CN101756903A - Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome - Google Patents
Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome Download PDFInfo
- Publication number
- CN101756903A CN101756903A CN200910232406A CN200910232406A CN101756903A CN 101756903 A CN101756903 A CN 101756903A CN 200910232406 A CN200910232406 A CN 200910232406A CN 200910232406 A CN200910232406 A CN 200910232406A CN 101756903 A CN101756903 A CN 101756903A
- Authority
- CN
- China
- Prior art keywords
- carrying
- gene
- magnetic
- mnzn
- drug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a preparation method of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome, relating to the preparation process of a drug for treating cancer, in particular to nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome. Manganese sulfate, zinc sulfate and ferrisulphas are ground to powder in a certain ratio and are dissolved with a sodium hydroxide solution to obtain Mg-Zn-Fe magnetic nanoparticles by the steps of stirring, drying, roasting and the like; the obtained Mg-Zn-Fe magnetic nanoparticles are modified by a proper quantity of polyethyleneimine, and then modified Mg-Zn-Fe magnetic nanoparticles are combined with green fluorescent protein expression plasmid to obtain gene-carrying Mg-Zn-Fe magnetic nanoparticles; a proper quantity of dipalmitoyl phosphatidyl choline and cholesterol are added into an eggplant-shaped bottle to be dissolved by chloroform and absolute ether; then, the eggplant-shaped bottle is put into a rotary evaporator to be evaporated to cause the bottle bottom to form an even film; phosphate buffer containing the Mg-Zn-Fe magnetic gene-carrying nanoparticles and arsenic trioxide drug is added to be electrically stirred until the film falls off; and glycerol is added for repeated freeze thawing to obtain the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome.
Description
Technical field
The present invention relates to a kind of preparation technology as the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome for the treatment of cancer drug.
Background technology
That the present invention relates to is a kind of preparation technology of novel nano magnetic drug-carrying gene-carrying thermal controlled release liposome, and the even matter method of disperseing to add high pressure of the thin film by improvement prepares a kind of As that both had been encapsulated with
2O
3The MnZn ferromagnetic nano grain that is encapsulated with the carrying anti-tumor gene again has the novel nano liposome of temperature-controlled release of suitable phase transition temperature (41 ℃-46 ℃) simultaneously, because having magnetic induction in alternating magnetic field, the MnZn ferromagnetic nano grain of sealing heats up, temperature control, thermostatic characteristics, therefore when temperature rises to the phase transition temperature of liposome, medicine and gene then can slowly discharge, the chemotherapy under the thermal control of realization nano magnetic material is released and the combination of gene therapy, simultaneously, because the phase transition temperature of liposome of temperature-controlled release is 41 ℃-46 ℃, the ideal temperature of tumor thermotherapy just, thereby this novel nano magnetic drug-carrying gene-carrying thermal controlled release liposome can be treated with the thermal sensitive liposome for intermediary's thermal-arrest, chemotherapy, gene therapy is brought into play the good antitumor effect in one.Simultaneously, it also has advantages such as magnetic targeting location and reduction arsenical whole body toxic and side effects.
