CN102319214B - Lipid microsphere injection containing sodium demethyl cantharidate-phosphatide complex and preparation method thereof - Google Patents
Lipid microsphere injection containing sodium demethyl cantharidate-phosphatide complex and preparation method thereof Download PDFInfo
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- CN102319214B CN102319214B CN 201110267058 CN201110267058A CN102319214B CN 102319214 B CN102319214 B CN 102319214B CN 201110267058 CN201110267058 CN 201110267058 CN 201110267058 A CN201110267058 A CN 201110267058A CN 102319214 B CN102319214 B CN 102319214B
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- injection
- demethylate
- water
- phosphatide
- preparation
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a lipid microsphere injection containing sodium demethyl cantharidate-phosphatide complex and preparation method thereof. The injection comprises the sodium demethyl cantharidate-phosphatide complex, fat-soluble medium, surfactant and other ingredients. In the sodium demethyl cantharidate-phosphatide complex, the molar ratio of sodium demethyl cantharidate to phosphatide is 1-1:10. The prepared sodium demethyl cantharidate-phosphatide complex improves the distribution of sodium demethyl cantharidate in oil-water 2-phase interfacial film and oil phase greatly, increases the entrapment efficiency of the medicine in preparation, realizes the interfacial film loading medicine, reduces the toxicity, and can carry out targeting drug release in vivo. The prepared lipid microsphere injection reduces the vascular stimulation of sodium demethyl cantharidate, improves the curative effect, and reduces toxic and side effect.
Description
Technical field
The present invention relates to a kind of lipide microsphere injection and preparation method thereof, be specifically related to a kind of demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid and preparation method thereof that contains, belong to medical technical field.
Background technology
PTS cantharidin (Norcantharidin, NCTD) is removed 1,2 methyl and is got by cantharidin.Demethylate disodium cantharidinate is the sodium salt that the norcantharidin direct hydrolysis makes, and it only dissolves in minority organic solvent (such as acetone, ethyl acetate) and hot water, and the dissolubility in cold water and oil is very low.Compare with cantharidin, the urinary system stimulation of demethylate disodium cantharidinate reduces greatly, has simultaneously than the more significant function of increasing leukocyte of cantharidin.Demethylate disodium cantharidinate is cell cycle specific agents, blocks the cycle in M, and affects its periodic duty speed.Be mainly used in clinically premedicate or the combined chemotherapies such as hepatocarcinoma, esophageal carcinoma, gastric cancer, carcinoma of gastric cardia, can be used for leucocytes reduction and hepatitis B.
At present, demethylate disodium cantharidinate mainly is applied to clinical with tablet and demethylate disodium cantharidinate injection.Although demethylate disodium cantharidinate has obviously alleviated the toxicity of cantharidin, do not eliminate fully.Results of Animal shows that when dosage increased to a certain degree, pathological change appearred in kidney and liver organization.The strict restriction of using dosage has also seriously hindered the performance of curative effect of medication.Untoward reaction appears in some patients were during clinical use when intravenously administrable every day surpasses 20mg or surpasses 30mg oral every day, oral untoward reaction main manifestations for feel sick, vomiting, the symptom such as dizzy.Because injection is alkaline solution, its pH value is about 8, and is therefore larger to the blood vessel wall stimulation much larger than pH value of blood (7.35～7.45), easily causes chemical phlebitis; And because medicine itself has than strong and stimulating, also can cause vasospasm, blood reduces, and relatively local disodium cantharidinate concentration increases, and increases the weight of phlebitis, and if transfusion speed greater than velocity of blood flow, then phlebitic incidence rate obviously increases.Main adverse reaction shows as and easily causes inflammation of vein, and vein section is red, swollen, hot, bitterly, and is rubescent along vein traveling strip, then red and swollenly becomes block if any oozing out, and limitation of activity is processed to take a turn for the better through hot packing and cured.Therefore clinical during for the periphery superficial intravenous infusion, the ratio that superficial phlebitis occurs is quite high, and the patient who has refuses with this medicine treatment, affects clinically normally use of medicine.
Given this, bring into play better clinical efficacy for making demethylate disodium cantharidinate, the research and development novel form, advantage efficient, low toxicity with novel form is given full play to the demethylate disodium cantharidinate anti-tumor activity and is reduced toxic and side effects, and this undoubtedly will be significant to further promotion and the application of popularization cantharidin in clinical.
Summary of the invention
Technical problem to be solved by this invention is that the defective that overcomes prior art provides a kind of demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid, and this demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid has the characteristics such as low blood vessel irritation and hypotoxicity.Further, the invention provides this demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid and get preparation method.
