CN102973503B - Norcantharidin derivative lipid microsphere injection and preparation method thereof - Google Patents
Norcantharidin derivative lipid microsphere injection and preparation method thereof Download PDFInfo
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Abstract
The invention relates to norcantharidin derivative lipid microsphere injection and a preparation method thereof. The injection contains norcantharidin imide derivative, oil phase, water and surfactant; and the formula of the injection contains the following components by mass: 5 to 30 percent of oil phase, 0.001 to 1 percent of norcantharidin imide N-alkyl derivative, 0.5 to 7 percent of surfactant, 1 to 5 percent of osmotic pressure regulator, and the balance of injection water. Norcantharidin is connected with a long-chain saturated alkane group through amino, so that the lipid solubility of the injection is greatly improved, and the lipid solubility of the injection is improved together with increase of the chain length of the saturated alkane group. Lipid microspheres can carry medicaments with the granularity of more than 90 percent into lipid cores and (or) an interfacial film, so that the medicament loading capacity and the encapsulation efficiency are greatly improved, the vascular stimulation during injection is reduced, the physical and chemical stability of the medicaments and the lipid microsphere injection is improved, the in-vivo acting time of the medicaments is prolonged, the treatment effect is improved, and the toxic or side effect is reduced.
Description
Technical field: the present invention relates to lipid microsphere of a kind of norcantharidin imide N-alkyl derivative and preparation method thereof, belongs to medical art.
Background technology:
Malignant tumor as one of larger public health problem in the whole world, the greatly health of harm humans, and will first killer of the new century mankind be become.In world wide, see global new cancer cases 1,010 ten thousand, dead 6,200,000 in 2000, existing cases of cancer 2,240 ten thousand, 2008, pathogenesis of cancer number and death toll rose to 7,560,000 respectively, and estimating 2015 will have 15,000,000 new cases.Malignant tumor is no longer the serious disease of advanced industrial country simultaneously, and developing country is faced with larger Disease Spectrum.China is as a developing power, and due to industrialization, the existence of urbanization and the problem of an aging population, the situation that malignant tumor faces is also more severe.Therefore, the research and development of antitumor drug become the emphasis of pharmacy worker research.But, because the untoward reaction of most of antitumor drug is serious, in therapeutic process, the body and mind of patient is subject to great injury, therefore, while consideration improves medicine effect, reduce the zest of medicine, toxicity and untoward reaction, the compliance of raising patient medication and life quality become the most important thing of research.
Norcantharidin (norcantharidin, NCTD) being the synthesis of derivatives of cantharidin, is the new drug of the Hepatoma therapy first developed by China, compared with cantharidin, not only significantly improve anticancer effect, and greatly reduce nephrotoxicity and the intense stimulus to urinary system.It has the advantage of its uniqueness in treatment tumor, comprise leukocyte increasing, immunity moderation and do not produce bone marrow depression, it is the first-selected ancillary drug of the multiple alimentary tract cancers such as Hepatoma therapy, the esophageal carcinoma, gastric cancer and carcinoma of gastric cardia, wherein based on primary hepatocarcinoma, all can use before and after operation and chemicotherapy.Clinical application shows, and to liver cancer patient, the use of norcantharidin can make life span extension, and AFP declines, and the liver retraction of tumor body and enlargement, quality deliquescing, can alleviate hepatalgia and ascites; For the esophageal carcinoma, gastric cancer, patients with cardiac cancer, visible cancer is stable or reduce, pain and block alleviation, and feed improves.Norcantharidin can also improve liver function, is also the medicine for treatment of hepatitis, liver cirrhosis and hepatitis B virus carriers simultaneously.These advantages make norcantharidin have good application prospect.The norcantharidin used clinically is mainly based on tablet and injection.
But norcantharidin has certain restriction in clinical practice.First, although norcantharidin greatly reduces the toxicity of cantharidin, but do not eliminate completely, still there is urinary system toxicity to a certain degree, zoopery shows, and when dosage arrives to a certain degree, pathological change appears in kidney and liver, during Clinical practice there is untoward reaction more than 20mg or oral every day more than during 30mg at intravenously administrable in some patients every day, and oral untoward reaction main manifestations is the symptom such as Nausea and vomiting, dizziness.Thus, clinical maximum application dosage also exists strict restriction, this also seriously hinders the performance of curative effect of medication.Second, with 3H-Demethylcantharidin Tablets to mouse stomach research display, norcantharidin is comparatively fast distributed in each tissue after absorbing, within after administration 15 minutes, at liver, cancerous tissue reaches peak concentration, and after 6 hours, concentration significantly declines, in 24 hours, major part is through renal excretion, less savings in body, release rate is very fast in vivo for this result of study display norcantharidin, and the compliance of patient medication is reduced.In addition, distribution in vivo result shows, after mouse stomach Injectio natarii norcantharidatis solution, and AUC
0-tvalue is descending to be followed successively by: small intestinal, stomach, uterus, kidney, testis, liver, the heart, brain, widely distributed in vivo after prompting norcantharidin oral administration, and distributing less and release rate is fast in liver organization, this kind of characteristic not only reduces drug effect, and adds the toxicity to other organs.3rd, the existing norcantharidin injection applied clinically mostly is sodium salt (5mgmL
-1, 2ml) pH about 9.0, much larger than pH value of blood (7.35 ~ 7.45), therefore blood vessel wall is stimulated comparatively large, easily causes chemical phlebitis; Because medicine itself also has stronger zest, also can cause vasospasm, blood reduces, and the concentration of local Injectio natarii norcantharidatis increases relatively, and increase the weight of phlebitis, if transfusion speed is much larger than velocity of blood flow, then phlebitic incidence rate obviously increases.Main adverse reaction shows as and easily causes inflammation of vein, and vein portion is red, swollen, hot, bitterly, rectangular rubescent along vein traveling, if any oozing out, then red and swollen one-tenth is block, and limitation of movement takes a turn for the better through hot packing process and cures.Therefore, clinical for periphery superficial intravenous infusion time, the ratio of superficial phlebitis occurs quite high, some patient's refusals are treated with this medicine, affect medicine and normally use clinically.
