug of medicine.In addition, the liposome of the water soluble medicament-entrapping of this class methods preparation exists that microgranule is easily assembled, the 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 the active drug delivery technologies need to be dialysed, long and poor reproducibility of operating time also is 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 gelatin, collagen or albumin etc. to be contained in the high molecular weight hydrophilic colloid of amino acid structure, form W/W type colloid solution after adding poloxamer (Poloxamer) mixing, lyophilization, the lyophilized powder that forms changes in the t-butanol solution that contains matrix material, again lyophilization after being uniformly dispersed forms the liposome of the water soluble medicament-entrapping of high envelop rate.This method for preparing lipidosome utilizes the secondary freeze drying technology to solve that the method for preparing lipidosome of existing water soluble medicament-entrapping is little for the water soluble drug drug loading, envelop rate is low and the problem of poor stability, and the drug-loaded liposome particle diameter for preparing is in nanoscale, is uniformly dispersed, drug loading is large, entrapment efficiency is high, drug release is slow.The liposome of the applicable preparation of the method water soluble medicament-entrapping, be particularly useful for preparing the liposome of the macromolecular drugs such as the medicine of bag heat-carrying poor stability and albumen, polypeptide, polysaccharide, reach the purpose of slow controlled release drug administration or targeted delivery administration, also can be used for covering bitterness or stink, with other set isolations from, reduce the medicine irritation effect, liquid medicine is converted to the preparation purposes such as solid form, can be applicable to inject, oral, mucosa, skin, wound surface, the part of tract or the multiple dosage form of whole body therapeutic.

The inventor finds in long term test: 1. the high molecular weight hydrophilic colloid solution that contains amino acid structure such as gelatin, collagen or albumin has preferably ability to arrange jobs for water soluble drug, but be difficult to long preservation, can form W/W type colloid solution behind the high molecular weight hydrophilic colloid that contains amino acid structure and poloxamer (Poloxamer) mixing, macromolecular drug is scattered in the interior water of the high molecular weight hydrophilic colloid solution composition that contains amino acid structure easily; 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 made of W/W type colloid solution is scattered in the little and stable height of the liposome particle diameter that forms in the phospholipid solution again.The inventor is through great many of experiments, organically blending is integral with " W/W type colloid solution technology " and " secondary freeze drying technique " innovatively, the characteristics such as 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, overcome the problems such as the drug loading that the traditional liposomal water soluble medicament-entrapping exists is low, entrapment efficiency is low, burst effect is obvious.

The key technology of method for preparing lipidosome of the present invention is combined as: 1. formation W/W type colloid solution behind the preparation high molecular weight hydrophilic colloid that contains amino acid structure and poloxamer (Poloxamer) mixing, and macromolecular drug is scattered in the interior water of the high molecular weight hydrophilic colloid solution that contains amino acid structure easily; 2. adopt freeze drying process that the interior W/W type macromolecule glue liquid solution that holds mutually macromolecular drug is made lyophilized powder, be conducive to keep the high degree of dispersion state of medicine and keep stability; 3. the hydrophilic colloid lyophilized powder that holds water soluble drug changes in the t-butanol solution that contains matrix material, utilize ultrasonic technique to make phospholipid material fully wrap up hydrophilic colloid and be the microgranule of core, form the 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.

Take 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, mixing forms the hydrophilic colloid solution I, and 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 among the t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in the liquid nitrogen, and lyophilization forms the lipid freeze-dry powder that bag carries bovine serum albumin.

The inventor is provided with controlled trial and observes, and the 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 1000 parts of water, mixing forms the hydrophilic colloid solution I, and 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 among the t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in the liquid nitrogen, and lyophilization forms the lipid freeze-dry powder that bag carries bovine serum albumin.

Do not add poloxamer in the controlled trial group 2(hydrophilic colloid): according to Mass Calculation, 1 part of gelatin is dissolved in 1000 parts of water, adds 5 parts of bovine serum albumin, mixing forms the hydrophilic colloid solution I, and 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 among the t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in the liquid nitrogen, and lyophilization forms the lipid freeze-dry powder that bag carries bovine serum albumin.