Technical problem of the present invention is realized by following technical scheme.
A kind of demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid, this injection contains demethylate disodium cantharidinate phosphatide complexes, fat-soluble medium, surfactant and water.
Described Injectio natarii norcantharidatis phosphatide complexes is composited in organic solvent by demethylate disodium cantharidinate and phospholipid.The mol ratio of described demethylate disodium cantharidinate and phospholipid is 1～1:10.Described phospholipid is selected from one or more in natural phospholipid or the synthetic phospholipid.Described natural phospholipid is lecithin, fabaceous lecithin, cholesterol, cholic acid class or chitosan; Described synthetic phospholipid is phosphatidyl glycerol ester or distearoyl phosphatidylcholine.Described demethylate disodium cantharidinate is cantharidin or the demethylate disodium cantharidinate that is reacted generation by cantharidin process in the preparation preparation process.Described organic solvent is ethyl acetate, oxolane, dichloromethane, chloroform, acetone, dimethyl sulfoxide, normal hexane, cyclohexane extraction, methanol, ethanol, isopropyl alcohol, n-butyl alcohol.
Described fat-soluble medium comprises oil phase and oil soluble emulsifying agent; Described oil phase comprises mineral oil, vegetable oil, animal oil or its mixture; Described oil soluble emulsifying agent is one or more that are selected from natural material or the synthetic material; Described oil phase is MCT Oil, soybean oil, safflower oil, Semen Maydis oil or its mixture preferably; Described natural material preferably is lecithin, fabaceous lecithin, cholesterol, oleic-acid, cholic acid class or chitosan; Described synthetic material preferably is phosphatidyl glycerol ester, distearoyl phosphatidylcholine.
Described surfactant is selected from phospholipid, tween, poloxamer, enuatrol, oleic acid, cholic acid, deoxycholic acid or its mixture.Described phospholipid is selected from lecithin, fabaceous lecithin or its mixture; Described tween is selected from polysorbas20, polysorbate40, polysorbate60, Tween 80, polysorbate85, or the arbitrary proportion mixture of above different size; Described poloxamer is PLURONICS F87.
Described demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid can also contain isoosmotic adjusting agent, pH adjusting agent and metal-chelator.
Described isoosmotic adjusting agent is glycerol, sorbitol, mannitol, glucose or its mixture; Described pH adjusting agent is sodium hydroxide, hydrochloric acid or buffer salt or its mixture; Described metal-chelator is disodium edetate, sodium calcium edetate or its mixture.
Demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid of the present invention, preferred group becomes by weight percentage:
Injectio natarii norcantharidatis phosphatide complexes (in cantharidin) 0.01%～5%;
Fat-soluble medium 5%～30%;
Surfactant 0.5%～5%;
Isoosmotic adjusting agent 0.5%～5%;
PH adjusting agent 0.01%～1%;
Metal-chelator 0%～1%;
All the other are water for injection.
More preferably:
Demethylate disodium cantharidinate complex (in cantharidin) 0.05 %～0.4%;
Fat-soluble medium 10%～20%;
Surfactant 0.5%～5%;
PH adjusting agent 0.01%～1%;
Metal-chelator 0.005%～0.03%;
All the other are water for injection.
Most preferably be:
Demethylate disodium cantharidinate complex (in cantharidin) 0. 2%;
Fat-soluble medium 10%;
Lecithin 1.2%;
Glycerol 2.5%;
Enuatrol 0.03%;
PLURONICS F87 0.2%;
Disodium edetate 0.02%;
All the other are water for injection;
Wherein said fat-soluble medium is MCT Oil (MCT), soybean oil, safflower oil, Semen Maydis oil or its mixture.
The invention provides a kind of preparation method of demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid, it is characterized in that comprising the steps:
(1) preparation of demethylate disodium cantharidinate phosphatide complexes: remove methyl disodium cantharidinate and phospholipid, it is added in the organic solvent, 25～70 ℃ of reflux 0.5～3 hour are to the solution clear; Take out solution and reduce pressure rotary evaporation to remove organic solvent, place at last vacuum drier or freezer dryer to carry out drying;
(2) with the demethylate disodium cantharidinate phosphatide complexes of recipe quantity, fat-soluble medium heated and stirred under 60～80 ℃ condition, until phosphatide complexes all is dissolved in the fat-soluble medium, get oil phase;
(3) surfactant and the isoosmotic adjusting agent with recipe quantity adds the extremely whole dissolvings of an amount of water for injection heated and stirred under 60～80 ℃ condition as water;
(4) utilize tissue mashing machine, slowly water is added oil phase or oil phase is added aqueous phase, with per minute 10,000～20,000 turns stirring 3～5 minutes, namely gets slightly breast;
(5) thick breast is transferred in the high pressure dispersing emulsification machine, at 600～1200bar, homogenize is 3～10 times under 30 ℃～60 ℃ conditions;
(6) get above-mentioned Emulsion embedding in infusion bottle, fill nitrogen, 115 ℃ of lower sterilization 30min or 121 ℃ of sterilization 15min.