In view of above-mentioned characteristic, play better play curative effect to make norcantharidin, the exploitation that can give full play to the derivant of efficient, low toxicity, the low irritant of norcantharidin effect and novel form undoubtedly tool is of great significance.
Intravenous injection fatty emulsion, as the carrier of antitumor drug, is widely used recently.Its special physicochemical property and hypotoxicity determine it can as fat-soluble medicine, the particularly good carrier of cancer therapy drug, anaesthetic and anti-inflammatory drug.Compare with conventional injection liquid phase, lipomul can reduce blood vessel irritation, and plays the effect of target tumor tissue.Intravenous injection fatty emulsion is applied to clinical nearly half a century as the important supply mode of the outer energy of intestinal, is the composition according to Chylomicron, structure and feature and designing, the emulsion grain solution be mainly made up of triglyceride and phospholipid.Because pharmaceutical pack is wrapped in the lipid core part of lipomul, this similar is in microsphere, and therefore the title of lipid microsphere (LipidMicrosphere, LM) is also applied and gives birth to.It is generally acknowledged that lipid microsphere is by being dissolved in fatty oil by medicine, and make after aqueous phase through phospholipid emulsion dispersion, be a kind ofly be soft substrate with fatty oil and the microparticulate system encapsulated by immobilized artificial membrane, as pharmaceutical carrier, there is much physical chemistry and advantage biologically:
1. are good carriers of fat-soluble medicine.Clinical many medicine poorly water-solubles, must rely on the effect of organic solvent competence exertion, and organic solvent itself have certain toxicity, also may interference medicament effect.
2. effectively can increase the stability of medicine.In pastille lipid microsphere, there is quite a few drug distribution in oil phase or oil-water interfacial film, avoid medicine and directly contact with water.For facile hydrolysis or the medicine to pH sensitive, this isolation serves the effect increasing stability.
3. drug moiety is wrapped in oil phase or interfacial film, avoids and contacts with the direct of body fluid, because this reducing the issuable local of medicine self and blood vessel irritation.In addition, medicine is by slow releasing in oil phase in vivo, can avoid the untoward reaction that medicine causes due to initial stage excessive concentration when injecting.
4. particle diameter about 200nm small particle can engulf by the phagocyte of reticuloendothelial system and be trapped in reticuloendothelial system (as liver, lung), have passive targeting, this feature improves drug effect for antitumor drug, reduction toxic and side effects is particularly important.
5., during carrier as antitumor drug, the effect of target tumor tissue can be played.Change relevant to the targeting of tumor tissues with the organizational structure of these diseased regions.This is due at growth of cancer cells position; pathological changes causes the permeability of blood capillary to increase; the easier capillary wall by diseased region of lipid microsphere also infiltrates in tumor tissues; and the capillary permeability of normal structure is very little; and do not allow lipid microsphere granule to pass through, thus reduce the distribution of medicine in normal structure.Meanwhile, due to the poor structural integrity of solid tumor mass, lymphatic return lacks, and makes lipid microsphere be trapped in tumor tissues, can greatly reduce the toxicity of medicine normal tissue.
The related documents report of the people such as Takahashi: win mycin with Lay and manufacture experimently into LM, and with injection in contrast, test with tumor-bearing rat, carry out vein, then tumor size and tumor drug concentration is measured respectively, result shows: the tumor regression more than 50% of LM group, still can detect medicine in after administration 10 hours; And the tumor of injection is without obviously reducing, can not detecting medicine after 30 minutes to 1 hour, more than indicating LM and have selective distribution and discharge feature slowly, especially for liver, lymphoid tissue systemic disease and cancer chemotherapy are all good vehicles.
And norcantharidin is slightly water-soluble, in oil, dissolubility is very low, and the sodium salt that its hydrolysis is formed also is only dissolve in minority organic solvent (as acetone, ethyl acetate) and hot water, and the dissolubility in cold water and oil is extremely low.Which results in and norcantharidin or its sodium salt are prepared into LM have certain difficulty, it is not high that research shows that sodium norcantharidate lipid microsphere exists envelop rate, and drug loading is low, and the problem of easy to leak in sterilizing and storage process.
Summary of the invention:
Goal of the invention: the invention provides a kind of Norcantharidin derivative lipid microsphere injection and preparation method thereof, its object is to solve norcantharidin drug loading low, and the problem of easy to leak in sterilizing and storage process, reduce its toxicity and improve its curative effect.
Technical scheme:
A kind of lipide microsphere injection containing norcantharidin imide N-alkyl derivative, it is characterized in that: this infusion pump contains following composition: norcantharidin imide N-alkyl derivative, fat-soluble medium, water and surfactant, in the quality of the pharmaceutical preparations, formula consists of:
Oil phase 5% ~ 30%,
In the norcantharidin imide N-alkyl derivative 0.001% ~ 5% of norcantharidin,
Surfactant 0.5% ~ 7%,
Osmotic pressure regulator 1% ~ 5%,
All the other are water for injection.
In described injection, norcantharidin imide N-alkyl derivative general structure is the compound of I;
Wherein: R is C for being selected from general formula
nh
2n+1straight chain saturated alkane base; N is an integer in 8-24.