Controlled trial group 3(only primary freeze drying processes): 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, mixing forms the 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.The hydrophilic colloid solution I is joined among the t-butanol solution I, and ultrasonic (60KHz) processes 1min, is uniformly dispersed, and freezes to form solid in the liquid nitrogen, and lyophilization forms the lipid freeze-dry powder that bag carries 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 mixings and form 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 to formation w/o type emulsion in the oil phase, and 80KHz supersound process 2min forms the 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 the dissolving of 5 parts of bovine serum albumin ,-30 ℃ 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 the oil phase, adds 200 portions of mannitol, ultrasonic (80KHz) processing 2min makes and is 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 ℃ of water-baths, add 5 parts of bovine serum albumin and be uniformly dispersed, go to quenching processing in 0 ℃ 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 the oil phase, adds 200 portions of mannitol, ultrasonic (80KHz) processing 2min makes and is 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, the liposome particle diameter is less, and (mean diameter=810nm), the microscopic pattern rounding is uniformly dispersed, and placing did not have significant change (Fig. 1) in 24 hours.Adopt the 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 the bovine serum albumin liposome does not have burst effect.

Controlled trial group 1(does not add the macromolecule hydrophilic colloid) the bovine serum albumin lipid freeze-dry powder be dissolved in 2000 parts of waters for injection, the liposome initial particle is less, but aggregation velocity is fast, form easily macroparticle (〉 2 μ m), initial envelop rate reaches 86.2%, but drug leakage is serious, has obvious burst effect.

Do not add poloxamer in the controlled trial group 2(hydrophilic colloid) the bovine serum albumin lipid freeze-dry powder be dissolved in 2000 parts of waters for injection large (〉 the 2 μ m of particle diameter), shape is rounding not, and skewness, liposome encapsulation is 89.6%.

Only primary freeze drying processing of controlled trial group 3() bovine serum albumin lipid freeze-dry powder is dissolved in 2000 parts of waters for injection, large (〉 the 2 μ m of liposome particle diameter), shape is rounding not, and liposome encapsulation is 70.2%.

Controlled trial group 4(patent of invention CN201210022488.9 method) in water being distributed to the preparation manipulation that forms the w/o type emulsion in the oil phase, must adopt the supersound process of higher-wattage could form the w/o type microemulsion solution.The bovine serum albumin lipid freeze-dry powder for preparing is dissolved in 2000 parts of waters for injection, and the liposome particle diameter is large, and (mean diameter=980nm), micro-shape be rounding very not, and liposome encapsulation is 88.2%.

Controlled trial group 5(patent of invention CN201010131339.7 method) in the preparation manipulation that solid-state dried frozen aquatic products is distributed in the oil phase, must adopt the supersound process of higher-wattage just can make Solution Dispersion even.The bovine serum albumin lipid freeze-dry powder for preparing is dissolved in 2000 parts of waters for injection, and liposome particle diameter large (mean diameter=1.1 μ m), micro-shape be rounding very not, and liposome encapsulation is 86.1%.

Controlled trial group 6(patent of invention CN201010131244.5 method) in the preparation manipulation that solid dispersion is distributed in the oil phase, must adopt the supersound process of higher-wattage just can make Solution Dispersion even, the bovine serum albumin lipid freeze-dry powder for preparing is dissolved in 2000 parts of waters for injection, the liposome particle size distribution range is wider, mean diameter is large (mean diameter=1.5 μ m), it is irregularly shaped that micro-shape is, and liposome encapsulation is 85.7%.

Experimental result shows that the key technology combination of method for preparing lipidosome of the present invention is indispensable.The contrast prior art, " W/W type colloid solution technology " and " secondary freeze drying technique " combination technology of adoption of innovation of the present invention, guarantee better the activity of high molecular weight protein class medicine, improved the controllability of operation, reduced the difficulty of processing procedure, especially reduced forming the required mechanical treatment intensity (such as ultrasonic time and power) of w/o type microemulsion solution, also improved drug-loaded liposome rounding property and envelop rate.

In addition, the inventor increases by 1 times with the tert-butyl alcohol amount among the poloxamer consumption in the hydrophilic colloid solution I of experimental group and the water yield, the t-butanol solution I, all the other steps are constant, the liposome mean diameter that the bag that obtains carries bovine serum albumin is 550nm, show the adjusting by poloxamer consumption or liquor capacity in the 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; The 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; The bovine serum albumin lipid freeze-dry powder of experimental group preparation 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 according to the clinical practice needs, select suitable phospholipid material, poloxamer model or the injection water yield.