Preferred preparation method is:
(1) preparation of demethylate disodium cantharidinate phosphatide complexes: remove methyl disodium cantharidinate 2g, soybean phospholipid 6g, add ethyl acetate 100ml, 60 ℃ after compound 1 hour, rotary evaporation is gone out ethyl acetate, more than 12 hours (20-30 ℃), namely gets medicine carrying soybean phospholipid complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved;
(2) with demethylate disodium cantharidinate phosphatide complexes and the soybean oil of recipe quantity: MCT=3:1, heated and stirred under 70 ℃ condition is until phosphatide complexes all is dissolved in the oil phase;
(3) glycerol, enuatrol, Tween 80 and the disodium edetate with recipe quantity adds the extremely whole dissolvings of an amount of water for injection heated and stirred under 70 ℃ condition as water;
(4) oil phase is added to aqueous phase, places high-speed tissue mashing machine 1,0000～2,0000rmp stirred 3-5 minute;
(5) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 6.0～8.0, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, with 600～1000 bar pressure homogenizing 6～10 times;
(6) inflated with nitrogen, embedding.Adopt 121 ℃ of sterilizations of high-pressure rotary 15 minutes.
Methyl disodium cantharidinate phosphatide composite liposome microsphere injection liquid of the present invention is the oil-in-water submicronized emulsion that contains demethylate disodium cantharidinate that a kind of intravenous transfusion is used, and the compositions that consists of the oil-in-water submicronized emulsion is identical with the compositions of lipide microsphere injection.
Distribute in the biphase interfacial film of oil phase or profit should be more than 80% for demethylate disodium cantharidinate in the demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid of the present invention, and the remainder demethylate disodium cantharidinate is distributed in water and the oil phase.
Demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid of the present invention is at first compound with demethylate disodium cantharidinate and phospholipid, to in oil, water, be written in the phospholipid layer by the medicine demethylate disodium cantharidinate of equal indissoluble, demethylate disodium cantharidinate improves greatly at the biphase interfacial film of profit and the distribution in the oil phase, improved the envelop rate of medicine in preparation, realize the interfacial film medicine carrying, improved that it is fat-soluble, improved the drug loading of medicine in preparation, reduced its toxicity and made it in vivo can targeting drug release.Then utilize phosphatide complexes to prepare Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid, reduced the blood vessel irritation of demethylate disodium cantharidinate, improve curative effect and reduce toxic and side effects, thereby have certain novelty and stronger practicality.Experimentation shows: the Injectio natarii norcantharidatis lipide microsphere injection is compared with injection, and acute toxicity obviously reduces.After rat vein gives Injectio natarii norcantharidatis lipide microsphere injection and its solution-type injection, the blood medicine time graph of Injectio natarii norcantharidatis is seen Fig. 1, and the drug distribution figure of Injectio natarii norcantharidatis in each tissue of heart, liver, kidney and different time sees Fig. 2-Fig. 7.
Description of drawings
Fig. 1 NCTD blood medicine-time graph 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection
Fig. 2 NCTD heart Chinese medicine concentration-time graph 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection
Fig. 3 NCTD liver Chinese medicine concentration-time graph 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection
Fig. 4 NCTD kidney Chinese medicine concentration-time graph 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection
Respectively organize medicine scattergram 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection in 0.5 hour the rat body of Fig. 5 administration
Respectively organize medicine scattergram 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection in 6 hours the rat body of Fig. 6 administration
Respectively organize AUC value 1 Injectio natarii norcantharidatis lipide microsphere injection 2 Injectio natarii norcantharidatis solution-type injection behind Fig. 7 administration 24 h in the rat body.
The specific embodiment
The invention will be further described below in conjunction with the specific embodiment, and example is only limited protection scope of the present invention for indicative explaination being provided, not meaning that by any way.