In described injection, osmotic pressure regulator is selected from one or more mixture in glycerol, sorbitol, mannitol, glucose.
Oil phase is selected from one or more mixture in mineral oil, vegetable oil, animal oil, quintessence oil and artificial oil; Surfactant is selected from phospholipid, tween, span, F68, one or more mixture in enuatrol, oleic acid, cholic acid, deoxycholic acid.
Described oil phase is selected from one or more mixture in soybean oil, safflower oil, Semen Maydis oil, MCT Oil, pearl barley oil, Petiolus Trachycarpi oil, Oleum Helianthi, Fructrs Hippophae seed oil, Radix Angelicae Sinensis oil, Fructus Zanthoxyli oil, Oleum Bulbus Allii, Oleum Curcumae or Rhizoma Chuanxiong oil.
Described tween is selected from the one in polysorbas20, polysorbate40, polysorbate60, Tween 80, polysorbate85; Described F68 is F68 F68; Described phospholipid is selected from fabaceous lecithin, lecithin or is both mixture.
Containing metal-chelator, antioxidant, buffer salt, pH adjusting agent in preparation; In the quality of the pharmaceutical preparations, metal-chelator is 0% ~ 1%, and antioxidant consumption is 0 ~ 2%, and buffer salt consumption is 0 ~ 2%, and pH adjusting agent consumption is 0.005% ~ 2%.
Metal-chelator is one in disodium edetate, sodium calcium edetate or is both mixture; Antioxidant is one or more mixture in vitamin E, vitamin C, sodium sulfite, sodium pyrosulfite, cysteine; Buffer salt is one or more mixture in acetic acid, sodium acetate, citric acid, sodium citrate, sodium hydrogen phosphate, sodium dihydrogen phosphate; PH adjusting agent is the one in sodium chloride, sodium hydroxide, hydrochloric acid.
A kind of preparation method of the lipide microsphere injection containing norcantharidin imide N-alkyl derivative as above, it is characterized in that: by the above-mentioned quality of the pharmaceutical preparations, after norcantharidin imide N-alkyl derivative is mixed with fat-soluble medium, be redispersed in aqueous phase, prepare after colostrum through high-speed stirred, carry out high pressure homogenize process again, preparation adopts the sterilizing of high-pressure rotary sterilization.
The method is undertaken by following processing step:
Injection osmotic pressure regulator, surfactant, metal-chelator and water soluble antioxidants are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
By the norcantharidin imide N-alkyl derivative of recipe quantity and being dissolved in the oil phase of containing oil soluble antioxidant and oil phase solubilizing agent, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
Under organizing at a high speed dispersion machine to stir, by aqueous phase and oil phase mixing, obtained colostrum;
By colostrum sodium hydroxide or hydrochloric acid solution adjust ph to about 4.0-9.0, be then settled to recipe quantity with water for injection, be transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 20-80MPa pressure homogenizing 4-15 time with ice-water bath;
Fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly, and after lamp inspection is qualified, packaging, in 4-10 ° of C storage.
Advantageous effect:
Preparation of the present invention utilizes excellent fat-soluble of norcantharidin imide N-alkyl derivative, and the lipid core 90% pharmaceutical pack being wrapped in lipid microsphere is inner, and entrapment efficiency and drug loading are improved greatly.After imide N-alkyl derivative is prepared into lipid microsphere, significantly reduce the zest of blood vessel when showing as injection after administrated by injection, preparation shows as good chemical physical stability in storage process, without the Seepage of medicine.Pharmacokinetics in rats result shows, and imide N-alkyl derivative can be hydrolyzed to norcantharidin by amidase in vivo, medicine is extended action time in vivo, plays slow releasing function, efficient Tumor suppression growth.
Norcantharidin imide N-alkyl derivative lipide microsphere injection (oil in water emulsion) prepared by the present invention, blood vessel irritation when reducing injection, improve the physical and chemical stability of medicine and preparation, action time in prolong drug body, improve curative effect and reduce toxic and side effects, thus there is certain novelty and stronger practicality.
Accompanying drawing illustrates:
Fig. 1 is LM structural representation;
Fig. 2 is rat plasma drug concentration-time graph in embodiment 9.
Embodiment:
Central scope of the present invention is: provide a kind of lipide microsphere injection (compositions of oil-in-water emulsion) containing norcantharidin imide N-alkyl derivative, and provide preparation method.Be intended to reduce medicine to the zest of blood vessel, prolong drug action time in vivo, alleviate its nephrotoxicity, the not high and leakage problems of the entrapment efficiency simultaneously utilizing the fat-soluble solution Injectio natarii norcantharidatis lipid microsphere of derivant to cause due to its highly-water-soluble.
This infusion pump contains: norcantharidin imide N-alkyl derivative, fat-soluble medium, water and surfactant.
Formula of the present invention consists of: (in the quality of the pharmaceutical preparations),
Oil phase 5% ~ 30%;
Norcantharidin imide N-alkyl derivative 0.001% ~ 5%(is in norcantharidin);
Surfactant 0.5% ~ 7%;
Osmotic pressure regulator 1.0% ~ 5%;
All the other are water for injection.
