CN102335140A - Disulfiram lipid microsphere preparation for injection for treating tumor and preparation method thereof - Google Patents

Abstract

The invention relates to a disulfiram lipid microsphere preparation for injection for treat tumor and a preparation method thereof, belonging to the field of pharmaceutical preparations. The disulfiram lipid microsphere preparation is prepared from the following components in mass percent: 0.1-4% of disulfiram or derivatives thereof, 5-30% of vegetable oil, 0.6-3% of lecithin, 0-4% of polyethylene glycol phospholipid or derivatives thereof, 0-4% of amino acid block copolymer, 0-0.6% of oleic acid or oleate, 0-1% of antioxidant, 0-0.4% of complexing agent, 1-3% of glycerol and balance of water for injection. According to the invention, due to the introduction of the polyethylene glycol phospholipid or derivatives thereof and the amino acid block copolymer, a fat emulsion can be kept in the blood for a longer time, an additional passive target function on tumor is provided for the medicament, the stability of the fat emulsion preparation is improved, and the effect of disulfiram on treating tumor is enhanced.

Description

Injection disulfiram lipid microsphere preparation of treatment tumor and preparation method thereof

Technical field

The present invention relates to a kind of injection disulfiram lipid microsphere preparation of treating tumor and preparation method thereof, it belongs to field of pharmaceutical preparations.

Background technology

(disulfiram DS), has another name called tetraethylthiuram disulfide, alleviating alcohol addiction sulfur to disulfiram, is synthesized out first in 1881, is used as the catalyst of the vulcanization of rubber in the rubber industry at first; The 1830's, DS was for the first time as medicine, as the treatment of scabies; Subsequently, it is found that disulfiram and copper ion have very strong chelation, can reduce the activity of cupric respiratory enzyme, and the enteral anthelmintic is very responsive to disulfiram, so disulfiram is used as the anthelmintic use again; Two the doctor Hald and the Jacobsen in Copenhagen in 1948 chance on, and drink a small amount of wine after taking the anthelmintic disulfiram, can cause symptoms such as flush, headache, stomachache, perspiration, cardiopalmus, dyspnea immediately.People call disulfiram-like reaction to this symptom that occurs of behind the contact disulfiram, drinking.Just be based on disulfiram-like reaction, disulfiram is using as antialcoholic always since over half a century.The chemical constitution of disulfiram is following:

Prior art mainly discloses the preparation and the effect aspect antibiotic, anti-cataract thereof of disulfiram:

" reparation technology of tetraethylthiuram disulfide " of Superfine Chemical Co Ltd, Leqing City's invention disclosed in Chinese patent CN 1316422A (publication number).It relates to the reparation technology of having invented a kind of tetraethylthiuram disulfide, does not relate to its therapeutical effect and preparation research as medicine.

" the antibiotic therapeutic alliance " of the biological company limited invention of e-disclosed in Chinese patent CN 101939026 A (publication number).It relates to having invented and comprises the imidazoles or derivatives thereof with therapeutic activity and the compositions of disulfiram or derivatives thereof, causes or the combination preparation of the infection that causes as being used to treat by the multi-drug resistant antibacterial, does not relate to the preparation process research of disulfiram.

" a kind of high stable anti-cataract medicine " of Shenyang Pharmaceutical University's invention disclosed in Chinese patent CN 1408418A (publication number).It relates to has invented a kind of (DDC)2Zn (DDC) ₂The hydroxypropyl-beta-cyclodextrin inclusion eye drop of Zn is a kind of high stable anti-cataract medicine, has the characteristics of high biological utilisation, high security and high stability.

Shenyang Pharmaceutical University's invention " inclusion compound of disulfiram as eye drops and preparation method thereof " disclosed in Chinese patent CN 1376463A (publication number).It relates to has invented inclusion compound eye drops that a kind of disulfiram and HP-process and preparation method thereof, can add proppant and process the lyophilizing eye drop; Have good water solubility, stable strong characteristics, can prevent and treat cataract preferably.

Combinatorx, Inc.'s invention " being used to treat the compositions and the method for tumor " disclosed in Chinese patent CN 101217956A (publication number).It relates to the test kit of the method for having invented the multiple compositions that is used to treat the medicine of tumor patient, treatment tumor, the multiple medicine that is used to treat cancer and possibly be used to treat the authentication method of the chemical compound combination of tumor patient; Wherein comprised the combination of alleviating alcohol addiction sulfur (being disulfiram) with the anti-inflammatory agent auranofin.

Zhang Hong etc. have prepared anti-cataract medicine disulfiram Emulsion based on the anti-cataract effect of disulfiram, wherein with Ovum Gallus domesticus Flavus lecithin as emulsifying agent; Adopt ultrasonic method; Investigated the chemical stability of disulfiram in the disulfiram Emulsion, measured different Emulsions and formed influence, drawn the disulfiram degraded and obey first order reaction disulfiram stability; Speed constant and temperature relation meet Arrhenius equation, and wherein better prescription can 4 ℃ be preserved 695 days.

Through patent and literature search, the research of the long-acting disulfiram lipid microsphere preparation that is used to treat tumor of not appearing in the newspapers.

The dissolubility of disulfiram (DS) in water is very poor; The disulfiram that is used for alleviating alcohol addiction is oral solid formulation; Be prone under the gastric acid effect, be degraded into diethyldithiocar bamic acid (DDC) after the oral administration administration, DDC is extremely unstable in gastric acid, can be degraded into Carbon bisulfide and diethylamine rapidly.In blood, DS can form blended disulphide or become DDC by the glutathion reductase system reducing in the erythrocyte with the sulfydryl of endogenous protein very soon, but disulfiram blood drug level still is lower than detectability behind the single-dose.

Summary of the invention

The object of the present invention is to provide a kind of injection disulfiram lipid microsphere preparation of treating tumor, it has the advantages that to improve drug effect, reduce toxic and side effects, persistent.

In order to achieve the above object, the present invention realizes through following technical proposals:

The injection disulfiram lipid microsphere preparation of treatment tumor; It is to be prepared from by said mass percent following component; Disulfiram or its derivant 0.1%-4%, vegetable oil 5%-30%, lecithin 0.6%-3%, Polyethylene Glycol phospholipid or derivatives thereof 0.1-4%, aminoacid block copolymer 0.1-4%, oleic acid or oleate 0-0.6%, antioxidant 0-1%, chelating agent 0-0.4%, glycerol 1%-3%, surplus is a water for injection.

The injection disulfiram lipid microsphere preparation of described treatment tumor; It is to be prepared from by said preferred mass percentage ratio following component; Disulfiram 1-3%, middle chain triglyceride 8-15%, lecithin 1.5-2.5%, Polyethylene Glycol phosphatidase 10 .2-1%, aminoacid block copolymer 0.2-1%, enuatrol 0.05-0.2%, vitamin E 0.2-0.8%, glycerol 2-3%, surplus is a water for injection.