The preparation of embodiment 1 demethylate disodium cantharidinate raw material
Although demethylate disodium cantharidinate has injection, all be to make sodium-salt aqueous solution from the norcantharidin direct hydrolysis.Its crude drug does not have production and selling.A organic reaction the most basic and prepare Injectio natarii norcantharidatis from norcantharidin, therefore, our reference literature report, take norcantharidin as raw material, through hydrolysis salifying, dehydration, washing, drying makes Injectio natarii norcantharidatis.Reaction equation is as follows:
Synthetic method is as follows:
Sodium hydroxide 10-30g adds water 100mL, lets cool after the dissolving, adds while stirring norcantharidin 10-30g, after nearly all dissolvings, and a small amount of insoluble matter of filtering.With concentrated near the doing of solution decompression, get a large amount of white powder solids, add ethanol 5-50mL, sucking filtration, obtain white crystal, repeatedly wash with ethanol, wash the pH value of rear inspection eluate at every turn, when treating that the eluate pH value is 4-9, sucking filtration, vacuum drying gets white crystalline powder.
Refining: with above-mentioned white powder 5-50g, be dissolved in the 5-50mL water, filter with the G3 sintered filter funnel, filtrate adds 100-1000mL ethanol, places sucking filtration after 2-20 hour, and vacuum drying gets the white powder solid.
The preparation of embodiment 2 phosphatide complexes
[preparation 1] demethylate disodium cantharidinate-Ovum Gallus domesticus Flavus lecithin complex
Remove methyl disodium cantharidinate (in norcantharidin) 4g, Ovum Gallus domesticus Flavus lecithin 6g, add acetone 50ml, 60 ℃ after compound 1 hour, rotary evaporation is gone out acetone, more than 12 hours (20-30 ℃), namely gets medicine carrying Ovum Gallus domesticus Flavus lecithin complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
[preparation 2] demethylate disodium cantharidinate-soybean lecithin complex
Remove methyl disodium cantharidinate (in norcantharidin) 2g, soybean phospholipid 6g, add ethyl acetate 100ml, 60 ℃ after compound 1 hour, rotary evaporation is gone out ethyl acetate, more than 12 hours (20-30 ℃), namely gets medicine carrying soybean phospholipid complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
[preparation 3] demethylate disodium cantharidinate-cholesterin complex
Remove methyl disodium cantharidinate (in norcantharidin) 1g, cholesterol 3g, add ethyl acetate 100 ml, 60 ℃ after compound 3 hours, rotary evaporation is gone out ethyl acetate, more than 12 hours (20-30 ℃), namely gets the medicine carrying cholesterin complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
[preparation 4] demethylate disodium cantharidinate-phosphatidyl glycerol ester complexes
Remove methyl disodium cantharidinate (in norcantharidin) 0.5g, phosphatidyl glycerol ester 2g, add acetone 50 ml, 60 ℃ after compound 1 hour, rotary evaporation is gone out acetone, more than 12 hours (20-30 ℃), namely gets medicine carrying phosphatidyl glycerol ester complexes through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
[preparation 5] demethylate disodium cantharidinate-distearoyl phosphatidylcholine complex
Remove methyl disodium cantharidinate (in norcantharidin) 0.5g, distearoyl phosphatidylcholine 2g, add ethyl acetate 100 ml, 60 ℃ after compound 1 hour, rotary evaporation goes out ethyl acetate, more than 12 hours (20-30 ℃), namely gets medicine carrying distearoyl phosphatidylcholine complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
[preparation 6] demethylate disodium cantharidinate-two Semen Myristicae phosphatidyl choline complex
Remove methyl disodium cantharidinate 2g, two Semen Myristicae phosphatidyl choline 6g, add oxolane 250 ml, 60 ℃ after compound 1 hour, rotary evaporation goes out oxolane, more than 12 hours (20-30 ℃), namely gets medicine carrying two Semen Myristicae phosphatidyl choline complex through vacuum drying, airtight package is put into refrigerator cold-storage and is preserved.
Embodiment 3 preparation demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquids
[prescription 1] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05 |
| Soybean oil | |
| 10% | |
| Lecithin | 1.2% |
| Glycerol | 2.5% |
| Enuatrol | 0.03% |
| PLURONICS F87 | 0.2% |
| Calcium disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, PLURONICS F87, enuatrol and calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 4] are joined in the injection soybean oil, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 2] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05 % |
| Oil phase (soybean oil: MCT=3:1) | 10% |
| Glycerol | 2.5% |
| Sodium deoxycholate | 0.03% |
| PLURONICS F87 | 0.2% |
| Calcium disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, PLURONICS F87, sodium deoxycholate, calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) [preparation 2] middle phosphatide complexes is joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 3] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 1mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.1% |
| Oil phase (soybean oil: MCT=3:1) | 15% |
| Fabaceous lecithin | 2.5% |
| Glycerol | 2.5% |
| Enuatrol | 0.06% |
| PLURONICS F87 | 0.6% |
| Sodium calcium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, PLURONICS F87, enuatrol, sodium calcium edetate are scattered in an amount of water for injection, are heated to 80 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection fabaceous lecithin in [preparation 3] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.0, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 115 ℃ of sterilization 30 min, the rapid cooling of ice-water bath and get final product.