Use term " oil " to represent the acceptable material of a large class physiology herein, be selected from mineral oil, vegetable oil, animal oil, quintessence oil and artificial oil, and composition thereof.Therefore, fat-soluble medium has material very of different nature in order to what refer to a wide region.When with type or functional classification oil, as mineral oil source comprises fat or cerul hydrocarbon, the fat of aromatic hydrocarbon or mixing and aromatic radical hydrocarbon from oil.Also the oil of petroleum derivation is comprised as refined paraffin wax wet goods in mineral oils is than two.In vegetable oil classification, oil is mainly derived from seed or nut, and comprises drying oil as Semen Lini and Oleum Verniciae fordii; Semi-drying oil is as safflower oil and soybean oil; Non-drying oil is if Oleum Ricini, Oleum Gossypii semen and Oleum Cocois and the soap stock that can be used as are as Petiolus Trachycarpi oil and Oleum Cocois.In animal oil classification, oil usually from as sebum, Adeps Sus domestica and stearic fat.Liquid animal oil comprises fish oil, oleic acid, spermaceti wet goods.They comprise abundant fatty acid usually.Comprise some vegetable oil, as olive oil, Oleum Gossypii semen, corn oil and Oleum Arachidis hypogaeae semen, also comprise the fish oil that some are special, they are widely used as medicine owing to being rich in vitamin, as morrhua liver, and shark hepatic wet goods.Aqueous fatty oil is as single, double, triglyceride, or its mixture is preferred oil.According to the present invention, the triglyceride of medium chain are also useful oil.Preferably long-chain fat acid glyceride, medium chain length fatty acid triglyceride, and composition thereof.
Surfactant used can be any surfactant, usually has phospholipid, tween, span, F68, enuatrol, oleic acid, cholic acid, deoxycholic acid, and composition thereof, wherein, described tween is selected from polysorbas20, polysorbate40, polysorbate60, Tween 80, polysorbate85, and composition thereof; Described F68 is F68 F68; Described phospholipid is selected from fabaceous lecithin, lecithin or is both mixture.
The osmotic pressure regulator used in invention is selected from glycerol, sorbitol, mannitol, glucose, and composition thereof.
In described injection, norcantharidin imide N-alkyl derivative general structure is the compound of I;
Wherein: R is C for being selected from general formula
nh
2n+1straight chain saturated alkane base;
N is an integer in 8-24.
If needed, can add multiple additives in compositions, as contained metal-chelator, the general metal-chelator used is disodium edetate, sodium calcium edetate, and composition thereof, consumption counts 0% ~ 1% with preparation consumption; Antioxidant be vitamin E, vitamin C, sodium sulfite, sodium pyrosulfite, cysteine, and composition thereof, consumption counts 0 ~ 2% with preparation consumption; Buffer salt is acetic acid, sodium acetate, citric acid, sodium citrate, sodium hydrogen phosphate, sodium dihydrogen phosphate, and composition thereof, consumption counts 0 ~ 2% with preparation consumption.PH adjusting agent is sodium chloride, sodium hydroxide, hydrochloric acid, commonly used buffer salt etc., and one or more mix pH adjusting agent, and consumption counts 0.005% ~ 2% with preparation consumption.
The present invention also designs the preparation method of the lipide microsphere injection (oil in water emulsion) containing norcantharidin imide N-alkyl derivative, specifically comprises following operating procedure:
A. injection osmotic pressure regulator, surfactant, metal-chelator and water soluble antioxidants are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
B. by the norcantharidin imide N-alkyl derivative of recipe quantity and being dissolved in the oil phase of containing oil soluble antioxidant and oil phase solubilizing agent, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
C. under organizing at a high speed dispersion machine to stir, by aqueous phase and oil phase mixing, obtained colostrum;
D. by colostrum sodium hydroxide or hydrochloric acid solution adjust ph to about 4.0-9.0, be then settled to recipe quantity with water for injection, be transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 20-80MPa pressure homogenizing 4-15 time with ice-water bath;
E. fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
F. lamp inspection qualified after, packaging, in 4-10 ° of C storage.
Preparation of the present invention utilizes excellent fat-soluble of norcantharidin imide N-alkyl derivative, and the lipid core 90% pharmaceutical pack being wrapped in lipid microsphere is inner, and entrapment efficiency and drug loading are improved greatly.After imide N-alkyl derivative is prepared into lipid microsphere, significantly reduce the zest of blood vessel when showing as injection after administrated by injection, preparation shows as good chemical physical stability in storage process, without the Seepage of medicine.Pharmacokinetics in rats result shows, and imide N-alkyl derivative can be hydrolyzed to norcantharidin by amidase in vivo, medicine is extended action time in vivo, plays slow releasing function, efficient Tumor suppression growth.
Norcantharidin imide N-alkyl derivative lipide microsphere injection (oil in water emulsion) prepared by the present invention, blood vessel irritation when reducing injection, improve the physical and chemical stability of medicine and preparation, action time in prolong drug body, improve curative effect and reduce toxic and side effects, thus there is certain novelty and stronger practicality.
Below in conjunction with example, the present invention is conducted further description.Following examples are only several specific embodiment of the present invention, but design concept of the present invention is not limited thereto.
The norcantharidin imide N-alkyl derivative lipide microsphere injection prepared in following embodiment has the structure similar to traditional liposomal microsphere, with micromolecule phospholipid for emulsifying agent, having with fatty oil is soft substrate, by the spherical microstructure that the monofilm of phospholipid molecule is wrapped to form, accompanying drawing 1 is shown in by schematic diagram, and wherein 1 is aqueous phase, and 2 is oil phase, 3 is phospholipid molecule, and 4 is drug molecule.But it compares the drug loading that norcantharidin lipid microsphere has better stability and Geng Gao, change due to structure makes its dissolubility in various oil medium greatly raise, and the aliphatic chain in its structure more easily inserts in phospholipid molecule layer, the drug distribution of more than 90% is in oil phase, not easily be transferred to aqueous phase, significantly improve its drug loading, strengthen its stability.
Method in following examples, if no special instructions, is conventional method.
Percentage composition in following examples, if no special instructions, is mass percentage.