The injection disulfiram lipid microsphere preparation of described treatment tumor; It is to be prepared from by said best in quality percentage ratio following component; Disulfiram 2%, middle chain triglyceride 10%, lecithin 1.8%, Polyethylene Glycol phosphatidase 10 .5%, aminoacid block copolymer 0.5%, enuatrol 0.05-0.21%, vitamin E 0.5%, glycerol 2.25%, surplus is a water for injection.

Above-mentioned vegetable oil is a kind of in the long carbochain triglyceride of injection, middle chain triglyceride, Petiolus Trachycarpi oil, Oleum Helianthi, SEMEN COICIS oil, Semen Maydis oil, Fructrs Hippophae seed oil, Radix Angelicae Sinensis oil, Fructus Zanthoxyli oil, soybean oil, Oleum Bulbus Allii, Oleum Curcumae or the Rhizoma Chuanxiong oil; Described lecithin is a kind of in Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline or the synthetic lecithin; Described Polyethylene Glycol phospholipid or derivatives thereof is a kind of in Polyethylene Glycol-cephalin or derivatives thereof, Polyethylene Glycol-cholesterol or derivatives thereof, Polyethylene Glycol-fatty acid ester or derivatives thereof, Polyethylene Glycol-fatty amine or derivatives thereof, Polyethylene Glycol-aliphatic alcohol or derivatives thereof and the segmental Polyethylene Glycol phospholipid derivative of fat-soluble macromolecule; Described aminoacid block copolymer is a kind of in PEG-PLGA copolymer, glutamic acid-PLGA copolymer, lysine-leucine copolymer, leucine-glutamic acid copolymer, aspartic acid-glutamic acid copolymer, the alanine-glutamine copolymer; Described oleate is potassium oleate or enuatrol; Described antioxidant is L-cysteine, sodium sulfite, sodium sulfite, vitamin E or ascorbic acid; The chelating agent of described control cation concn is selected from ethylenediaminetetraacetic acid (EDTA) or other ionic complexing agents.

Pharmaceutical preparation of the present invention is injection.

Another object of the present invention is to provide a kind of method for preparing of injection disulfiram lipid microsphere preparation of treatment tumor of simple, good stability.

The method for preparing of the injection disulfiram lipid microsphere preparation of treatment tumor; Its preparation process is specific as follows: get the disulfiram, vegetable oil, phospholipid, the Polyethylene Glycol phospholipid or derivatives thereof that meet said mass percent; Under the condition of nitrogen protection; Heating, stirring and dissolving must contain the oil solution of medicine; Other gets the water for injection that meets said mass percent, adds glycerol, enuatrol, aminoacid block copolymer, gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under the shear agitation condition, processes colostrum, and the high pressure homogenizer homogenizing filters, and packing feeds nitrogen, sealing.

The preferred preparation process of the injection disulfiram lipid microsphere preparation of treatment tumor is following: it is to be prepared from by said mass percent following component; Disulfiram 1-3%, vegetable oil 8-15%, lecithin 1.5-2.5%, Polyethylene Glycol phospholipid or derivatives thereof 0.1-1%, aminoacid block copolymer 0.1-1%, oleic acid or oleate 0.05-0.2%, antioxidant 0.2-0.8%, glycerol 2-3%, surplus is a water for injection; The preparation process is specific as follows: get the disulfiram, middle chain triglyceride, lecithin, vitamin E, the Polyethylene Glycol phospholipid that meet said mass percent; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min dissolving evenly obtains containing the oil solution of medicine; Other gets the water for injection that meets said mass percent 85-95% weight, adds the glycerol, enuatrol and the aminoacid block copolymer that meet said mass percent, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that will contain medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml through water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Owing to adopt technique scheme, make the present invention have following advantage and effect:

The applicant through experimental study find disulfiram can targeting in a plurality of signal paths, have positive role at anti-tumor aspect:

Disulfiram (DS) can form blended disulphide with crucial sulfydryl effect in type, the II; Thereby the activity that suppresses type and type; Stop untwisting, transcribe and duplicating of tumor cell dna double chain, and cause the fracture of tumor cell double center chain or single stranded DNA ^[8]

Metalloproteases (MMPs) is the endopeptidase that zinc is contained in the active center, and the transfer of it and tumor cell is closely related; DS through with the zinc chelating in MMPs active center, suppress the activity of MMPs, reduce the expression of tumor cell line MMP-2 and MMP-9, what stop tumor cell invades profit and transfer ^[9] Nuclear factor (NF-κ B) is the factor that and anti-apoptotic and chemotherapy resistance property have substantial connection; DS is through the active expression that suppresses NF-κ B indirectly of CKIs enzyme body; And then existence, propagation and the transfer of inhibition tumor cell; Reduced the expression of the ATP efflux pump of P-glycoprotein (P-gp), many drug resistances associated protein 1 (MRP1) and mitoxantrone drug resistance GAP-associated protein GAP simultaneously; Reduce the chemotherapeutics drug resistance, increase the sensitivity of drug-resistant tumor cell chemotherapeutics ^[10,11]

DS can combine with copper and the zinc in active oxygen (SOD) avtive spot, suppresses ultra oxygen compound dismutase (SOD) activity, increases ROS content in the cell, inducing apoptosis of tumour cell.

Based on this; The disulfiram lipid microsphere injection of the treatment tumor of the present invention's development is to be pharmaceutical carrier with the lipid microsphere; With the novel intravenous injection disulfiram preparation of polyethyleneglycol derivative and the finishing of aminoacid block copolymer, have the drug effect of raising, reduce advantages such as toxic and side effects, persistent.Innovative point of the present invention is described below:

1, with the lipid microsphere is preparing carriers disulfiram injection first, the effect of performance disulfiram treatment tumor.Lipid microsphere (Lipid Microsphere) is stable oil-in-water type (O/W) Emulsion of processing with vegetable oil (main component is a fatty acid triglycercide), phospholipid emulsifier, co-emulsifier, isotonic agent and water for injection; But injection for intravenous; Can be fully by organism metabolism and utilization; Security performance is good, and is non-stimulated to vein, is widely used parenteral nutrition agent in the clinical treatment.The particle diameter of lipid microsphere is little and be evenly distributed, and good stability has certain targeting property; Can solve the preparation difficult problem of oily medicine and fat-soluble medicine as pharmaceutical carrier; Obviously reduce the toxic and side effects of medicine, reduce the zest of medicine, improve drug bioavailability.The anti-cataract medicine disulfiram Emulsion that is different from Zhang Hong etc.; The present invention utilizes the antineoplastic agent disulfiram lipid microsphere injection of high pressure homogenization method preparation; Drug loading is high, sterilization stability is good; And, when improving the external chemical stability of disulfiram, mainly improve disulfiram in stability in blood with water soluble polymer PEG and block polymer decorated microsphere surface.