[prescription 4] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 1mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.1% |
| Oil phase (soybean oil: MCT=1:1) | 20% |
| Lecithin | 3% |
| Mannitol | 2.5% |
| Enuatrol | 0.1% |
| PLURONICS F87 | 0.4% |
| Disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) injection mannitol, PLURONICS F87, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 5] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 5] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 1mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.1% |
| Oil phase (soybean oil: MCT=1:1) | 15% |
| Lecithin | 1.8% |
| Mannitol | 2.5% |
| Enuatrol | 0.06% |
| PLURONICS F87 | 0.4% |
| Disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) injection mannitol, PLURONICS F87, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 1] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 6] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 2mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.2% |
| Oil phase (soybean oil: MCT=1:3) | 20% |
| Fabaceous lecithin | 3% |
| Glycerol | 2.5% |
| Enuatrol | 0.1% |
| PLURONICS F87 | 0.8% |
| Disodium edetate | 0.05% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, PLURONICS F87, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection fabaceous lecithin in [preparation 2] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 7] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 2mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.2% |
| Oil phase (soybean oil: MCT=1:1) | 30% |
| Fabaceous lecithin | 4% |
| Sorbitol | 2.5% |
| Sodium deoxycholate | 0.06% |
| PLURONICS F87 | 1.0% |
| Calcium disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) injection sorbitol, PLURONICS F87, sodium deoxycholate, calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection fabaceous lecithin in [preparation 6] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.0, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 8] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 4mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.4% |
| Oil phase (soybean oil: MCT=1:1) | 30 |
| Lecithin | |
| 5% | |
| Sorbitol | 2.5% |
| Enuatrol | 0.1% |
| PLURONICS F87 | 0.4% |
| Calcium disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) injection sorbitol, PLURONICS F87, sodium deoxycholate, calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 6] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.0, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 9] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 4mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.4% |
| Oil phase (soybean oil: MCT=1:1) | 20% |
| Lecithin | 3% |
| Glycerol | 2.5% |
| Enuatrol | 0.06% |
| Tween 80 | 0.4% |
| Disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 6] are joined in the mixing oil phase of injection MCT, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 10] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05 % |
| Oil phase (Semen Maydis oil: MCT=3:1) | 10% |
| Soybean phospholipid | 1.2% |
| Glycerol | 2.5% |
| Sodium deoxycholate | 0.03% |
| Tween 80 | 0.2% |
| Calcium disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, sodium deoxycholate, calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection soybean phospholipid in [preparation 1] are joined in the mixing oil phase of injection MCT, injection Semen Maydis oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 1,8000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
[prescription 11] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05% |
| Oil phase (soybean oil: safflower oil=1:1) | 10% |
| Lecithin | 1.5% |
| Glycerol | 2.5% |
| Enuatrol | 0.03% |
| Tween 80 | 0.6% |
| Disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 1] are joined in the mixing oil phase of injection safflower oil, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 12] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05% |
| Oil phase (soybean oil) | 10% |
| Lecithin | 1.2% |
| Glycerol | 2.5% |
| Enuatrol | 0.03% |
| Tween 80 | 0.2% |
| PLURONICS F87 | 0.4% |
| Disodium edetate | 0.02% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, PLURONICS F87, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 1] are joined in the injection soybean oil, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 13] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05% |
| Oil phase (soybean oil) | 20% |
| Lecithin | 2.4% |
| Glycerol | 2.5% |
| Enuatrol | 0.08% |
| Tween 80 | 0.3% |
| PLURONICS F87 | 0.3% |
| Disodium edetate | 0.05% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, PLURONICS F87, enuatrol, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 1] are joined in the injection soybean oil, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
[prescription 14] demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid (specification: 0.5mg/ml)
| The demethylate disodium cantharidinate complex | (in cantharidin) 0.05% |
| Oil phase (Semen Maydis oil: safflower oil=1:1) | 20% |
| Soybean phospholipid | 2.4% |
| Glycerol | 2.5% |
| Sodium deoxycholate | 0.08% |
| Tween 80 | 0.3% |
| PLURONICS F87 | 0.3% |
| Disodium edetate | 0.05% |
| Water for injection | Add to 100% |
Preparation method:
(1) glycerol for injection, Tween 80, PLURONICS F87, sodium deoxycholate, disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(2) phosphatide complexes, injection lecithin in [preparation 1] are joined in the mixing oil phase of injection Semen Maydis oil, injection safflower oil, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(3) oil phase is added to aqueous phase, places 2,0000 rmp of high-speed tissue mashing machine to stir 3-5 minute;
(4) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(5) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15min, the rapid cooling of ice-water bath and get final product.