The formula preparation technique of embodiment 1 norcantharidin imide N-alkyl derivative (n=8-12) lipide microsphere injection.
Norcantharidin imide N-alkyl derivative R used in embodiment 1 is C
8-12saturated straight chain alkyl.
[prescription 1] specification: 0.1mg/10ml (in norcantharidin)
Preparation method 1:
(1) glycerol for injection, Tween-80, EDTA, enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin amine derivant of recipe quantity with in being dissolved in containing lecithin injection Oleum Fructus Bruceae, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 2] specification: 500mg/10ml (in norcantharidin)
Preparation method 2:
(1) glycerol for injection, PLURONICS F87, EDTA are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin amine derivant of recipe quantity with in being dissolved in containing fabaceous lecithin and oleic acid injection soybean oil, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, oil phase is added to aqueous phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 3] specification: 30mg/10ml (in norcantharidin)
Preparation method 3:
(1) glycerol for injection, Tween-80, PLURONICS F87, EDTA and enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity with in being dissolved in containing lecithin medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
The formula preparation technique of embodiment 2 norcantharidin imide N-alkyl derivative (n=13-18) lipide microsphere injection
Norcantharidin imide N-alkyl derivative R used in embodiment 2 is for being selected from C
13-18saturated straight chain alkyl.
[prescription 4] specification: 0.1mg/10ml (in norcantharidin)
Preparation method 1:
(1) glycerol for injection, PLURONICS F87, EDTA are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity with in being dissolved in containing lecithin and oleic acid injection safflower oil, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 5] specification: 500mg/10ml (in norcantharidin)
Preparation method 2:
(1) glycerol for injection, PLURONICS F87, Tween-80, EDTA and enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity and being dissolved in containing in the injection soybean oil of fabaceous lecithin and the mixing oil phase of medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, oil phase is added to aqueous phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 6] specification: 30mg/10ml (in norcantharidin)
Preparation method 3:
(1) glycerol for injection, PLURONICS F87, EDTA and enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity with in being dissolved in containing lecithin mixing oil phase that penetrating with soybean oil and medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
The formula preparation technique of embodiment 3 norcantharidin imide N-alkyl derivative (n=19-24) lipide microsphere injection;
Norcantharidin imide N-alkyl derivative R used in embodiment 3 is for being selected from C
19-24saturated straight chain alkyl.
[prescription 7] specification: 0.1mg/10ml (in norcantharidin)
Preparation method 1:
(1) glycerol for injection, PLURONICS F87, EDTA are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) the norcantharidin imide N-alkyl derivative of recipe quantity and oleic acid are dissolved in containing in lecithin and the injection safflower oil of oleic acid and the mixing oil phase of medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 8] specification: 500mg/10ml (in norcantharidin)
Preparation method 2:
(1) glycerol for injection, Tween-80, sorbitan fatty acid ester-80, EDTA and enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity and being dissolved in containing in the injection soybean oil of fabaceous lecithin and the mixing oil phase of medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, oil phase is added to aqueous phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
[prescription 9] specification: 30mg/10ml (in norcantharidin)
Preparation method 3:
(1) glycerol for injection, PLURONICS F87, EDTA and enuatrol are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80 ° of C, and be stirred to whole dissolving, as aqueous phase;
(2) by the norcantharidin imide N-alkyl derivative of recipe quantity with in being dissolved in containing lecithin mixing oil phase that penetrating with soybean oil and medium chain triglyceride, put in magnetic stirring apparatus and be heated to 30-80 ° of C to mix homogeneously, as oil phase;
(3), under organizing at a high speed dispersion machine to stir, aqueous phase is added to oil phase, first slow rear fast, stir 2 times, each 3min, obtained colostrum;
(4) by colostrum 0.1molL
-1sodium hydroxide or hydrochloric acid solution adjust ph, to about 4.0-9.0, are then settled to recipe quantity with water for injection, are transferred in high pressure homogenizer, control homogenizing temperature at 30-80 ° of C, to be about 70MPa pressure homogenizing 4-15 time with ice-water bath;
(5) fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly.
(6) lamp inspection qualified after, packaging, in 4-10 ° of C storage.
Except the compositions listed by above-mentioned prescription, prepare several formulations according to above method, they composed as follows:
Embodiment 4 norcantharidin imide N-alkyl derivative lipide microsphere injection stability test
Because this product is thermodynamic unstable system, belong to heat sensitive dosage form.Explanation according to Chinese Pharmacopoeia version crude drug in 2010 and pharmaceutical preparation stability test guideline: heat sensitive medicine accelerated test is placed 6 months under the condition of 25 ± 2 DEG C, place 12 months under the condition of 6 ± 2 DEG C, investigate the change of physical and chemical stability.This experiment is only to above-mentioned prescription 3,6, and 9 investigate.
25 ± 2 DEG C, table 1 prescription 3 sample accelerates 6 months result of the tests
6 ± 2 DEG C, table 2 prescription 3 sample keeps sample 12 months result of the tests for a long time
25 ± 2 DEG C, table 3 prescription 6 sample accelerates 6 months result of the tests
6 ± 2 DEG C, table 4 prescription 6 sample keeps sample 12 months result of the tests for a long time
25 ± 2 DEG C, table 5 prescription 9 sample accelerates 6 months result of the tests
6 ± 2 DEG C, table 6 prescription 9 sample keeps sample 12 months result of the tests for a long time
Acceleration and long-term shelf-stability result of the test show: norcantharidin imide N-alkyl derivative lipide microsphere injection is under 50ml low Pyrex silication Coated bottle injects the terms of packing of liquid chlorinated butyl rubber bung added with antibiotic bottle aluminium-plastic combined cover, assay through 25 ± 2 DEG C of accelerated tests 6 months and 6 ± 2 DEG C of long term tests, 12 months every inspection target still meets quality standard regulation, illustrates that this terms of packing can ensure the steady quality of preparation at duration of storage.