Also find in the experiment, 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 the mass ratio of phospholipid, and medicine of the present invention: the mass ratio of phospholipid can reach 1:1 even lower, 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, realizes easily the asepticize operation.

Thus, a kind of method for preparing lipidosome of water soluble medicament-entrapping, its preparation process comprises following process:

A. calculate with the quality proportioning, 1 part of high molecular weight hydrophilic glue material that contains amino acid structure is added 500-5000 part water formation solution, add the dissolving of 0.5-50 part water soluble drug and 20-500 part poloxamer, form the W/W type hydrophilic colloid solution that contains medicine, lyophilization forms lyophilized powder I;

B. calculate with the quality proportioning, with 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 among the t-butanol solution I, be uniformly dispersed, form t-butanol solution II, freeze to form solid in liquid nitrogen or the refrigerator, lyophilization forms the lipid freeze-dry powder II of water soluble medicament-entrapping, fill seals preservation in cillin bottle, inject 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 that contains amino acid structure comprises gelatin, collagen and human albumin.

Above-mentioned water soluble drug refers to independent application, cooperates the diagnostic reagent that solubilizing agent is used or drug solubility is used greater than biopharmaceutical macromolecular drug, chemicals, Chinese medicine effective extract, medical science or the field of biology of 0.01mg/ml in specific pH value scope.

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, the 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, the 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, the hydrochloric acid handkerchief is exerted the 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, the 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.

Can also add water solublity antioxidant, stabilizing agent, pH adjusting agent that 0.1-50 part pharmacy is generally acknowledged in the hydrophilic colloid solution that contains medicine of above-mentioned preparation process a, wherein stabilizing agent comprises: human serum albumin, tartaric acid, succinic acid, cholic acid, deoxycholic acid, fumaric acid, citric acid, Palmic acid, aminoacid.

Can also add oil-soluble antioxidant, organic acid and ester thereof that 0.1-50 part pharmacy is generally acknowledged among the t-butanol solution I of above-mentioned preparation process b, 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, forms the drug-loaded liposome of particle diameter in the 100nm-1000nm scope.

Above-mentioned liposome is by selecting the combination of dissimilar phospholipid or lipid materials, make the liposome of long circulating liposomes that PEG modifies, pH sensitive liposome body, responsive to temperature liposome, surface band positive charge, bring into play better in the gastrointestinal tract blood circulation targeting drug release effect in positioning release medicine or the body.

Above-mentioned liposome is to use separately or be combined in the preparation prescription to use, 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, have following advantage: organically blend is integral (1) this liposome preparation technology with " W/W colloid technology " and " secondary freeze drying technique " innovatively, the characteristics such as 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 Effective Raise the efficient of traditional liposomal freeze-drying preparation technology, reduce the lyophilizing Operating Complexity.(3) medicine of liposome of the present invention: the mass ratio of phospholipid can reach 1:1 even lower, 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, realizes easily the asepticize operation; (5) can be by regulating the control liposome particle diameters such as hydrophilic colloid proportioning, tert-butyl alcohol consumption; (6) the liposome particle diameter of the present invention's preparation is in nanometer range, and can effectively control particle diameter, be conducive to biomembrane barrier in medicine transdermal, mucosa or the body, applied widely, can be applicable to multiple dosage form, satisfy 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 be used for covering bitterness or stink, with other set isolations from, reduce the medicine irritation effect, liquid medicine is converted to the preparation purposes such as solid form.

[description of drawings]

Fig. 1: the microscopic pattern of bag year bovine serum albumin liposome (mean diameter=810nm)

Fig. 2: water soluble medicament-entrapping liposome preparation flow sketch map

[specific embodiment]

Now further describe the present invention in conjunction with following example.

Embodiment 1: insulin liposome

Present embodiment prepares insulin liposome, and insulin liposome forms such as table 1.