The relevant physicochemical property of the prepared demethylate disodium cantharidinate phosphatide composite liposome of prescription 1-14 microsphere injection liquid is as follows:
| ? | Outward appearance | Particle diameter nm | Content | Envelop rate | Related |
| Prescription | |||||
| 1 | Homogeneous is good | 178.3±45.2 | 100.3% | 85.3% | 0.27 |
| Prescription | |||||
| 2 | Homogeneous is good | 187.6±52.7 | 99.5% | 82.1% | 0.34% |
| Prescription 3 | Homogeneous is good | 173.7±43.2 | 99.1% | 83.4% | 0.25% |
| Prescription 4 | Homogeneous is good | 168.3±46.2 | 99.1% | 81.1% | 0.29 |
| Prescription | |||||
| 5 | Homogeneous is good | 162.8±43.8 | 99.2% | 86.9% | 0.37% |
| Prescription 6 | Homogeneous is good | 165.5±47.1 | 100.7% | 87.6% | 0.29% |
| Prescription 7 | Homogeneous is good | 183.1±59.8 | 98.9% | 81.9% | 0.31% |
| Prescription 8 | Homogeneous is good | 159.5±44.1 | 100.8% | 88.1% | 0.28% |
| Prescription 9 | Homogeneous is good | 168.3±35.8 | 99.4% | 83.4% | 0.29 |
| Prescription | |||||
| 10 | Homogeneous is good | 175.8±48.7 | 97.7% | 82.1% | 0.35% |
| Prescription 11 | Homogeneous is good | 157.2±57.6 | 98.6% | 86.4% | 0.33% |
| Prescription 12 | Homogeneous is good | 189.3±55.1 | 98.7% | 85.1% | 0.27% |
| Prescription 13 | Homogeneous is good | 177.5±42.8 | 100.6% | 87.0% | 0.22% |
| Prescription 14 | Homogeneous is good | 184.2±47.6 | 98.7% | 85.5% | 0.26% |
Embodiment 4 Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid irritation tests
(1) vascular stimulation test
Select 6 of New Zealand white rabbit, 3 white rabbits are in auris dextra auricular vein injection demethylate disodium cantharidinate injection Qs, and left ear is injected 5% aseptic glucose injection of same dose in contrast; 3 white rabbits are in auris dextra auricular vein injection Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid Qz in addition, and left ear is injected 5% aseptic glucose injection of same dose in contrast.Once a day, continuous three days, the last administration was after 24 hours, put to death white rabbit, the response situation of perusal injection site, and dissect rabbit ear blood vessel and surrounding tissue is done paraffin section, dyeing, light microscopy checking.
The result shows: the two kinds of dosage form vascular stimulation tests perusal of demethylate disodium cantharidinate injection is the result show: the blood vessel irritation of Qz is weaker than Qs; Microscopy report shows: the Qs group has to a certain degree blood vessel irritation to New Zealand white rabbit ear blood vessel, and the Qz group has no obvious irritation to New Zealand white rabbit ear blood vessel.
(2) muscle irritation experiment
Select 4 of New Zealand white rabbit, 2 of every kind of dosage forms, respectively at right lateral thigh musculus quadriceps injection Qs and Qz injection 1ml, aseptic 5% glucose injection of left side quadriceps femoris injection equivalent is injected after 48 hours in contrast, puts to death white rabbit, dissect and take out quadriceps femoris, vertically cut, observe the response situation of injection site muscular tissue, determine the order of reaction.
0 grade: unchanged.
1 grade: mild hyperaemia, its scope is below 0.5cm * 1.0cm.
2 grades: moderate is congested, and its scope is more than 0.5cm * 1.0cm.
3 grades: severe is congested, with myodegeneration.
4 grades: necrosis occurs, the brown degeneration is arranged.