2, adopt water solublity, the polyethyleneglycol modified lipid microsphere of active, nontoxic, the softish linear polymeric of inanimate object, to prolong the circulation time of disulfiram in blood.Can well solve lipid microsphere pharmaceutical preparation through polyethyleneglycol modified lipid microsphere is prone to from the body circulation by liver, the rapid shortcoming of removing of splenic macrophage; Lipid microsphere pharmaceutical preparation is kept the long period in blood, increased the passive target function of medicine.Relevant Polyethylene Glycol prolongs the mechanism of action of lipid microsphere RT in blood and thinks following three kinds basically:

Polyethylene Glycol forms effective macromolecule isolation strip on the lipid microsphere surface; Its sterically hindered blocking-up or postponed lipoprotein in the blood or the interaction of haemproteins and lipid microsphere, this interaction is considered to cause that phospholipid exchange, lipid microsphere are revealed, the lipid microsphere immobilized artificial membrane destroys or dissolving;

macromolecule isolation strip has cut off the direct interaction of lipid microsphere and cell; Polyethylene Glycol isolation strip is blocked or has been delayed plasma protein in the surperficial absorption of lipid microsphere; Effectively stoped its surperficial opsonic action; Reduce the affinity of mononuclear phagocyte system, thereby reach the effect that prolongs the RT of lipid microsphere in blood lipid microsphere.

3, utilize block polymer decorated lipid microsphere, improved the preparation stability under the environment in vivo, prolong the circulation time of disulfiram.Block copolymer is that various polymerization thing fragment is combined on the polymeric chain through different polymerization methodses and has the copolymer of the essential attribute of each independent segments concurrently.According to the arranging situation of monomeric repetitive on the copolymerization chain, block copolymer can be divided at random according to macromolecular structure, replaces, block and graft copolymer.The charged situation of hydrophilic segment in aqueous solution is different on the copolymerization chain, can be divided into not charged, lotus positive electricity, bear electricity fragment.These characteristics have been given the advantage of block copolymer in drug-loading systems such as polymer vesicle, nanocapsule, millimicro ball:

block copolymer through with the physics or the covalent bonds of drug molecule; Increase the insoluble drug dissolubility, improve drug loading;

aminoacid block copolymer has the charged block copolymer of targeting property

to tumor cell; Repulsive interaction with the erythrocyte surface negative electricity; Delay the destruction of erythrocyte, delay the degraded of medicine the microgranule film.

To sum up; The present invention has made up the used for intravenous injection disulfiram lipid microsphere preparation of treatment tumor,

avoid " first pass effect " of gastric acid degraded and liver through the intravenous administration mode; is wrapped in water-insoluble DS in the stable oil nuclear of phospholipid; Avoid the contacting of endogenic sulfydryl and glutathion reductase in the blood, improve DS blood stability;

utilizes PEG possess hydrophilic property, compliance, sterically hindered characteristics; The PEG phospholipid derivative is modified lipid microsphere; Prolong the body-internal-circulation time of preparation; Reduce with erythrocytic and contact and the phagocytosis of reticuloendothelial system (RES); Increase targeting property, improve physics, the chemistry and biology stability of carrier and medicine.

utilizes charged character of aminoacid block copolymer and sterically hindered effect; Repulsive interaction with the erythrocyte surface negative electricity; Delay the destruction of erythrocyte, delay the degraded of medicine the microgranule film.

The used for intravenous injection disulfiram lipid microsphere preparation of the treatment tumor of the present invention's development; Be be pharmaceutical carrier with the lipid microsphere, with the novel intravenous injection disulfiram preparation of polyethyleneglycol derivative and the finishing of aminoacid block copolymer, have and improve drug effect, reduce advantages such as toxic and side effects, persistent, good stability.The introducing of the present invention through Polyethylene Glycol phospholipid derivative and aminoacid block copolymer solved common fats breast poor stability to the modification on fat milk surface; Be prone to from body-internal-circulation by liver, the rapid shortcoming of removing of splenic macrophage; Fat milk is kept the long period in blood; Increased the passive target function of medicine, improved the stability of fat emulsion formulation, strengthened the effect of disulfiram treatment tumor tumor.

Description of drawings

The degradation curve figure (n=3) of Fig. 1 disulfiram in 50% blood plasma.

The degradation curve figure (n=3) of Fig. 2 disulfiram in 1% whole blood.

Average blood plasma disulfiram concentration-time curve figure behind the disulfiram lipide microsphere injection of Fig. 3 rat difference intravenous injection 22.5mg/kg and the disulfiram solution.

Fig. 4 is the cell growing state comparison diagram before and after the disulfiram lipid microsphere is handled.

The specific embodiment

Below in conjunction with embodiment the present invention is described further.Following examples are merely several specific embodiment of the present invention, but design concept of the present invention is not limited thereto, and allly utilize this design that the present invention is carried out the change of unsubstantiality, all should belong to the behavior of invading protection domain of the present invention.

Method among the following embodiment if no special instructions, is conventional method.

Percentage composition among the following embodiment is the quality percentage composition if no special instructions.

Embodiment 1:

Most preferred disulfiram lipid microsphere preparation of the present invention:

Get disulfiram 20.0g, middle chain triglyceride 100.0g, lecithin 18.0g, vitamin E 5.0g, Polyethylene Glycol phosphatidase 15 .0g; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g, enuatrol 1.0g and aminoacid block copolymer 5.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described lecithin of present embodiment is any one in Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline or the synthetic lecithin.

The described Polyethylene Glycol phospholipid of present embodiment is a kind of in Polyethylene Glycol-cephalin or derivatives thereof, Polyethylene Glycol-cholesterol or derivatives thereof, Polyethylene Glycol-fatty acid ester or derivatives thereof, Polyethylene Glycol-fatty amine or derivatives thereof, Polyethylene Glycol-aliphatic alcohol or derivatives thereof and the segmental Polyethylene Glycol phospholipid derivative of fat-soluble macromolecule.

One, to the investigation of disulfiram content range:

Embodiment 2:

The disulfiram lipid microsphere preparation of minimum disulfiram content

Get disulfiram 1.0g, soybean oil 100.0g, hydrogenated yolk lecithin 18.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described lecithin of present embodiment is any one in Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline or the synthetic lecithin.