Embodiment 5 norcantharidin imide N-alkyl derivative lipide microsphere injection is investigated inhibiting tumour cells effect.
The external inhibitory action to Human leukemia cell line HL-60 of norcantharidin imide N-alkyl derivative lipide microsphere injection is measured by mtt assay.Take the logarithm the cell of trophophase, adjust suitable cell density, be inoculated in 96 orifice plates, 100 μ l/well, are incubated at 37 DEG C, in the incubator of 5%CO2.Add certain density pastille lipide microsphere injection after cultivating 24h to hatch altogether, after drug effect 48h, cell and 0.25mg/ml MTT are jointly hatched 3-4h at 37 DEG C, centrifugal absorption supernatant, add 100 μ l dimethyl sulfoxide (DMSO), after dissolving completely, use microplate reader to measure its OD value in 492nm.Last with blank group OD value for 100%, calculate each group of cell inhibitory rate.
Set up blank group, administration group and positive controls separately, wherein, blank group is blank liposomes microsphere injection liquid, and administration group is the pastille lipide microsphere injection of different chain length, and positive controls is etoposide inj, and often group establishes 4 multiple holes.
Table 7 Norcantharidin derivative lipid microsphere injection is to survival suppression ratio (%) (Mean ± SE) of HL-60 cell strain
The above results shows, norcantharidin imide N-alkyl derivative lipide microsphere injection has good lethal effect to tumor cell.
Embodiment 6 norcantharidin imide N-alkyl derivative lipide microsphere injection irritation test
" chemicals stimulation, anaphylaxis and hemolytic investigative technique guideline [H] GPT4-1 " the setting experiment of promulgating according to national Yao Shen center adopts one-level rabbit, according to clinical adult's intravenous injection Norcantharidin sodium injection chemotherapy regimen, norcantharidin imide N-alkyl derivative lipide microsphere injection group (by reagent group), Norcantharidin sodium injection (positive drug group), lipide microsphere injection group (group of solvents) and normal saline group (negative control group) rabbit auricular vein per injection dosage are 0.6mg/kg.
Get healthy one-level rabbit 10, male, average weight 2.42 ± 0.11kg.4 groups of rabbit ears are established in experiment, namely by reagent group, positive drug group, negative control group and group of solvents, often organize 5 rabbit ears.Once, for three days on end, last administration is after 24 hours in injection every day, puts to death white rabbit, the response situation of perusal injection site, and dissects rabbit ear blood vessel and surrounding tissue does paraffin section, dyeing, light microscopy checking.The result of perusal injection site reaction situation is as shown in table 8, and pathological section is checked by disease prevention and control center of Liaoning Province, and presents audit report.Observe after the injection of rabbit auricular vein and record injection site irritation reactions change, and carry out scoring and stimulus intensity evaluation according to each experimental group irritant reaction intensity.
Local excitation reaction standards of grading: be 0(-without scores of erythema); Slight scores of erythema is 1(+); Moderate erythema scoring is 2(++); Severe scores of erythema is 3(+++); Purple erythema forms scoring for 4(++++ to slight eschar).Mark as 0(-without edema); Mild edema scoring is 1(+); Intermediate edema scoring is 2(++); Severe edema's scoring is 3(+++); Severe edema scoring is 4(++++).
Injection site irritation intensity evaluation standard: the total score value of each experimental group local nonirritant is 0-0.49; The total score value of slight zest is 0.5-2.99; The total score value of moderate zest is 3.0-5.99; The total score value of strong and stimulating is 6.0-8.0.
Table 8 rabbit ear edge intravenous site Acute response stage local excitation reaction scoring and stimulus intensity compare (n=5 the rabbit ear)
Table 8 shows, and after rabbit auricular vein injection positive drug, early stage injection site is along the visible subcutaneous erythema of blood vessel traveling, and have no obvious edema and occur, this group local excitation intensity is 1.5.Along with time lengthening after administration, the reaction of this group injection site irritation alleviates gradually.Visible slightly rubescent by injection site early stage after the administration of reagent group, inject and within latter 4 days, return to gradually normally.Group of solvents and normal saline group rabbit ear injection site skin have no obvious stimulation reaction.
Rabbit auricular vein injection site pathological examination result display: group of solvents and normal saline group rabbit auricular vein tissue have no obvious pathological changes.1d after positive drug group intravenous injection Injectio natarii norcantharidatis injection, 3 rabbit ear auricular vein expansions, congestion, 1 rabbit ear vein peripheral tissue edema, has no inflammatory cell infiltration.By 1d after reagent group intravenous injection norcantharidin imide N-alkyl derivative lipide microsphere injection, 2 rabbit ear auricular vein expansions, congestion, 1 visible mild inflammatory cellular infiltration of rabbit ear vein surrounding tissue.
By the inspection to test sample, comprehensive analysis injection site irritation experimental result prompting, after intravenously administrable, Norcantharidin sodium injection has blood vessel irritation to a certain degree to injection site blood vessel and tissue, and norcantharidin imide N-alkyl derivative lipide microsphere injection has no obvious stimulation to injection site local vascular and tissue.