Thermal sensitive liposome is a kind of targeted drug carrier of rising in recent years, belongs to a kind of novel form of targeting drug delivery system, and it can utilize the medicine carrying characteristic of liposome and the double dominant of thermotherapy to improve therapeutic effect, reduces toxic and side effects.Its principle is when reaching phase transition temperature, phospholipid in the liposome produces and carries out the transition to mesomorphic physical transfer from colloidal state, the flowability of film and permeability increase, medicine is diffused in the target organ in a large number, tumor cell is on every side produced stronger lethal effect, thereby reach the effect of local chemotherapy,, can bring into play the effect of tumor thermotherapy simultaneously if phase transition temperature is between 41 ℃ to 46 ℃ of the tumor thermotherapy temperature.The scholar is arranged again in recent years
[1]Magnetic material such as iron dextran etc. is wrapped in liposome a kind of novel lipide-magnetic liposome that grows up, except possessing the characteristics of liposome, it can be under the effect of external magnetic field, the target area is optionally carried and be positioned to cancer therapy drug, thereby reduction dose, reduce toxicity, improve curative effect.The magnetic material main uses that is encapsulated in the magnetic liposome is the liposome magnetic target to tumor locus, improved the targeting of treatment, but the intensification thermotherapy function of tumor of nano magnetic material more and more comes into one's own in recent years, magnetic Nano material can strengthen thermotherapy effect in heat production under alternating magnetic field, and then improving the blood circulation in the tumor region and the permeability of blood vessel, the medicine that helps treating tumor reaches the cancer target zone.Also have the scholar directly to add cancer-resisting substance in magnetic Nano material, reach the double effects of thermotherapy and Drug therapy, magnetic material plays the dual function that strengthens heat production and targeted drug conveying in this process.Utilizing another outstanding advantage of magnetic Nano material is exactly that it can carry out temperature automatically controlled, constant temperature.Because ferromagnetic material itself has the characteristics of Curie temperature, ferromagnetism loses magnetism and lowers the temperature after temperature rises to curie point, be lower than and recover magnetic again after the curie point and heat up, thereby reach temperature automatically controlled and constant temperature to tumor thermotherapy, this thermotherapy to deep tumor has great importance.We successfully develop the magnetic manganese-zinc ferrite nano-particle with independent intellectual property right, and it is a kind of soft magnetic ferrite with low Curie temperature, can the strong absorption electromagnetic wave energy under the alternating magnetic field irradiation and heat up; When temperature reaches Curie temperature, it changes namagnetic substance into and loses the ability that absorbs electromagnetic wave energy, temperature is descended, when temperature is lower than Curie temperature, recover intensification manganese-zinc ferrite Nano microsphere not only but also recover the magnetic intensification, and so forth, make temperature be controlled at the Curie temperature place of setting all the time, thereby automatic intensification and temperature control that realization heats to treat tumor, the stability and the safety that improve therapeutic effect.We have also worked out when the x among the Mn1-xZnxFe2O4 is 0.5, and it can slowly rise to the suitable temperature (41 ℃-46 ℃) of oncotherapy and keep invariable after dispelling the heat through exchange, reaches the therapeutic effect of antitumor and don't damage normal structure.
The prospect of magnetic Nano material thermotherapy is very wide, if but simple thermotherapy does not have medicine carrying, and it is single that its effect still shows, and studies show that chemotherapeutics and combined with hyperthermia have significant synergism.
In the medicine of oncotherapy, arsenic and chemical compound thereof are the human tumors that is used for the treatment of the earliest, and arsenic trioxide injection is studied its effect to solid tumor both at home and abroad as national two kind new medicine official listings now, as breast carcinoma, pulmonary carcinoma, esophageal carcinoma, gastric cancer, confirmed its broad spectrum anticancer.In addition, As
2O
3When the clinical treatment leukemia, not only effect is remarkable, and the toxic and side effects such as bone marrow depression that do not have chemotherapeutics to cause, shortcomings such as the easy recurrence that no all-trans-retinoic acid treatment causes, easy drug resistance.But because systemic administration, very fast rising of blood plasma arsenic concentration and disperse surrounding tissue rapidly, thus still have the small part patient symptom of digestive tract, peripheral neuritis, xerosis cutis, pigmentation to occur, even side effect such as renal function injury appears.So, exploitation As
2O
3Novel form, the damage that the concentration of raising medicine in the target area reduces its hetero-organization is to improve the problem that antitumor curative effect urgently will solve.