Table 1 insulin liposome component proportion

Insulin liposome method for preparing freeze-dried powder: shown in shown in Fig. 2 flow process, quality proportioning according to above each component, take by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add insulin, poloxamer (Poloxamer), additives I dissolving, formation contains the hydrophilic colloid solution of medicine, and lyophilization forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in 60 ℃ of tert-butyl alcohols, and mixing forms t-butanol solution I.Lyophilized powder I is joined among the t-butanol solution I, and the ultrasonic 2min of 20KHz forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, 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 and 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.The result shows that the consumption of regulating the tert-butyl alcohol and frozen-dried supporting agent can control the 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

Present embodiment prepares growth hormone liposome, and growth hormone liposome forms such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add growth hormone, poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of growth hormone, lyophilization forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and 55 ℃ of heating in water bath form t-butanol solution I.Lyophilized powder I is joined among the t-butanol solution I, and ultrasonic (60KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, 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

Present embodiment utilizes the growth hormone liposome of embodiment 2 preparations to carry out quality evaluation.

Microscopic pattern and granularmetric analysis: get the 100mg lipid freeze-dry powder and add 2ml water formation liposome solutions, drawing respectively a certain amount of liposome turbid liquor also dyes with 1% phosphotungstic acid, place the morphological characteristic of observing liposome under the scanning electron microscope, you measure the liposome particle diameter by special particle size analyzer application library.

Entrapment efficiency determination: get the 1ml liposome solutions, be splined on Sephadex G-50 gel column, take distilled water as eluent, access the eluting part of different volumes, separate and receive free growth hormone eluting part, utilize the 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 sees Table 3, and the growth hormone liposome of visible the present invention's preparation has preferably particle size distribution and microscopic pattern, envelop rate high (all above 90%), and without burst effect.In addition, (surface potential approximately+42mv), (surface potential approximately-16mv) for the faint negative charge of the surface of liposome band of other prescription for the obvious positive charges of growth hormone liposome surface band of prescription 3 preparations.

The quality evaluation of table 3 growth hormone liposome

Annotate: according to Pharmacopoeia of People's Republic of China (2010 editions) regulation, then there is burst effect in the burst size of the microgranules such as liposome, microsphere in beginning 0.5h greater than 40%.

Embodiment 4: the preparation of hydrochloric doxorubicin liposome

Present embodiment prepares hydrochloric doxorubicin liposome, and hydrochloric doxorubicin liposome forms such as table 4.

Table 4 hydrochloric doxorubicin liposome component proportion

Annotate: DPPC is dipalmitoyl phosphatidyl choline; DSPC is the 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add doxorubicin hydrochloride, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, and the ultrasonic 1min of 50KHz forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, and lyophilization 25h forms the lipid freeze-dry powder II that bag carries doxorubicin hydrochloride, and fill seals preservation in cillin bottle.

What prescription 2 and 3 prepared is long circulating liposomes, and what prescription 4 and 5 prepared is temperature sensitive liposome.

Matched group 1--film dispersion method prepares hydrochloric doxorubicin liposome: take by weighing hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in the pear shape bottle with the 20ml ether dissolution, the rotary evaporation film forming, and continue decompression rotary evaporation 15min and eliminate to ether, add 1mg/ml doxorubicin hydrochloride solution 10ml, behind the 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 the hydrochloric doxorubicin liposome lyophilized powder.

Matched group 2--ammonium sulphate gradient prepares the hydrochloric doxorubicin liposome lyophilized powder: take by weighing hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in the pear shape bottle with the 20ml ether dissolution, the ammonium sulfate 10ml that adds 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 gets blank liposome liquid 10ml, remove the salt of the outer aqueous phase of this blank liposome, to wherein adding doxorubicin hydrochloride drug powder 10mg, behind the stirring 2min, leave standstill 20min again, obtain hydrochloric doxorubicin liposome solution, add mannitol 1g, lyophilization obtains the hydrochloric doxorubicin liposome lyophilized powder.

Matched group 3--reverse phase evaporation prepares the hydrochloric doxorubicin liposome lyophilized powder: take by weighing hydrogenation egg yolk lecithin 400mg, cholesterol 100mg, be transferred in the pear shape bottle with the 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 the Evacet lyophilized powder.

Embodiment 5: the quality evaluation of hydrochloric doxorubicin liposome

Present embodiment utilizes the prescription of embodiment 4 and the hydrochloric doxorubicin liposome of matched group preparation to carry out the quality contrast.

Microscopic pattern and granularmetric analysis: with embodiment 3.