Embodiment 7 norcantharidin imide N-alkyl derivative lipide microsphere injection hemolytic is tested
Promulgate " chemicals stimulation, anaphylaxis and hemolytic investigative technique guideline [H] GPT4-1 " according to national Yao Shen center.The experiment of norcantharidin imide N-alkyl derivative lipide microsphere injection hemolytic adopts hemolytic experiment and hemolysis in vitro experiment in body to carry out observing (with reference to the long malicious experimental result of this test medicine) respectively.In body, hemolytic experiment adopts SD rat.Hemolysis in vitro experiment employing 2% rabbit erythrocyte suspension and gradient concentration test medicine mixed liquor are measured by reagent various dose pipe supernatant optical density value after placing 37 DEG C of water-bath different times.
In body, hemolytic experiment adopts SD rat, experiment setting blank liposomes microsphere injection liquid group (abbreviation negative control group), Injectio natarii norcantharidatis injection (being called for short positive drug group) and norcantharidin imide N-alkyl derivative lipide microsphere injection group (being called for short by reagent group), often organize 8 rats.Adopt the administration of tail vein injection method, administration group rat unit body weight dose is 0.6mg/kg, at interval of 1d tail vein injection once, injects 6 times altogether.Negative control group tail vein injection equal-volume blank liposomes microsphere injection liquid.After last administration, 2d respectively organizes rat through femoral artery sacrificed by exsanguination, gets blood, urinates and carry out red blood cell count(RBC), packed cell volume, reticulocyte count, urine bilirubin and urobilinogen mensuration respectively.In body, hemolytic test observed result is in table 9.
Hemolytic experiment blood and uroscopy results contrast (n=8) in each experimental group body of table 9
Note: (1) positive drug group and comparing with negative control group respectively by reagent group, a:p>0.05;
(2) compare with positive drug group by reagent group, b:p>0.05.
Table 9 display shows by hemolytic experiment result in reagent and positive drug rat body: positive drug group rat blood red blood cell count(RBC) after intravenously administrable, packed cell volume, reticulocyte count mensuration average compare all without significant difference with negative control group respectively, and to compare with negative control group by the above-mentioned observation index of reagent group and be showed no significant difference.
Hemolysis in vitro experiment employing 2% rabbit erythrocyte suspension and gradient concentration test medicine mixed liquor are measured by reagent various dose pipe supernatant optical density value after placing 37 DEG C of water-bath different times.Positive pencil, tested pencil, solvent pipe and negative control pipe are established in experiment.Tested pencil and positive pencil adopt 5 pipe red cell suspensions to observe respectively, and each pipe test medicine reaction density is respectively 0.06mg/ml, 0.12mg/ml, 0.18mg/ml, 0.24mg/ml, 0.30mg/ml.Solvent pipe and negative control pipe add equal-volume blank liposomes microsphere injection liquid and normal saline respectively.Hemolysis in vitro EXPERIMENTAL DESIGN and haemolysis criterion are in Table 10-11 laboratory observation and compare this by reagent and positive drug to the impact of isolated rabbit erythrocyte haemolysis.Test medicine hemolysis in vitro experimental observation the results are shown in Table 12-13.
Table 10 hemolytic test design table
Note: norcantharidin imide derivative lipide microsphere injection application concentration is 3mg/ml.
Table 11 hemolytic test result criterion
A situation arises (n=4) for table 12 perusal different pharmaceutical dosimetry pipe supernatant haemolysis
The different minute of table 13 and different pharmaceutical dosimetry pipe supernatant measure optical density value (OD value) and compare (n=4)
Note: compare with negative control group respectively by reagent group and positive drug group detector tube OD value and have no significant difference.
Result shows: various dose test medicine and positive drug rabbit red cell suspension after 37 DEG C of water-bath differential responses time does not all find obvious haemolysis.
By the inspection to test sample, result is pointed out, and norcantharidin imide N-alkyl derivative injection and Injectio natarii norcantharidatis injection are showed no obvious haemolysis in vivo with under hemolysis in vitro experiment condition.
Embodiment 9 norcantharidin acyl imide N-alkyl derivative lipide microsphere injection rat plasma pharmacokinetics is tested
Norcantharidin imide N-alkyl derivative lipide microsphere injection pharmacokinetic studies adopts with Injectio natarii norcantharidatis injection as contrast, investigate the pharmacokinetics of norcantharidin imide N-alkyl derivative injection in rat body, calculated main pharmacokinetic parameters, for the clinical research of this preparation provides reference.And respectively the calculating of compartment model and non-compartment model pharmacokinetic parameter is carried out to two kinds of animal vivo test results and analytic demonstration is carried out to experimental phenomena and result, investigate the metabolic condition of medicine in rat body under different dosing dosage conditions with this.
Clinical pharmaceutical quantities of recommending is for once-a-day 10 ~ 30mg.Determine that in rat, dosage group is that 2.7mg/kg(is in norcantharidin with reference under " chemicals Non-clinical Pharmacokinetics investigative technique guideline " dosage choice item).
12 rats are divided into 2 groups at random, often organize 6, fasting one night before experiment, first group is matched group, in rear vena femoralis injection Injectio natarii norcantharidatis injection (5mg/ml), second group is tested group, in right back vena femoralis injection norcantharidin imide N-alkyl derivative lipide microsphere injection (5mg.ml (in norcantharidin)), blood 0.5ml is got respectively at 5min, 15min, 30min, 45min, 1h, 2h, 4h, 6h, 8h, 12h, 24h eye socket, put in heparinization centrifuge tube centrifugal, get upper plasma.After pre-treatment, measure drug level by LC-MS/MS.Measurement result is shown in Fig. 2, and the plasma drug concentration data of application DAS2.0 statistical computation software (mathematics pharmacology Professional Committee of Chinese Pharmacological Society) pharmacokinetics program to norcantharidin imide N-alkyl derivative lipide microsphere injection, Injectio natarii norcantharidatis injection processes.Pharmacokinetic parameters is in table 14.