In addition, along with the continuous development of Biochemistry and Molecular Biology, people have had further understanding to the molecule mechanism of various diseases.Thousands of kinds of diseases and gene-correlation are arranged, comprising ancestor genetic diseases relevant and the acquired disease relevant with polygenes with single-gene in known human diseases.The reason that causes disease mainly is genetic flaw and gene delection, suitable exogenous gene is introduced in the human body cell, with the gene that supplies a gap or lack, give expression to corresponding proteins matter, fundamentally eliminate the intrinsic factor that produce disease symptoms, and then produced a kind of new Therapeutic Method---gene therapy.Since the nineties in 20th century, as national great research project is classified human genome research one after another in countries in the world, along with the rapid progress of human genome research, people will reach a new level to the understanding on life gene basis, and also more and more for the research of gene importing carrier.In recent years, the extremely attention of researcher of carrier that liposome imports as gene, but it has natural degradation, non-immunogenicity, can repeat advantages such as transfection, be called as most promising gene transfection carrier in the field of gene, Vieweg etc. merge liposome and the plasmid that includes the IL-2 gene and form liposome-dna complex, it is hatched with prostate gland cancer cell, found that the IL-2 level obviously raises in the prostate gland cancer cell.Liposome portability range gene fragment; protecting group is not because of by nuclease degradation; and lipid and cell membrane merge behind the genes of interest transfered cell; lipid promptly is degraded; has unique advantages; therefore, liposome-mediated gene transfer method is first scheme that is applied to clinical gene therapy by American Cancer Society's approval.In addition, the polymine in the non-viral gene vector (PEI) also receives much attention, and it is a kind of organic macromolecule of high positive charge density that has, owing to contain a large amount of imido grpups in the molecule, PEI has very strong buffer capacity, the PH wide ranges.PEI can be fully protonated in the neutral water system, form " proton cotton balls ", thus can capture dna. scientist thinks that PEI also is the effective carrier of height of gene therapy, can carry out the transmission of former times acid of the few nuclear in inside and outside and plasmid.Therefore, the method for gene therapy by some specific modes and other treatment tumor being combined is a kind of pharmaceutical dosage form that has the treatment tumor of application prospect.
We at first modify back efficient capture Antioncogene with MnZn ferromagnetic nano material with PEI based on above achievement in research, adopt the thin film of the improvement even matter legal system of disperseing to add high pressure to be equipped with thermal sensitive liposome then, with As
2O
3Be encapsulated in wherein with the MnZn ferromagnetic nano grain of carrying anti-tumor gene, solve the phase transition temperature problem of liposome, be prepared into nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome, utilize the phase transition temperature (41 ℃-46 ℃) that manganese-zinc ferrite heats up automatically, temperature persistance reaches thermal sensitive liposome in alternating magnetic field, realize being encapsulated in the As in the liposome
2O
3With the control slow release of gene, reach thermochemotherapy, the common antineoplastic purpose of gene therapy, in addition, the magnetic material of sealing plays guiding and positioning action in liposome, make liposome adding under the action of a magnetic field, gather target organ, the targeting of treatment and specificity are strengthened with blood circulation.This research expectation by the gene therapy of liposome collection, thermotherapy, chemotherapy, thermal control release, advantage such as magnetic targeting is one, for tumor treatment has been opened up new thinking.
Summary of the invention
The invention provides a kind of preparation method of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome, it is strong to utilize the present invention to obtain magnetic responsiveness, particle diameter less (about 100nm), dispersibility are better, and envelop rate is higher, magnetic Nano material control slow release, liposome with gene therapy function.