5 grades: the popularity necrosis occurs.
Then calculate 4 quadriceps femoris order of reaction summations, again test if the difference of the peak of the quadriceps femoris order of reaction and minimum, then should be got 2 healthy rabbits in addition greater than 2.After obtaining the result, if 4 quadriceps femoris order of reaction summations think then that less than 10 the local irritation test of test sample is up to specification.
The result shows: the muscle irritation of Qz is weaker than Qs.
The experiment of embodiment 5 Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid hemolytics
Get blood 20 ml from the common carotid artery of New Zealand white rabbit, place in the flask, stir gently with Glass rod, after several minutes, except defibrinating, take out blood, add equivalent 5% glucose injection, centrifugal, remove supernatant; The erythrocyte of precipitation adds 5% glucose injection again and cleans, and is centrifugal.So repeatedly until supernatant is transparent, be made into 2% suspension with 5% glucose injection by erythrocytic capacity.
Get 7 of clean tube, number respectively, add successively each liquid in the following table, the 6th pipe does not add test liquid as the blank pipe, the 7th effective distilled water replaces 5% glucose injection, whether shake up, place 37 ℃ of water-baths, observing respectively at 0.5 hour, 1 hour, 2 hours, 3 hours has haemolysis to occur.
The result shows: Qs, Qz have no haemolysis and occur, and the hemolytic experiment of two kinds of dosage forms is all qualified.
The experiment of embodiment 6 Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid anaphylaxis
Get 8 of healthy guinea pigs, two kinds of dosage forms of Qs, Qz each 4, every corresponding test liquid 0.5ml of guinea pig intraperitoneal injection, the next day once, totally three times.The the 14th, the 21 day corresponding reagent liquid 1ml (the dosage conversion is the same) that supplies of lateral vein injection outside the hind paw of every Cavia porcellus after first injection attacks.Each intravenously administrable was observed 2 hours, as two or more person who occurs grabbing in nose, perpendicular hair, cough, the dyspnea is judged to the positive; One if any spasm, gatism, collapse, shock, the phenomena of mortality is judged to the positive.All are normally negative.
The result shows: two kinds of dosage forms of Qs, Qz all meet the anaphylaxis experimental standard.
The research of embodiment 7 Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid rat plasma pharmacokinetics
Dosage regimen
12 rats are divided into two groups at random, and 6 every group, one night of fasting before the experiment.The first group is matched group, in right back vena femoralis injection demethylate disodium cantharidinate injection (2 mgmL
-1), the second group is tested group, in right back vena femoralis injection Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid, respectively at 10 min, 20 min, 30 min, 45 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h eye sockets are got 0.5 mL blood, put in the centrifuge tube of the sharp end of heparinization in advance, centrifugal, the accurate upper plasma 200 μ L that draw add 20 μ L waters for injection, after processing according to a certain method, get 20 μ L sample introductions, calculate the concentration of demethylate disodium cantharidinate in each time point sample with the standard curve on the same day.
The compartment model pharmacokinetic parameter
Average blood drug level data with 3P97 pharmacokinetics routine processes Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid and demethylate disodium cantharidinate injection.Judge that according to AIC and degree of fitting both models belong to, the result shows that the blood drug level data of Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid and demethylate disodium cantharidinate injection injection all meet two compartment models.
Non-compartment model pharmacokinetic parameter
By blood drug level-time measured data Description internal procedure, calculate pharmacokinetic parameter.The result shows, main pharmacokinetic parameters AUC
0-∞, AUMC
0-∞, AUC
0-t, AUMC
0-t, S
2, MRT, the VRT there was no significant difference is seen Fig. 1.
The distribution research of embodiment 8 Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid rat tissues
Dosage regimen
60 rats are divided into two groups at random, 30 every group.The first group is matched group, and in right back vena femoralis injection demethylate disodium cantharidinate injection, the second group is tested group, in right back vena femoralis injection Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid, respectively at 0.5 h, 2 h, 6 h, 12 h, every 6 rats of 24 h(, first, each 3 of second groups) sacrificed by decapitation animal, take out the heart, liver, spleen, lung, kidney, brain, clean with normal saline flushing, blot with filter paper, take by weighing 0.5 g, add 1 mL normal saline homogenate, less than the direct homogenate of the tissue of 0.5 g, centrifugal, get supernatant, after " tissue sample is processed and measured " lower processing, get 20 μ L sample introductions, calculate the concentration of NCTD in each time point sample with the standard curve on the same day.