Embodiment 3:

The disulfiram lipid microsphere preparation of the highest disulfiram content

Get disulfiram 40.0g, Oleum Bulbus Allii 150.0g, Ovum Gallus domesticus Flavus lecithin 24.0g, vitamin E 3.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 4:

The disulfiram lipid microsphere preparation of optimum disulfiram content

Get disulfiram 20.0g, long carbochain triglyceride 100.0g, soybean lecithin 12.0g, vitamin E 5.5g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 870ml, adds glycerol 24.0g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Two, to the investigation of flux oil amount ranges:

Embodiment 5:

Minimum content vegetable and animals oils, liquid oily medicine or as the disulfiram lipid microsphere preparation of the oil solution preparation of drug solvent:

Get disulfiram 5.0g, Oleum Curcumae 50.0g, hydrogenated soy phosphatidyl choline 10.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 900ml, adds glycerol 22.5g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 6:

High-load vegetable and animals oils, liquid oily medicine or as the disulfiram lipid microsphere preparation of the oil solution preparation of drug solvent:

Get disulfiram 25.0g, Rhizoma Chuanxiong oil 300.0g, Ovum Gallus domesticus Flavus lecithin 18.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 650ml, adds glycerol 25.0g, sodium sulfite 5.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment: 7

Intermediate amounts vegetable and animals oils, liquid oily medicine or as the disulfiram lipid microsphere preparation of the oil solution preparation of drug solvent:

Get disulfiram 15.0g, Radix Angelicae Sinensis oil 200.0g, synthetic lecithin 18.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 750ml, adds glycerol 22.5g, sodium sulfite 5.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Three, to the investigation of main emulsifying agent phospholipid amount ranges

Embodiment 8:

The disulfiram lipid microsphere preparation of minimum proportioning content phospholipid preparation:

Get disulfiram 10.0g, Fructrs Hippophae seed oil 100.0g, hydrogenated yolk lecithin 6.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, ascorbic acid 5.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described lecithin of present embodiment is any one in Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline or the synthetic lecithin.

Embodiment 9:

The disulfiram lipid microsphere preparation of high mixture ratio content phospholipid preparation:

Get disulfiram 20.0g, Semen Maydis oil 100.0g, soybean lecithin 30.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g, L-cysteine 5.0g and potassium oleate 0.5g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and potassium oleate under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 10:

The disulfiram lipid microsphere preparation of middle proportioning content phospholipid preparation:

Get dipropyl dithiocarbamate (disulfiram derivant) 20.0g, Petiolus Trachycarpi oil 100.0g, hydrogenated soy phosphatidyl choline 15.0g, vitamin E 3.0g; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 25.0g, calcium disodium chelate (EDTA-NaCa) 4g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Four, to the investigation of Polyethylene Glycol phospholipid kind and amount ranges

Embodiment 11:

The disulfiram lipid microsphere preparation of mono methoxy polyethylene glycol (MPEG5000) phosphatide complexes:

Get disulfiram 18.0g, soybean oil 150.0g, hydrogenated yolk lecithin 22.0g, vitamin E 5.0g, mono methoxy polyethylene glycol (MPEG5000) phosphatide complexes 2g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g and enuatrol 0.5g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Described PHOSPHATIDYL ETHANOLAMINE-the Macrogol 2000 of present embodiment (PE-PEG2000) can be substituted by in Polyethylene Glycol-cephalin or derivatives thereof, Polyethylene Glycol-cholesterol or derivatives thereof, Polyethylene Glycol-fatty acid ester or derivatives thereof, Polyethylene Glycol-fatty amine or derivatives thereof, Polyethylene Glycol-aliphatic alcohol or derivatives thereof and the segmental Polyethylene Glycol phospholipid derivative of fat-soluble macromolecule any one.

Embodiment 12:

The disulfiram lipid microsphere preparation of cetomacrogol 1000 (PEG1000)-cholesterol:

Get disulfiram 20.0g, Oleum Helianthi 140.0g, Ovum Gallus domesticus Flavus lecithin 20.0g, sodium sulfite 4.0g, cetomacrogol 1000 (PEG1000)-cholesterol 10g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 810ml, adds glycerol 22.5g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 13:

The disulfiram lipid microsphere preparation of low proportioning content Polyethylene Glycol phospholipid:

Get disulfiram 15.0g, SEMEN COICIS oil 100.0g, Ovum Gallus domesticus Flavus lecithin 12.0g, vitamin E 5.0g, DSPE-Macrogol 2000 (DSPE-PEG2000) 1g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described Polyethylene Glycol phospholipid of present embodiment is any one in Polyethylene Glycol-cephalin or derivatives thereof, Polyethylene Glycol-cholesterol or derivatives thereof, Polyethylene Glycol-fatty acid ester or derivatives thereof, Polyethylene Glycol-fatty amine or derivatives thereof, Polyethylene Glycol-aliphatic alcohol or derivatives thereof and the segmental Polyethylene Glycol phospholipid derivative of fat-soluble macromolecule.

Embodiment 14:

The disulfiram lipid microsphere preparation of high mixture ratio content Polyethylene Glycol phospholipid:

Get disulfiram 15.0g, Petiolus Trachycarpi oil 130.0g, Ovum Gallus domesticus Flavus lecithin 12.0g, PHOSPHATIDYL ETHANOLAMINE-Macrogol 2000 (PE-PEG2000) 40g; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 820ml, adds sodium sulfite 4.0g, glycerol 24.0g, disodiumedetate (EDTA-2Na) 2g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 15:

The disulfiram lipid microsphere preparation of middle proportioning content Polyethylene Glycol phospholipid:

Get disulfiram 15.0g, soybean oil 100.0g, Ovum Gallus domesticus Flavus lecithin 16.0g, vitamin E 5.0g, DSPE-Macrogol 2000 (DSPE-PEG2000) 20g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 25.0g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Five, the investigation of aminoacid block copolymerization species and amount ranges:

Embodiment 16:

Polyglutamic acid-gather leucine copolymer (E ₈₀L ₂₀) the disulfiram lipid microsphere preparation modified:

Get disulfiram 10.0g, Oleum Bulbus Allii 150.0g, Ovum Gallus domesticus Flavus lecithin 18.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g, potassium oleate 0.5g and E ₈₀L ₂₀2g, stirring and dissolving is regulated pH5.0-8.0, and dissolving is evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and potassium oleate under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 17:

The disulfiram lipid microsphere preparation that the PEG-PLGA copolymer is modified:

Get disulfiram 10.0g, soybean oil 120.0g, Ovum Gallus domesticus Flavus lecithin 16.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 830ml, adds glycerol 22.5g, enuatrol 1.0g and PEG-PLGA copolymer 1 .5g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described PEG-PLGA copolymer of present embodiment can be by a kind of the substituting in glutamic acid-PLGA copolymer, lysine-leucine copolymer, aspartic acid-glutamic acid copolymer, alanine-glutamine copolymer or the leucine-glutamic acid copolymer.