The present invention adopts following technical scheme to solve its technical problem:
A kind of preparation method as the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome for the treatment of cancer drug:
The first step: with manganese sulfate MnSO
4H
2O, zinc sulfate ZnSO
47H
2O, ferrous sulfate FeSO
47H
2O mixing in 1: 1: 4 in molar ratio obtains compositions, place electric crusher to be ground into the end, also use dissolved in distilled water according to the metal ion in the described compositions and 1: 2.4 weighing sodium hydroxide of sodium hydroxide NaOH mol ratio then, again sodium hydrate aqueous solution is joined in the powder and and stirred three hours with electric blender, leave standstill in 80 ℃ of baking boxs of 12 hours postposition dry, put the interior 400 ℃ of roastings of Muffle furnace 1 hour again, after the cooling that the material porphyrize is powdered, embathe 6～8 times to remove solubility sodium sulfate inorganic salt with 50～60 ℃ of distilled water, use dehydrated alcohol drip washing afterwards, 60 ℃ of oven dry get MnZn ferromagnetism nanometer particle MZF-NP
Second step: be that the MnZn ferromagnetism nanometer particle is made into mass fraction is 4% magnetic fluid suspension for the oxalic acid solution of 5g/l with concentration, the centrifugal supernatant of abandoning behind the ultra-sonic dispersion, and the MnZn ferromagnetism nanometer particle after centrifugal joined in the phosphate buffer, ultrasonic limit, limit adds the abundant mixing of polymine PEI of 0.2 times of MnZn ferromagnetism nanometer particle quality, ultrasonic again 1 hour, magnetic agitation 2 hours, react fully and form stable polymine/MnZn Fe nanometer particles (PEI/MZF), use distilled water respectively, ethanol cyclic washing 6 times, after the drying polymine/MnZn Fe nanometer particles is fully mixed with egfp expression plasmid pEGFP, room temperature was placed 30 minutes, must carry gene MnZn ferromagnetism nanometer particle
The 3rd step: dipalmitoyl phosphatidyl choline DPPC mixes by mass ratio with cholesterol and places eggplant-shape bottle at 5: 3, and the chloroform of 10 times of volumes of lipid components and the mixed liquor of absolute ether in the adding dipalmitoyl phosphatidyl choline, chloroform and absolute ether volume ratio are 2: 1, it was dissolved fully in ultrasonic 3 minutes, with thin film rotary evaporator at 42 ℃ of following constant temperature rotary evaporation in vacuo, volatilize fully up to organic solvent, form a homogeneous film at the eggplant-shape bottle inwall; The gelatin that in phosphate buffer, adds arsenic trioxide As2O3 solution and 0.4～0.5 times of DPPC and cholesterol gross mass, As2O3 solution and phosphate buffer volume ratio are 1: 10, treat that in 60 ℃ ± 5 ℃ water-baths gelatin dissolving back adds year gene MnZn ferromagnetism nanometer particle of 1.5～2 times of DPPC and cholesterol gross mass, room temperature supersound process 3～5 minutes, form suspension, this suspension is added in the above-mentioned eggplant-shape bottle that contains thin film, electronic stirring is shaken to thin film and is come off, rotating speed is 2000 rev/mins, glycerol adding 1ml again, mixing, place-20 ℃ freezing 1～2 hour, 37 ℃ of thawings, supersound process place again after 30 seconds-20 ℃ 1～2 hour, so multigelation and supersound process promptly get nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome 3 times.
Compared with prior art, the present invention has obtained following beneficial effect:
1. adopt chemical coprecipitation successfully to prepare the manganese-zinc ferrite magnetic nano particle, use polyethylene imine beautify afterwards, size is 30～40nm under the polymine/MnZn Fe nanometer particles transmission electron microscope of preparation, favorable dispersibility, Fourier infrared spectrum analysis are seen the characteristic peak [with reference to Fig. 5] of polymine.Polymine/external magnetic induction the intensification of MnZn Fe nanometer particles ability is good, in frequency is that 200kHz, power are 4kW, the output heating current is under the alternating magnetic field irradiation of 300A, in 40 minutes, be raised to 45 ℃ of maximum temperatures, and the maintenance temperature constant, therefore can be used as a kind of magnetic induction heating nanoparticle and treat tumor.
2. the thin film of the present invention by the improvement even matter legal system of disperseing to add high pressure is equipped with more homogeneous of nano magnetic gene-loaded liposome of temperature-controlled release size, mean diameter is about 100nm, major part is single chamber [with reference to Fig. 1], magnetic responsiveness is good, the envelop rate height, stability is high, and phase transition temperature is 42.71 ℃, and the characteristic that its drug-carrying gene-carrying carries magnetic material all is better than the liposome with existing method preparation.