Experimental result
The meansigma methods of each each Tissue of time point of laboratory animal and the relation of time are seen Fig. 2～7 behind laboratory animal intravenous injection Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid and the Injectio natarii norcantharidatis injection.
Show that by blood plasma pharmacokinetics result of the test in the rat body Injectio natarii norcantharidatis phosphatide composite liposome microsphere injection liquid there is no showing sustained release effect in the rat body.Tissue distribution kinetics is investigated experimental result and is shown that the distribution of demethylate disodium cantharidinate in kidney, liver, heart and blood plasma is more, and in lung, spleen and brain distribution-free almost.After changing dosage form, the total distributed in liver slightly changes, substantially do not change the distribution characteristics of other tissue in the medicine body.
Take the demethylate disodium cantharidinate injection as reference preparation, measure the tissue distribution dynamic characteristic in the rat body, the result shows: after demethylate disodium cantharidinate is prepared into the administration of lipid microsphere posterior vein, do not change medicine rat spleen, lung, kidney, brain, and blood plasma in holdup time and drug distribution amount, the distribution in hepatic tissue slightly increases.
Claims (1)
1. demethylate disodium cantharidinate phosphatide composite liposome microsphere injection liquid, its composition is by weight percentage:
Demethylate disodium cantharidinate phosphatide complexes 0.05% in cantharidin;
Fat-soluble medium 10%;
Glycerol 2.5%;
Sodium deoxycholate 0.03%;
PLURONICS F87 0.2%;
Calcium disodium edetate 0.02%;
All the other are water for injection;
Wherein said fat-soluble medium is the mixture of soybean oil: MCT Oil=3:1;
Its preparation method comprises the steps:
(1) preparation of demethylate disodium cantharidinate phosphatide complexes: remove the methyl disodium cantharidinate, in norcantharidin, 2g, soybean phospholipid 6g adds ethyl acetate 100ml, 60 ℃ after compound 1 hour, rotary evaporation is gone out ethyl acetate, more than 12 hours, namely gets medicine carrying soybean phospholipid complex through vacuum drying under 20-30 ℃, airtight package is put into refrigerator cold-storage and is preserved;
(2) glycerol for injection, PLURONICS F87, sodium deoxycholate, calcium disodium edetate are scattered in an amount of water for injection, are heated to 70 ℃ to the magnetic stirring apparatus to be stirred to whole dissolvings as water;
(3) phosphatide complexes in (1) is joined in the mixing oil phase of injection MCT Oil, injection soybean oil composition, be heated to be stirred under 80 ℃ and dissolve fully as oil phase;
(4) oil phase is added to aqueous phase, places high-speed tissue mashing machine, turn with per minute 1,8000 and stirred 3-5 minute;
(5) with 0.1 molL
-1Sodium hydroxide or hydrochloric acid solution are regulated pH value to 7.5, and the water for injection dilution is settled to recipe quantity, are transferred in the high pressure homogenizer, under 40 ℃, with 800 bar pressure homogenizing 6～10 times;
(6) bottling, sealed cans nitrogen, 121 ℃ of sterilization 15 min, the rapid cooling of ice-water bath and get final product.
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| CN103720653B (en) * | 2012-10-12 | 2016-01-20 | 天津药物研究院 | A kind of vinorelbine submicron emulsion injection and preparation method thereof |
| CN102973503B (en) * | 2012-12-25 | 2015-04-01 | 辽宁正鑫药物研究有限公司 | Norcantharidin derivative lipid microsphere injection and preparation method thereof |
| CN103655471B (en) * | 2013-12-13 | 2015-07-29 | 山东世博金都药业有限公司 | Liver targeting norcantharidin esterified derivatives sub-microemulsion injection and method for making |
| CN112190547B (en) * | 2020-09-30 | 2023-01-03 | 北京诺康达医药科技股份有限公司 | Lipid microsphere composition and preparation method thereof |
| CN112773728A (en) * | 2020-12-31 | 2021-05-11 | 江西科技师范大学 | Compound of nano-encapsulated resveratrol, preparation method and application thereof |
| CN115671048A (en) * | 2022-06-06 | 2023-02-03 | 沈阳信康药物研究有限公司 | Norcantharidin liposome-emulsion complex injection and preparation method thereof |
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| CN1788723A (en) * | 2005-11-20 | 2006-06-21 | 沈阳药科大学 | Liposome microsphere injection liquid containing demethylate disodium cantharidinate and its preparation method |
| CN102068702A (en) * | 2010-12-31 | 2011-05-25 | 苏州大学 | Lactosyl-norcantharidin phospholipid compound and preparation method thereof |
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