Embodiment 18:

The disulfiram lipid microsphere preparation of a small amount of aminoacid block copolymer:

Get disulfiram 10.0g, Fructrs Hippophae seed oil 150.0g, Ovum Gallus domesticus Flavus lecithin 15.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 800ml, adds glycerol 22.5g, disodiumedetate (EDTA-2Na) 4g, sodium sulfite 6.0g and aspartic acid-glutamic acid copolymer 2g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

The described aminoacid block copolymer of present embodiment is any one in PEG-PLGA copolymer, glutamic acid-PLGA copolymer, lysine-leucine copolymer or the leucine-glutamic acid copolymer.

Embodiment 19:

The disulfiram lipid microsphere preparation of homoamino acid block copolymer content:

Get disulfiram 10.0g, Oleum Curcumae 200.0g, soybean lecithin 25.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 750ml, adds glycerol 22.5g, ascorbic acid 4.0g and lysine-leucine copolymer 40g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 20:

The disulfiram lipid microsphere preparation of medium proportioning content aminoacid block copolymer:

Get diethyldithiocar bamic acid methyl ester (disulfiram derivant) 30.0g, soybean oil 250.0g, soybean lecithin 25.0gg; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 750ml, adds glycerol 22.5g, enuatrol 2.0g, sodium sulfite 10.0 and alanine-glutamine copolymer 1 8g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Six, to the investigation of co-emulsifier oleic acid or enuatrol or potassium oleate consumption

Embodiment 21:

The disulfiram lipid microsphere preparation that does not add the enuatrol preparation:

Get disulfiram 5.0g, soybean oil 100.0g, Ovum Gallus domesticus Flavus lecithin 12.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, calcium disodium chelate (EDTA-NaCa) 1g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 22:

The disulfiram lipid microsphere preparation of high mixture ratio enuatrol content preparation:

Get disulfiram 5.0g, soybean oil 100.0g, Ovum Gallus domesticus Flavus lecithin 8.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly must contain the oil solution of medicine; Other gets water for injection 800ml, adds glycerol 22.5g and enuatrol or potassium oleate 6.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and oleate; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol or potassium oleate under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Seven, the investigation of antioxidant kind and consumption in the preparation:

Embodiment 23:

The disulfiram lipid microsphere preparation that does not add the antioxidant preparation:

Get disulfiram 20.0g, soybean oil 100.0g, Ovum Gallus domesticus Flavus lecithin 18.0g, Polyethylene Glycol phospholipid (PEG-DSPE) 2.0g; Under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve; Dissolving evenly must contain the oil solution of medicine; Other gets water for injection 850ml, adds glycerol 22.5g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 24:

The disulfiram lipid microsphere preparation of sodium sulfite preparation:

Get disulfiram 15.0g, Fructus Zanthoxyli oil 100.0g, Ovum Gallus domesticus Flavus lecithin 20.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, enuatrol 0.5g and sodium sulfite 2g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 25:

The disulfiram lipid microsphere preparation of L-cysteine preparation:

Get disulfiram 20.0g, soybean oil 100.0g, Ovum Gallus domesticus Flavus lecithin 25.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, enuatrol 1.0g and L-cysteine 1.5g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 26:

The disulfiram lipid microsphere preparation of vitamin E preparation:

Get disulfiram 20.0g, soybean oil 100.0g, soybean lecithin 25.0g and vitamin E 2.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 25.0g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 27:

The disulfiram lipid microsphere preparation of high mixture ratio vitamin E preparation:

Get disulfiram 20.0g, soybean oil 100.0g, soybean lecithin 18.0g, vitamin E 10.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g and enuatrol 0.5g, (EDTA calcium complex disodium salt) EDTA-Ca 2.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Eight, different vegetable and animals oils are as the disulfiram lipid microsphere preparation of flux oil preparation:

Embodiment 28:

Long carbochain triglyceride (12 above fatty acids of carbon atom synthesize with glycerol) is as flux oil:

Get disulfiram 8.0g, long carbochain triglyceride 100.0g, soybean lecithin 15.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, disodiumedetate (EDTA-2Na) 4g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 29:

Middle chain triglyceride (6 ~ 12 carbon atom fatty acid and glycerol synthetic) is as flux oil:

Get disulfiram 16.0g, middle chain triglyceride 100.0g, soybean lecithin 18.0g, vitamin E 4.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 850ml, adds glycerol 22.5g, disodiumedetate (EDTA-2Na) 2g and enuatrol 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 30:

Semen Fructus Hippophae oil is as flux oil:

Get disulfiram 10.0g, Semen Fructus Hippophae oil 140.0g, soybean lecithin 18.0g, vitamin E 5.0g, under the condition of nitrogen protection, be heated to 70 ℃, stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly; Other gets water for injection 810ml, adds glycerol 22.5g and enuatrol 1.0g, disodiumedetate (EDTA-2Na) 1.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 31:

Get disulfiram 10.0g, middle chain triglyceride 100.0g, soybean lecithin 6.0g, vitamin E 8.0g, Polyethylene Glycol phosphatidase 11 0.0g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly must contain the oil solution of medicine; Other gets water for injection 850ml, adds glycerol 25g, enuatrol 2.0g and aminoacid block copolymer 10.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 32:

Get disulfiram 30.0g, middle chain triglyceride 80.0g, soybean lecithin 25.0g, vitamin E 2.0g, Polyethylene Glycol phosphatidase 12 .0g; Under the condition of nitrogen protection; Be heated to 70 ℃; Stir about 10min makes the various lipids that add fully dissolve, and dissolving evenly must contain the oil solution of medicine; Other gets water for injection 870ml, adds glycerol 23g, enuatrol 0.50g and aminoacid block copolymer 2.0g, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and regulates total amount to 1000ml with water for injection; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

Embodiment 33:

Disulfiram lipid microsphere preparation and the agent of disulfiram regular solution are to the inhibitory action of tumor cell

1. material: human breast cancer cell MCF7, disulfiram lipid microsphere preparation (method according to any embodiment of present embodiment 1-32 makes), the agent of disulfiram regular solution

2. method: human breast cancer cell MCF7 cultivated respectively at two kinds contain in the identical culture medium of disulfiram content, wherein a kind of culture medium contains disulfiram lipid microsphere preparation, and another kind contains the disulfiram regular solution, and all the other compositions of culture medium are identical.Breast cancer cell is measured the survival rate of breast cancer cell in the culture medium after cultivating 72 hours under 37 ℃ of conditions.