3. the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome of the present invention's preparation incorporates the effect of gene therapy first, because polymine is the effective carrier of height of gene therapy, the polymine of preparation/MnZn Fe nanometer particles energy efficient capture Antioncogene, confirmatory experiment shows, polymine/MnZn Fe nanometer particles can carry the pEGFP plasmid DNA and enter the HepG2 cell, make its expressing green fluorescent protein, behind polymine/MnZn Fe nanometer particles efficient capture Antioncogene, it is encapsulated in the liposome as gene transfer vector again, this method non-immunogenicity and cytotoxicity, can not cause the death of transformation and cell, and in the long time, organs such as liver function and renal function not had obvious toxic and side effects.
4. the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome of the present invention's preparation is compared with magnetic liposome in the past, most important character is that MnZn ferromagnetism nanometer particle that it is sealed by liposome self is warmed up to the suitableeest treatment temperature of tumor and lasting temperature control automatically and realizes that the thermal control of medicine and gene releases under the effect of external magnetic field, and it heats up phase transition temperature and magnetic material and has done more accurate combination.The performance of this foundation itself realizes the characteristic of the transformation of phase transition temperature, the control that can make the release of liposome release gene becomes simple and easy relatively, simultaneously, it is stronger that the effect of alternating magnetic field makes the thermal control of liposome release the controllability that becomes, realize oncotherapy regularly position, bonding part and permission gene and drug slow release, prolong action time effectively, and keep effective production concentration, improve partial antitumous effect.
5. certain amount of nano magnetic drug-carrying gene-carrying thermal controlled release liposome local injection is in transplanted tumor, in frequency is that 200kHz, power are that 4kW, output heating current are under the alternating magnetic field irradiation of 300A, the tumor area temperature is heated up about 45 ℃ rapidly, and keep constant temperature in the time afterwards, and the normal surrounding tissue temperature is constant substantially, show that this method has heating of targeting location and homothermic therapeutical effect, and the treatment of in-vivo tumour experimental results show that the effect of its combined chemotherapy, gene therapy is better than liposome dosage form in the past.
6. certain amount of nano magnetic drug-carrying gene-carrying thermal controlled release liposome interaction in vitro has very strong growth inhibited and apoptosis-promoting effect in human tumor cell line associating magnetic induction heating back pair cell, with simple bag medicine liposome, simple bag magnetic material liposome, reach simple gene therapy and compare, effect is (P＜0.05) more significantly.
7. nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome is that the treatment of a kind of energy thermal-arrest, chemotherapy, gene therapy are in the new antitumoral preparation of one, and have magnetic targeting, thermal control release, at multiple advantages such as treatment, side effect are little, be the antineoplastic new dosage form that has application prospect therefore.
Description of drawings
Fig. 1 is the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome that the present invention makes.
Fig. 2 includes manganese, zinc, ferrum, nitrogen, phosphorus, each element of arsenic and each component ratio under the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome power spectrum that makes of the present invention, and the success of novel form liposome preparation is described.
Fig. 3 is that polymine/MnZn Fe nanometer particles shifts the expression that the pEGFP plasmid DNA enters the green fluorescent protein (GFP) of HepG2 cell.
Fig. 4 is the heating curve of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome, illustrates that it has the stable also function of constant temperature control that heats up.
Fig. 5 is the Fourier infrared spectrum analysis, and the curve of top is the MnZn Fe nanometer particles, and lower curve is polymine/MnZn Fe nanometer particles, the characteristic peak of visible polymine.