Table 14. statistical moment estimation demethylcantharidin imide N-alkyl derivative lipide microsphere injection (A1) and the nor-speckle of injection
Every pharmacokinetic parameters of Cantharis acid sodium (R1)
Adopt SPSS software to analyze main pharmacokinetic parameters, each pharmacokinetic parameters of test group and matched group all has significant difference.Contrasted from upper table: the blood drug level of (1) this preparation is moderate, can play curative effect rapidly, can avoid that R1 blood drug level is too high brings safety risks.(2) this preparation internal metabolism speed is moderate, compared with R2, has slow release effect (t
1/2for its 5.63 times), can also untoward reaction be alleviated while maintaining treatment effect.(3) AUC result of study display, the AUC (0-t) of T is equivalent to 3.51 times of R1 respectively, and point out this preparation curative effect high, thus consumption is little, has good potential applicability in clinical practice.
Contrast known, this preparation can slowly release norcantharidin in vivo, has good slow release characteristic, has good curative effect, and significantly reduces Cmax, reduces the risk and hidden danger brought because blood drug level is too high, has good potential applicability in clinical practice.
Claims (1)
1. a Norcantharidin derivative lipid microsphere injection, is characterized in that: this infusion pump contains following composition: norcantharidin imide N-alkyl derivative, oil phase, water and surfactant, and in the quality of the pharmaceutical preparations, recipe ingredient has:
Oil phase 5% ~ 30%,
In the norcantharidin imide N-alkyl derivative 0.001% ~ 5% of norcantharidin,
Surfactant 0.5% ~ 7%,
Osmotic pressure regulator 1% ~ 5%,
All the other are water for injection;
In described injection, norcantharidin imide N-alkyl derivative general structure is the compound of I;
Wherein: R is C for being selected from general formula
nh
2n+1straight chain saturated alkane base; N is an integer in 8-24.
2, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 1, is characterized in that: in described injection, osmotic pressure regulator is selected from one or more mixture in glycerol, sorbitol, mannitol, glucose.
3, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 1, is characterized in that: oil phase is selected from one or more mixture in mineral oil, vegetable oil, animal oil, quintessence oil and artificial oil; Surfactant is selected from phospholipid, tween, span, F68, one or more mixture in enuatrol, oleic acid, cholic acid, deoxycholic acid.
4, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 1 or 3, is characterized in that: described oil phase is selected from one or more mixture in soybean oil, safflower oil, Semen Maydis oil, MCT Oil, pearl barley oil, Petiolus Trachycarpi oil, Oleum Helianthi, Fructrs Hippophae seed oil, Radix Angelicae Sinensis oil, Fructus Zanthoxyli oil, Oleum Bulbus Allii, Oleum Curcumae or Rhizoma Chuanxiong oil.
5, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 3, is characterized in that: described tween is selected from the one in polysorbas20, polysorbate40, polysorbate60, Tween 80, polysorbate85; Described F68 is F68 F68; Described phospholipid is selected from fabaceous lecithin, lecithin or is both mixture.
6, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 1, is characterized in that: containing metal-chelator, antioxidant, buffer salt, pH adjusting agent in preparation; In the quality of the pharmaceutical preparations, metal-chelator is 0% ~ 1%, and antioxidant consumption is 0 ~ 2%, and pH adjusting agent consumption is 0.005% ~ 2%.
7, the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 6, is characterized in that: metal-chelator is one in disodium edetate, sodium calcium edetate or is both mixture; Antioxidant is one or more mixture in vitamin E, vitamin C, sodium sulfite, sodium pyrosulfite, cysteine; PH adjusting agent is the one in sodium hydroxide, hydrochloric acid.
8, a kind of preparation method of the lipide microsphere injection containing norcantharidin imide N-alkyl derivative as claimed in claim 1, it is characterized in that: by the quality of the pharmaceutical preparations of described claim 1, after norcantharidin imide N-alkyl derivative is mixed with oil phase, be redispersed in aqueous phase, prepare after colostrum through high-speed stirred, carry out high pressure homogenize process again, preparation adopts the sterilizing of high-pressure rotary sterilization.
9, the preparation method of the lipide microsphere injection containing norcantharidin imide N-alkyl derivative according to claim 8, is characterized in that: the method is undertaken by following processing step:
Injection osmotic pressure regulator, surfactant, metal-chelator and water soluble antioxidants are scattered in appropriate water for injection, put in magnetic stirring apparatus and be heated to 30-80
oc, and be stirred to whole dissolving, as aqueous phase;
By the norcantharidin imide N-alkyl derivative of recipe quantity and being dissolved in the oil phase of containing oil soluble antioxidant and oil phase solubilizing agent, put in magnetic stirring apparatus and be heated to 30-80
oc to mix homogeneously, as oil phase;
Under organizing at a high speed dispersion machine to stir, by aqueous phase and oil phase mixing, obtained colostrum;
Regulate pH value to about 4.0-9.0 colostrum sodium hydroxide or hydrochloric acid solution, be then settled to recipe quantity with water for injection, be transferred in high pressure homogenizer, control homogenizing temperature at 30-80 with ice-water bath
oc, to be about 20-80MPa pressure homogenizing 4-15 time;
Fill nitrogen, embedding, rotating type high-pressure steam sterilization, lowers the temperature rapidly, and after lamp inspection is qualified, packaging, in 4-10
oc stores.
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| CN102319214A (en) * | 2011-09-09 | 2012-01-18 | 沈阳药科大学 | Lipid microsphere injection containing sodium demethyl cantharidate-phosphatide complex and preparation method thereof |
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