3. result: contain in the culture medium of disulfiram lipid microsphere preparation, the survival rate of MCF7 cell is 45.61%; Contain in the culture medium of disulfiram regular solution agent, the survival rate of MCF7 cell is 67.35%.

4. conclusion: disulfiram lipid microsphere preparation has the effect of higher inhibition activity of tumor cells with respect to the agent of disulfiram regular solution, has improved the antitumor drug effect of disulfiram.

Embodiment 34:

Disulfiram lipid microsphere preparation is to tumor cell and Normocellular effect:

1. material: human breast cancer cell MCF7, human breast cancer cell MDA-MB-231, normal person's huve cell (HECV), people's normal breast epithelial cell MCF10A, disulfiram lipid microsphere preparation (method according to any embodiment of present embodiment 1-32 makes)

2. method: in the culture medium of four kinds of cells, add the disulfiram lipid microsphere preparation of same dose respectively, cultivate after 1 hour under 37 ℃ of conditions, observation of cell is a growing state.

3. result: as shown in Figure 4, among the figure be before the disulfiram lipid microsphere treated with handle 1 hour after, the growing state of observed different populations of cells in culture medium under 100 power microscopes.After 1 hour, the growth of MCF7 and MDA-MB-231 cell is suppressed through disulfiram lipid microsphere treated, and microscopically does not observe population of cells; And HECV and MCF10A cell normal growth, the visible group of microscopically density does not almost change.

4. conclusion: disulfiram lipid microsphere preparation can distinguish between tumor cells and normal cell, and is little to Normocellular toxicity, can reduce the toxic and side effects in cancer treatment procedure, improves therapeutic index.

 

Nine, the animal experiment study of disulfiram lipide microsphere injection

1, the safety research of disulfiram lipid microsphere

1.1 anaphylaxis experiment

The anaphylaxis that after the administration of observation disulfiram lipide microsphere injection whole body animal is caused.

Dosage divides into groups:

Disulfiram lipide microsphere injection (method according to any embodiment of present embodiment 1-32 makes): priming dose 2ml (40.4mg)/inferior totally three times (i.p.); Challenge dose 2 ml (40.4mg)/inferior be total to once (i.v.)

Disulfiram solution: priming dose 2ml (40mg)/inferior totally three times (i.p.); Challenge dose 2 ml (40mg)/inferior be total to once (i.v.)

The Ovum Gallus domesticus album intravenous fluid: priming dose 2ml/ time is totally three times (i.p.); Challenge dose is total to once (i.v.) 2ml/ time

Negative control group: 5% glucose injection of giving same volume

Get healthy do not have hinder 24 of whole white Cavia porcelluss, be divided into four groups by body weight, sex equilibrium, i.e. disulfiram lipid microsphere injection, disulfiram solution, Ovum Gallus domesticus album intravenous fluid and 5% glucose injection matched group, 6 every group, male and female half and half.I.p. disulfiram lipid microsphere injection, disulfiram solution, Ovum Gallus domesticus album intravenous fluid and 5% glucose injection 2ml next day of every Cavia porcellus difference, continuous three times altogether, the response situation of observing Cavia porcellus after the per injection.I.p. finish, every group is divided into two groups again, three of every groups.One group in the 14th day after being tried thing of injection first; Another organizes the 21st day after injection first; Every Mus is tried thing 2ml respectively accordingly and is attacked more than the i.v.; Have or not after the injection of observed and recorded animal and grab nose, towering hair, dyspnea, spasm, shock, observe 15min continuously until allergic symptoms such as death.Come the overall merit result of the test with anaphylaxis degree, occurrence rate, death condition and Cavia porcellus anaphylaxis progression.

Result of the test: lumbar injection (i.p.) disulfiram lipid microsphere injection next day that healthy albino guinea-pig being distinguished, disulfiram solution, Ovum Gallus domesticus album intravenous fluid, 2ml/ time/Mus of 5% glucose injection, continuous three times.Any abnormal response does not all appear in four groups of Cavia porcelluss after each administration, movable, ingest, drink water all normal, identical with negative control (5% glucose) group.14 days and 21 days difference single intravenous injection (i.v.) disulfiram lipide microsphere injections, disulfiram solution, Ovum Gallus domesticus album intravenous fluid and 5% glucose injection 2ml/ Mus after the first administration.After the injection, disulfiram lipid microsphere injection 2ml (40.4 mg)/inferior/Mus dose groups Cavia porcellus and disulfiram solution 2ml (30mg)/inferior/Mus do not have significant reaction and occur, and anaphylaxis progression is 0.And seriously and significantly symptoms of allergic appears in Ovum Gallus domesticus album group Cavia porcellus, mainly shows as dyspnea, tic, urinary incontinence, promptly shock death occurs, and the death time, all in 2min, mortality rate reached 100%, and anaphylaxis progression is 4 grades.The no abnormal reaction of 5% glucose injection matched group occurs.

The result shows that under this test dose condition, disulfiram lipid microsphere injection is the qualified medicine of hypersensitive test, and the Ovum Gallus domesticus album intravenous fluid has severe anaphylactic reaction to Cavia porcellus.

1.2 external hemolytic experiment

Observe disulfiram lipid microsphere injection and directly contact, whether haemolysis is arranged with blood.

From the about 10ml of rabbit ear edge venous blood collection; Stir 10min except that after defibrinating with Glass rod (top is tied with absorbent cotton); It is centrifugal that (1000rpm * 5min), remove supernatant adds three washings of 10 times of amount normal saline (all centrifugal back abandoning supernatant) again to the supernatant redfree.It is subsequent use that the reuse normal saline is mixed with 2% red blood cell suspension.

Get 8 in 10ml test tube, press table 1 and add various solution.Wherein, the 6th pipe is 5% glucose injection blank, the 7th pipe positive contrast (distilled water).The 8th pipe is the disulfiram solution.After each pipe shakes up gently, place 37 ℃ of water bath with thermostatic control temperature to incubate 4 hours, observe the haemolysis degree of each pipe in 0.5-4 hours, press the standard of table 2 and judge.

The external hemolytic test application of sample of table 1 table

Table 2 haemolysis result judges

Result of the test disulfiram lipid microsphere injection haemolysis or part haemolysis did not all occur at 0.5 hour, 1 hour, 3 hours and 4 hours, showed as that erythrocyte all sinks in the solution, and upper strata liquid water white transparency shakes up the back erythrocyte and disperses; 5% glucose injection group does not all have haemolysis and sticking collection phenomenon yet in 0.5-4 hours, the distilled water group is at the full haemolysis of each time point.See table 3 for details.