The specific embodiment
A kind of preparation method of embodiment as the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome for the treatment of cancer drug:
A kind of preparation method as the nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome for the treatment of cancer drug:
The first step: with manganese sulfate MnSO
4H
2O, zinc sulfate ZnSO
47H
2O, ferrous sulfate FeSO
47H
2O mixing in 1: 1: 4 in molar ratio obtains compositions, place electric crusher to be ground into the end, also use dissolved in distilled water according to the metal ion in the described compositions and 1: 2.4 weighing sodium hydroxide of sodium hydroxide NaOH mol ratio then, again sodium hydrate aqueous solution is joined in the powder and and stirred three hours with electric blender, leave standstill in 80 ℃ of baking boxs of 12 hours postposition dry, put the interior 400 ℃ of roastings of Muffle furnace 1 hour again, after the cooling that the material porphyrize is powdered, embathe 6～8 times to remove solubility sodium sulfate inorganic salt with 50～60 ℃ of distilled water, use dehydrated alcohol drip washing afterwards, 60 ℃ of oven dry get MnZn ferromagnetism nanometer particle MZF-NP
Second step: be that the MnZn ferromagnetism nanometer particle is made into mass fraction is 4% magnetic fluid suspension for the oxalic acid solution of 5g/l with concentration, the centrifugal supernatant of abandoning behind the ultra-sonic dispersion, and the MnZn ferromagnetism nanometer particle after centrifugal joined in the phosphate buffer, ultrasonic limit, limit adds the abundant mixing of polymine PEI of 0.2 times of MnZn ferromagnetism nanometer particle quality, ultrasonic again 1 hour, magnetic agitation 2 hours, react fully and form stable polymine/MnZn Fe nanometer particles (PEI/MZF), use distilled water respectively, ethanol cyclic washing 6 times, after the drying polymine/MnZn Fe nanometer particles is fully mixed with egfp expression plasmid pEGFP, room temperature was placed 30 minutes, must carry gene MnZn ferromagnetism nanometer particle
The 3rd step: dipalmitoyl phosphatidyl choline DPPC mixes by mass ratio with cholesterol and places eggplant-shape bottle at 5: 3, and the chloroform of 10 times of volumes of lipid components and the mixed liquor of absolute ether in the adding dipalmitoyl phosphatidyl choline, chloroform and absolute ether volume ratio are 2: 1, it was dissolved fully in ultrasonic 3 minutes, with thin film rotary evaporator at 42 ℃ of following constant temperature rotary evaporation in vacuo, volatilize fully up to organic solvent, form a homogeneous film at the eggplant-shape bottle inwall; The gelatin that in phosphate buffer, adds arsenic trioxide As2O3 solution and 0.4～0.5 times of DPPC and cholesterol gross mass, As2O3 solution and phosphate buffer volume ratio are 1: 10, treat that in 60 ℃ ± 5 ℃ water-baths gelatin dissolving back adds year gene MnZn ferromagnetism nanometer particle of 1.5～2 times of DPPC and cholesterol gross mass, room temperature supersound process 3～5 minutes, form suspension, this suspension is added in the above-mentioned eggplant-shape bottle that contains thin film, electronic stirring is shaken to thin film and is come off, rotating speed is 2000 rev/mins, glycerol adding 1ml again, mixing, place-20 ℃ freezing 1～2 hour, 37 ℃ of thawings, supersound process place again after 30 seconds-20 ℃ 1～2 hour, so multigelation and supersound process promptly get nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome 3 times.