Entrapment efficiency determination: get the 100mg lipid freeze-dry powder and add 2ml water formation liposome solutions, be splined on the SephadexG-50 gel column, take distilled water as eluent, access the eluting part of different volumes, separate and receive the liposome eluting part that bag carries doxorubicin hydrochloride, add Triton-100 and destroy hydrochloric doxorubicin liposome, extract amycin and make amycin solution, utilize the HPLC method to detect amycin content (chromatographic condition: Venusil MP C18 post (416mm * 250mm, 5 μ m), mobile phase is acetonitrile-methanol-10mmolL ^-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 the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, in the bag filter of the molecular cut off 3500 of packing into, two ends fasten, and use digestion instrument and measure release according to the method for 2010 editions pharmacopeia regulations.Dissolution medium is the alcoholic solution 800ml of 0.25% sodium lauryl sulphate and 10%, temperature is 37 ± 0.5 ℃, and mixing speed is 100r/min, timing sampling 5ml, and in time replenishing isothermal equal-volume blank medium, HPLC measures peak area behind the sample 0.22 μ m filtering with microporous membrane.Each time point hydrochloric doxorubicin liposome Cumulative release amount of calculation sample.Prescription 4 and 5 is the responsive to temperature liposome, and the drug release behavior assay method is the same, but the dissolution medium Temperature Setting is 37 ℃, 40 ℃ and 43 ℃ of three levels, and to write out a prescription 1 as contrast.

Experimental result sees 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, the 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 the responsive to temperature liposome.The drug release experiment shows, prescription 1,2,3,6 and 7 contrast groups have preferably slow releasing function, release time is greater than 8h, prescription 4 and 5 responsive to temperature liposome in the time of 37 ℃ 24h cumulative release amount between 80-100%, rate of release is obviously accelerated when temperature is brought up to 40 ℃ or 43 ℃, burst size is near 100% during 43 ℃ of 4h, therefore write out a prescription 4 and the prescription 5 responsive to temperature liposomees that prepare have 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 just will be wrapped the medicine amycin that carries and be changed to topotecan hydrochloride with the prescription 2 of embodiment 4.

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%.The drug release experiment shows that the topotecan hydrochloride liposome has the similar drug release behavior of Evacet with embodiment 5 prescriptions 2, has preferably slow releasing function.

Embodiment 7: the ganoderan liposome

Present embodiment prepares the ganoderan liposome, and the ganoderan liposome forms such as table 6.

The lipidosome freeze-dried powder, preparation method thereof of ganoderan: according to the quality proportioning of above each component, take by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add ganoderan, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of ganoderan, lyophilization forms lyophilized powder I.Matrix material, polyoxyethylene hydrogenated Oleum Ricini, additives II are dissolved in the tert-butyl alcohol, and 50 ℃ of heating in water bath form t-butanol solution I.Lyophilized powder I is joined among the t-butanol solution I, and ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, and lyophilization 25h forms the lipid freeze-dry powder II that bag carries ganoderan, and fill seals preservation in cillin bottle.

Microscopic pattern and granularmetric analysis: method is with embodiment 3.

Entrapment efficiency determination: get the 100mg lipid freeze-dry powder and add 1ml water formation liposome solutions, be splined on the SephadexG-50 gel column, take 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.

The result: the ganoderan liposome of three formula preparations has preferably particle diameter, and mean diameter is less than 1000nm, and without clustering phenomena, the envelop rate of the ganoderan liposome of three formula preparations is all above 90%.

Table 6 ganoderan liposome component proportion

Embodiment 8:bFGF liposome

Present embodiment prepares basic fibroblast growth factor (bFGF) liposome, and the bFGF liposome forms such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add bFGF, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, and ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, and lyophilization 25h forms the lipid freeze-dry powder II that bag carries bFGF, and fill seals preservation in cillin bottle.

Microscopic pattern and granularmetric analysis: get the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the bFGF liposome of each formula preparation has preferably particle diameter, and mean diameter is less than 950nm, without clustering phenomena.In addition, (surface potential approximately+32mv), (surface potential approximately-15mv) for the faint negative charge of the surface of liposome band of other prescription for the obvious positive charges of bFGF surface of liposome band of prescription 8 preparations.

Embodiment 9: the recombinant human vascular endothelial inhibin liposome

Present embodiment prepares the recombinant human vascular endothelial inhibin liposome, and the recombinant human vascular endothelial inhibin liposome forms together such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add recombinant human vascular endothelial inhibin, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, lyophilization 25h forms the lipid freeze-dry powder II of bag load-carrying group human Endostatin, and fill seals preservation in cillin bottle.