The external hemolytic test result of table 3 disulfiram lipid microsphere injection

Can find out that from this test under this experimental condition, disulfiram lipid microsphere injection does not have haemolysis and the sticking collection effect of erythrocyte.

1.3 blood vessel irritation experiment

Observe disulfiram lipid microsphere injection (method according to any embodiment of present embodiment 1-32 makes) continuously after same (concentrated relatively) section blood vessel place, position gives the injection of rabbit auricular vein, to the stimulation of rabbit auricular vein (blood vessel).

First group of disulfiram lipide microsphere injection agent (method according to any embodiment of present embodiment 1-32 makes), 1ml (20.2mg)/only/inferior * 1 time * 1d

Second group of disulfiram solution 1ml (20mg)/only/inferior * 1 time * 1d

The 3rd group of 5% glucose injection 1ml/ only/inferior * 1 time * 1d

Get 9 of rabbit, be divided into three groups by body weight, sex equilibrium, first group of left side auricular vein blood vessel alcohol disinfecting posterior vein injected to disulfiram lipide microsphere injection 1ml (20.2mg)/kg/ time; Second group of left side ear given disulfiram solution 1ml (20mg)/kg/ time; Compare for isometric(al) 5% glucose injection for the 3rd group,, administration 1 time was put to death animal in 24 hours after the administration, 1cm and 5cm place under the inserting needle position, the clip rabbit ear, naked eyes are it is thus clear that have or not redness, mound speckle formation etc.And with 10% formaldehyde fixed, paraffin section, HE dyeing, light microscopic is observed blood vessel endothelium, subcutaneous tissue and thrombosis situation down.

Experimental result shows, disulfiram lipide microsphere injection group rabbit Non Apparent Abnormality in the administration process.And first-class reaction appears screaming, gets rid of in disulfiram solution administration process rabbit.

Finding of naked eye: variation such as disulfiram lipide microsphere injection treated animal liquid medicine injection ear vein blood vessel is not seen obvious congestion, oozed out, edema and necrosis.

Finding under the mirror: disulfiram lipide microsphere injection 1ml (20.2mg)/kg/ group; More or less to include erythrocyte at 1cm place blood vessel under the inserting needle position; Do not see thrombosis in the blood vessel; Vascular endothelial cell has two routine mild swellings, does not see hypertrophy, and that blood vessel is not all seen outward is hemorrhage, edema, necrosis and inflammatory cell infiltration; Do not see that thrombosis, endotheliocyte do not see swelling, hypertrophy under the inserting needle position in the 5cm place blood vessel, blood vessel is not seen the hemorrhage pathological change of Denging outward; All do not see thrombosis in the 1cm place blood vessel under the disulfiram solution injection site, the equal swelling of 3 routine endotheliocytes, hypertrophy in various degree, that blood vessel is not seen outward is hemorrhage, edema and necrosis etc.5cm does not see thrombosis in the place, does not see hemorrhage, edema, necrosis and inflammatory cell infiltration around the blood vessel, but the also visible mild swelling of vascular endothelial cell.5% glucose injection group: 1cm and 5cm place blood vessel and surrounding tissue are not all seen obviously unusual.

Conclusion (of pressure testing): the clinical consumption of disulfiram lipid microsphere injection has slight zest to rabbit auricular vein blood vessel 1cm place under the injection site; And the disulfiram solution all has zest to rabbit auricular vein blood vessel 1cm and 5cm place under the injection site.

2, the degradation kinetics of disulfiram lipid microsphere in blood plasma, whole blood

Getting concentration is the disulfiram lipid microsphere injection 100 μ L of 20.3mg/ml, and the 4mL of 37 ℃ of preheatings of adding contains the normal saline of 50% fresh rat blank plasma and contains in the normal saline of the blank whole blood of 1% fresh rat, hatches for 37 ℃, respectively at 0.17,0.33; 0.45,1,2,4,6; 8,10,12,24 hours sampling 100 μ L, mark liquid (propyl p-hydroxybenzoate methanol solution) 20 μ L in adding; Add chloroform 4mL and extract, centrifugal 10 min of 6000 rpm draw chloroform layer, and nitrogen dries up in 40 ℃ of water-baths, and residue adds methanol 200 μ l and redissolves; Get 20 μ l and measure in HPLC, record disulfiram and interior target peak area calculate both ratio, adopt inner mark method ration.See Fig. 1 and Fig. 2.

The degradation results of table 4 disulfiram in blood

Experiment conclusion: disulfiram lipid microsphere injection can significantly improve disulfiram in stability in blood, prolongs disulfiram RT in vivo, long-acting and tumor locus, performance anti-tumor activity.

3, disulfiram lipide microsphere injection rat plasma pharmacokinetic

3.1 material and method

3.1.1 medicine and reagent

Disulfiram lipide microsphere injection (method according to any embodiment of present embodiment 1-32 makes), specification 10ml:203mg, self-control; The disulfiram solution, specification 10ml:200mg, self-control; Chromatographically pure methanol, chloroform, ether, king Yu chemical plant, Shandong; Propyl p-hydroxybenzoate, rich Dihua, last Tianjin worker's company limited, content 99.41%; Experimental water, double distilled water

3.1.2 animal

Wistar kind rat, body weight (250 ± 30) g, male and female half and half are provided by Shenyang Pharmaceutical University experimental animal center.

3.2 dosage regimen and sample collecting

Body weight is 12 of (250 ± 20) g Wistar male rats, is divided into two groups at random, 6 every group.Fasting 12 hours before the experiment, the dosage of pressing disulfiram 22.5 mg/kg be respectively at femoral vein intravenous injection disulfiram lipide microsphere injection and disulfiram solution, after administration 0.0833,0.25; 0.5,1,1.5,2; 4,6,8,12 and 24 h get about 0.5 ml of blood in the eye socket vein and place the heparinization test tube; Centrifugal 10 min of 6 000 rpm, separated plasma ,-20 ℃ of freezing preservations, to be measured.

3.3 plasma sample is handled

The accurate plasma sample 100 μ l that draw accurately add mark liquid 20 μ l in the propyl p-hydroxybenzoate, shake up, and add 4 ml chloroforms; Vortex extracts 10 min, and centrifugal 10 min of 6000 rpm get chloroform layer; Nitrogen dries up in 40 ℃ of water-baths, and residue adds methanol 200 μ l and redissolves, and gets 20 μ l and measures in HPLC; Record disulfiram and interior target peak area calculate both ratio, adopt inner mark method ration.

3.4 pharmacokinetic experiment result

The blood plasma disulfiram concentration (n=6) of each time point behind the table 5 rat intravenous injection disulfiram lipid microsphere injection

The blood plasma disulfiram concentration (n=6) of each time point behind the table 6 rat intravenous injection disulfiram solution

3.5 average blood drug level-time graph is seen Fig. 3.