Claims (1)
1. preparation method of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome as the treatment cancer drug is characterized in that:
The first step: with manganese sulfate MnSO
4H
2O, zinc sulfate ZnSO
47H
2O, ferrous sulfate FeSO
47H
2O mixing in 1: 1: 4 in molar ratio obtains compositions, place electric crusher to be ground into the end, also use dissolved in distilled water according to the metal ion in the described compositions and 1: 2.4 weighing sodium hydroxide of sodium hydroxide NaOH mol ratio then, again sodium hydrate aqueous solution is joined in the powder and and stirred three hours with electric blender, leave standstill in 80 ℃ of baking boxs of 12 hours postposition dry, put the interior 400 ℃ of roastings of Muffle furnace 1 hour again, after the cooling that the material porphyrize is powdered, embathe 6～8 times to remove solubility sodium sulfate inorganic salt with 50～60 ℃ of distilled water, use dehydrated alcohol drip washing afterwards, 60 ℃ of oven dry get MnZn ferromagnetism nanometer particle MZF-NP
Second step: be that the MnZn ferromagnetism nanometer particle is made into mass fraction is 4% magnetic fluid suspension for the oxalic acid solution of 5g/l with concentration, the centrifugal supernatant of abandoning behind the ultra-sonic dispersion, and the MnZn ferromagnetism nanometer particle after centrifugal joined in the phosphate buffer, ultrasonic limit, limit adds the abundant mixing of polymine PEI of 0.2 times of MnZn ferromagnetism nanometer particle quality, ultrasonic again 1 hour, magnetic agitation 2 hours, react fully and form stable polymine/MnZn Fe nanometer particles (PEI/MZF), use distilled water respectively, ethanol cyclic washing 6 times, after the drying polymine/MnZn Fe nanometer particles is fully mixed with egfp expression plasmid pEGFP, room temperature was placed 30 minutes, must carry gene MnZn ferromagnetism nanometer particle
The 3rd step: dipalmitoyl phosphatidyl choline DPPC mixes by mass ratio with cholesterol and places eggplant-shape bottle at 5: 3, and the chloroform of 10 times of volumes of lipid components and the mixed liquor of absolute ether in the adding dipalmitoyl phosphatidyl choline, chloroform and absolute ether volume ratio are 2: 1, it was dissolved fully in ultrasonic 3 minutes, with thin film rotary evaporator at 42 ℃ of following constant temperature rotary evaporation in vacuo, volatilize fully up to organic solvent, form a homogeneous film at the eggplant-shape bottle inwall; The gelatin that in phosphate buffer, adds arsenic trioxide As2O3 solution and 0.4～0.5 times of DPPC and cholesterol gross mass, As2O3 solution and phosphate buffer volume ratio are 1: 10, treat that in 60 ℃ ± 5 ℃ water-baths gelatin dissolving back adds year gene MnZn ferromagnetism nanometer particle of 1.5～2 times of DPPC and cholesterol gross mass, room temperature supersound process 3～5 minutes, form suspension, this suspension is added in the above-mentioned eggplant-shape bottle that contains thin film, electronic stirring is shaken to thin film and is come off, rotating speed is 2000 rev/mins, glycerol adding 1ml again, mixing, place-20 ℃ freezing 1～2 hour, 37 ℃ of thawings, supersound process place again after 30 seconds-20 ℃ 1～2 hour, so multigelation and supersound process promptly get nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome 3 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910232406A CN101756903A (en) | 2009-11-27 | 2009-11-27 | Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910232406A CN101756903A (en) | 2009-11-27 | 2009-11-27 | Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101756903A true CN101756903A (en) | 2010-06-30 |
Family
ID=42488389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910232406A Pending CN101756903A (en) | 2009-11-27 | 2009-11-27 | Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101756903A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225209A (en) * | 2011-06-26 | 2011-10-26 | 东南大学 | Preparation method of nano magnetic granule composite system |
| CN102286531A (en) * | 2011-06-26 | 2011-12-21 | 东南大学 | Use of mangan zinc ferrite nano magnetic particles modified by polyethylenimine (PEI) |
| CN103989634A (en) * | 2014-05-07 | 2014-08-20 | 东南大学 | Heat controlled release nano-magnetic arsenic liposome and preparation method thereof |
| CN106668882A (en) * | 2017-03-22 | 2017-05-17 | 郑州大学 | Ultrasound-sensitive liposome and application thereof |