Microscopic pattern and granularmetric analysis: get the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the recombinant human vascular endothelial inhibin liposome of each formula preparation has preferably particle diameter, and mean diameter is less than 900nm, without clustering phenomena.

Embodiment 10: transforming growth factor β (TGF-β) liposome

Present embodiment prepares transforming growth factor β (TGF-β) liposome, and TGF-β liposome forms such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add TGF-β, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, and ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, and lyophilization 25h forms the lipid freeze-dry powder II that bag carries TGF-β, and fill seals preservation in cillin bottle.

Microscopic pattern and granularmetric analysis: get the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the TGF-β liposome of each formula preparation has preferably 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: the human interferon-alpha-2 b liposome

Present embodiment prepares the human interferon-alpha-2 b liposome, and the human interferon-alpha-2 b liposome forms such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add human interferon-alpha-2 b, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, and ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, 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 the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the human interferon-alpha-2 b liposome of each formula preparation has preferably 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

Present embodiment prepares the pseudoephedrine liposome take the Effective Component of Chinese Medicine pseudoephedrine as object, and preparation forms such 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 by weighing respectively component, the high molecular weight hydrophilic colloidal materials is added to the water formation solution, add pseudoephedrine, Poloxamer, additives I dissolving, formation contains the hydrophilic colloid solution of 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 among the t-butanol solution I, and ultrasonic (50KHz) 1min forms t-butanol solution II, and quick-freezing forms solid in the liquid nitrogen, and lyophilization 25h forms the lipid freeze-dry powder II that bag carries pseudoephedrine, and fill seals preservation in cillin bottle.

Microscopic pattern and granularmetric analysis: get the 200mg lipid freeze-dry powder and add 2ml water formation liposome solutions, microscopic pattern and granularmetric analysis method are with embodiment 3, the pseudoephedrine liposome of each formula preparation has preferably 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 exemplary description, but those skilled in the art can carry out various modifications to the present invention after reading present patent application in the situation that does not break away from the spirit and scope of the present invention.

Claims (11)

1. the method for preparing lipidosome of a water soluble medicament-entrapping, it is characterized in that: its preparation process comprises following process:

A. calculate with the quality proportioning, 1 part of high molecular weight hydrophilic glue material that contains amino acid structure is added 500-5000 part water formation solution, add the dissolving of 0.5-50 part water soluble drug and 20-500 part poloxamer, form the W/W type hydrophilic colloid solution that contains medicine, lyophilization forms lyophilized powder I;

B. calculate with the quality proportioning, with 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 among the t-butanol solution I, be uniformly dispersed, form t-butanol solution II, freeze to form solid in liquid nitrogen or the refrigerator, lyophilization forms the lipid freeze-dry powder II of water soluble medicament-entrapping, fill seals preservation in cillin bottle, inject 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, it is characterized in that: the described high molecular weight hydrophilic glue material that contains amino acid structure comprises 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, cooperates the diagnostic reagent that solubilizing agent is used or drug solubility is used greater than biopharmaceutical macromolecular drug, chemicals, Chinese medicine effective extract, medical science or the field of biology of 0.01mg/ml in specific pH value scope.

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, comprises 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, it is characterized in that: described water soluble drug comprises: insulin, hirudin, the 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, the 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, the hydrochloric acid handkerchief is exerted the 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, the 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, comprises 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: can also add water solublity antioxidant, stabilizing agent, pH adjusting agent that 0.1-50 part pharmacy is generally acknowledged in the hydrophilic colloid solution that contains medicine of described preparation process a, wherein stabilizing agent comprises: 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: can also add oil-soluble antioxidant, organic acid and ester thereof that 0.1-50 part pharmacy is generally acknowledged among the t-butanol solution I of described preparation process b, wherein organic acid comprises: 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, forms the drug-loaded liposome of particle diameter in the 100nm-1000nm scope.

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 the combination of dissimilar phospholipid or lipid materials, make the liposome of long circulating liposomes that PEG modifies, pH sensitive liposome body, responsive to temperature liposome, surface band positive charge, bring into play better in the gastrointestinal tract blood circulation targeting drug release effect in positioning release medicine or the body.

11. 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 to use separately or be combined in the preparation prescription to use, 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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