Curve shape basically identical when curve is with the medicine that waits after the intravenous injection of dosage disulfiram solution during medicine after the intravenous injection of disulfiram lipide microsphere injection.Can find out by plasma concentration curve; Lower at administration initial stage solution blood drug level than lipid microsphere injection; This is possible because medicine is back by the metabolism of glutathion reductase system reducing in intravenous injection gets into the rat body; The medicine of lipid microsphere parcel is not by the identification of the albumen of glutathion reductase system, so have protective effect.Prolong in time; The disulfiram concentration of lipide microsphere injection is all the time a little more than the solution group; This is because disulfiram is wrapped in the stable oil nuclear of phospholipid and aminoacid block copolymer, avoids the contacting of endogenic sulfydryl and glutathion reductase in the blood, and raising DS blood is stable; Steric hindrance and the charged copolymer and the erythrocytic electrostatic repulsion effect of the PEG chain of microsphere surface simultaneously; Reduce with erythrocytic and contact and the phagocytosis of reticuloendothelial system; Prolonged the body-internal-circulation time of lipid microsphere injection; Improved the chemistry and the biological stability of disulfiram, reached and persist to tumor locus, the purpose of performance anti-tumor activity.

Claims (6)

1. treat the injection disulfiram lipid microsphere preparation of tumor; It is characterized in that it is to be prepared from by said mass percent following component; Disulfiram or its derivant 0.1%-4%, vegetable oil 5%-30%, lecithin 0.6%-3%, Polyethylene Glycol phospholipid or derivatives thereof 0.01-4%, aminoacid block copolymer 0.01-4%, oleic acid or oleate 0-0.6%, antioxidant 0-1%, chelating agent 0-0.4%, glycerol 1%-3%, surplus is a water for injection.

2. the injection disulfiram lipid microsphere preparation of treatment tumor according to claim 1; It is characterized in that it is to be prepared from by said mass percent following component; Disulfiram 1-3%, middle chain triglyceride 8-15%, lecithin 1.5-2.5%, Polyethylene Glycol phosphatidase 10 .2-1%, aminoacid block copolymer 0.2-1%, enuatrol 0.05-0.2%, vitamin E 0.2-0.8%, glycerol 2-3%, surplus is a water for injection.

3. the injection disulfiram lipid microsphere preparation of treatment tumor according to claim 2; It is characterized in that it is to be prepared from by said mass percent following component; Disulfiram 2%, middle chain triglyceride 10%, lecithin 1.8%, Polyethylene Glycol phosphatidase 10 .5%, aminoacid block copolymer 0.5%, enuatrol 0.05-0.21%, vitamin E 0.5%, glycerol 2.25%, surplus is a water for injection.

4. the injection disulfiram lipid microsphere preparation of treatment tumor according to claim 1 is characterized in that described vegetable oil is a kind of in the long carbochain triglyceride of injection, middle chain triglyceride, Petiolus Trachycarpi oil, Oleum Helianthi, SEMEN COICIS oil, Semen Maydis oil, Fructrs Hippophae seed oil, Radix Angelicae Sinensis oil, Fructus Zanthoxyli oil, soybean oil, Oleum Bulbus Allii, Oleum Curcumae or the Rhizoma Chuanxiong oil; Described lecithin is a kind of in Ovum Gallus domesticus Flavus lecithin, soybean lecithin, hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline or the synthetic lecithin; Described Polyethylene Glycol phospholipid or derivatives thereof is a kind of in Polyethylene Glycol-cephalin or derivatives thereof, Polyethylene Glycol-cholesterol or derivatives thereof, Polyethylene Glycol-fatty acid ester or derivatives thereof, Polyethylene Glycol-fatty amine or derivatives thereof, Polyethylene Glycol-aliphatic alcohol or derivatives thereof and the segmental Polyethylene Glycol phospholipid derivative of fat-soluble macromolecule; Described aminoacid block copolymer is a kind of in PEG-PLGA copolymer, glutamic acid-PLGA copolymer, lysine-leucine copolymer, leucine-glutamic acid copolymer, aspartic acid-glutamic acid copolymer, the alanine-glutamine copolymer; Described oleate is potassium oleate or enuatrol; Described antioxidant is L-cysteine, sodium sulfite, sodium sulfite, vitamin E or ascorbic acid; The chelating agent of described control cation concn is selected from EDTA or other ionic complexing agents.

5. the method for preparing of the injection disulfiram lipid microsphere preparation of claim 1,2 or 3 described treatment tumors; It is characterized in that the preparation process is specific as follows: get the disulfiram, vegetable oil, phospholipid, the Polyethylene Glycol phospholipid or derivatives thereof that meet said mass percent; Under the condition of nitrogen protection; Heating, stirring and dissolving must contain the oil solution of medicine; Other gets the water for injection that meets said mass percent, adds glycerol, enuatrol, aminoacid block copolymer, gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that contains medicine adds in the aqueous solution of glycerol and enuatrol under the shear agitation condition, processes colostrum, and the high pressure homogenizer homogenizing filters, and packing feeds nitrogen, sealing.

6. the method for preparing of the injection disulfiram lipid microsphere preparation of the described treatment tumor of claim 5; It is characterized in that it is to be prepared from by said mass percent following component; Disulfiram 1-3%, vegetable oil 8-15%, lecithin 1.5-2.5%, Polyethylene Glycol phospholipid or derivatives thereof 0.1-1%, aminoacid block copolymer 0.1-1%, oleic acid or oleate 0.05-0.2%, antioxidant 0.2-0.8%, glycerol 2-3%, surplus is a water for injection; The preparation process is specific as follows: get the disulfiram that meets said mass percent, middle carbochain triglyceride, lecithin, vitamin E, Polyethylene Glycol phospholipid; Under the condition of nitrogen protection; Be heated to 70 ℃, stir about 10min dissolving evenly obtains containing the oil solution of medicine; Other gets the water for injection that meets said mass percent 85-95% weight, adds the glycerol, enuatrol and the aminoacid block copolymer that meet said mass percent, and stirring and dissolving is regulated pH5.0-8.0, and dissolving evenly gets the aqueous solution of glycerol and enuatrol; Under the condition of nitrogen protection, the oil solution that will contain medicine adds in the aqueous solution of glycerol and enuatrol under shear agitation, processes colostrum, and passes through water for injection residual accommodation total amount to 1000ml; High pressure homogenizer homogenizing 6-15 time, homogenization pressure is approximately 80MPa, to particle size range at 180nm-250nm, filter; Packing feeds nitrogen, sealing, 121 ℃ of rotation sterilization 10min; After lamp inspection was qualified, packing was in storage below 25 ℃.

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