| CN112386694A (en) * | 2019-08-12 | 2021-02-23 | 湖南早晨纳米机器人有限公司 | Magnesium alloy thermal therapy nano robot and preparation method thereof |
| CN112386691A (en) * | 2019-08-12 | 2021-02-23 | 湖南早晨纳米机器人有限公司 | Preparation method of far infrared drug-loaded nano robot |
| WO2022183679A1 (en) * | 2021-03-03 | 2022-09-09 | 张铿 | Transdermal drug delivery layer |
-
2009
- 2009-11-27 CN CN200910232406A patent/CN101756903A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225209A (en) * | 2011-06-26 | 2011-10-26 | 东南大学 | Preparation method of nano magnetic granule composite system |
| CN102286531A (en) * | 2011-06-26 | 2011-12-21 | 东南大学 | Use of mangan zinc ferrite nano magnetic particles modified by polyethylenimine (PEI) |
| CN103989634A (en) * | 2014-05-07 | 2014-08-20 | 东南大学 | Heat controlled release nano-magnetic arsenic liposome and preparation method thereof |
| CN106668882A (en) * | 2017-03-22 | 2017-05-17 | 郑州大学 | Ultrasound-sensitive liposome and application thereof |
| CN112386694A (en) * | 2019-08-12 | 2021-02-23 | 湖南早晨纳米机器人有限公司 | Magnesium alloy thermal therapy nano robot and preparation method thereof |
| CN112386691A (en) * | 2019-08-12 | 2021-02-23 | 湖南早晨纳米机器人有限公司 | Preparation method of far infrared drug-loaded nano robot |
| WO2022183679A1 (en) * | 2021-03-03 | 2022-09-09 | 张铿 | Transdermal drug delivery layer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101756903A (en) | Preparation process of nanometer magnetic drug-carrying gene-carrying thermal controlled release liposome | |
| CN104623658B (en) | A kind of water-soluble ferrite composite nanoparticle and its preparation method and application | |
| CN101744766A (en) | Preparing technique of nano magnetic gene-loaded liposome of temperature-controlled release | |
| CN109091674B (en) | Multifunctional drug carrier and preparation method and application thereof | |
| CN107335063A (en) | A kind of Anti-cancer biologic film nano target is with plastid and preparation method and application | |
| CN111603559B (en) | Copper iodophor compound @ photosensitizer composite nanoparticle and application thereof as X-ray photodynamic therapy medicine | |
| Zhao et al. | Immunostimulatory multi-interfacial bimetallic phosphide nanoparticles as photo-enhanced cascade nanozyme for cancer therapy | |
| CN103611172A (en) | Nanorealgar-carrying magnetic albumin nanospheres and preparation method thereof | |
| CN114177286A (en) | Injectable self-healing hydrogel encapsulating mesoporous nano microspheres and preparation method and application thereof | |
| CN111759808B (en) | Liposome-graphene-gold composite nano material and preparation method and application thereof | |
| CN113616811A (en) | Fusion type multifunctional nano vesicle modified by apolipoprotein as well as preparation method and application thereof | |
| CN109550050B (en) | Melanin-loaded molybdenum dioxide drug-loaded compound and preparation and application thereof | |
| CN111286326B (en) | Preparation method and application of silicate long-afterglow probe | |
| CN105663043A (en) | 131I-labelled c(RGDyk)-targeting doxorubicin thermosensitive nanoliposome and preparation and application thereof | |
| Zhang et al. | Ultrasound-enhanced cascade chemodynamic tumor nanotherapy with lactic acid-enabled hydrogen peroxide self-production | |
| CN106334190A (en) | Compound drug carrier for multi-response mechanism and preparation method thereof | |
| CN113289014B (en) | Zwitterionic polypeptide/gallic acid/iron coordination nanoparticle with ultra-small particle size as well as preparation method and application thereof | |
| CN1242820C (en) | Nano magnetic heating seed for thermotherapy, its preparation method and use | |
| CN110917172B (en) | Molybdenum oxide nanosheet plugging hollow mesoporous silicon nanomaterial and preparation and application thereof | |
| CN115040643A (en) | Tumor cell-bacterium fusion material and preparation method and application thereof | |
| CN101987203A (en) | Method for preparing magnetic gene-loaded lipid ultrasonic microbubble contrast medium | |
| CN111265495A (en) | Copper sulfide and ovalbumin-loaded composite nanoparticle and preparation method thereof | |
| CN103989634A (en) | Heat controlled release nano-magnetic arsenic liposome and preparation method thereof | |
| CN1241550C (en) | Method for preparing nanometer white arsenic magnetic liposome | |
| CN109675056B (en) | Controlled release system based on base pairing rule and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20100630 |