CN103432601A - Perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent and preparation method thereof - Google Patents
Perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent and preparation method thereof Download PDFInfo
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- CN103432601A CN103432601A CN201310375354XA CN201310375354A CN103432601A CN 103432601 A CN103432601 A CN 103432601A CN 201310375354X A CN201310375354X A CN 201310375354XA CN 201310375354 A CN201310375354 A CN 201310375354A CN 103432601 A CN103432601 A CN 103432601A
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Abstract
The invention discloses a perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent and a preparation method thereof. The perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent refers to a perfluorooctylbromide coated mPEG-PLLA, PLGA-PEG-PLGA or mPEG-PCL block polymer ultrasound microbubble contrast agent. A preparation method of the agent comprises the steps of preparing an mPEG-PLLA, PLGA-PEG-PLGA or mPEG-PCL block polymer solution, adding perfluorooctylbromide perfluoropentane, perfluorohexane or perfluoropropane to the solution, and then preparing the ultrasound microbubble contrast agent by an electric internal cutting homogenizing, ultrasonic emulsifying or mechanical oscillating method. The perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent provided by the invention is applied to preparation of an ultrasonic imaging diagnosis contrast agent. The high polymer material used by the perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent is safe and nontoxic, and is obviously cheaper than synthetic phospholipids; the microbubble preparation process is simple and convenient; the novel perfluorooctylbromide coated block polymer ultrasound microbubble contrast agent has good potential prospect in clinical application.
Description
Technical field
The present invention relates to the ultrasound microbubble contrast agent that the fluorocarbons of liquid-gas phase transition can occur a kind of block polymer parcel, belong to the ultrasonic imaging diagnosis field.
Background technology
Ultrasound microbubble contrast agent has been widely used in the diagnosis of clinical disease, has obviously improved the ultrasonic diagnosis coincidence rate, is one " milestone " on the ultrasound medicine development history.The acoustic contrast agent of current clinical use mostly is lipid or albumin parcel noble gas, less stable, easily spontaneous breaking.
The advantages such as macromolecular material has good biocompatibility, biodegradable, adopt Polymer materialspreparation ultrasound microbubble contrast agent particle size distribution more even, and stability is high, and compressive property is good, has become the focus of current acoustic contrast agent research.But part macromolecular material, the contrast agent shell prepared as polylactic-co-glycolic acid is harder, the Second Harmonic Imaging ultrasonic contrast of low mechanical index is difficult to develop, need higher acoustics output just can cause the Non-Linear Vibration of microvesicle or break, produce the contrast imaging strengthened, but the background tissues echo signal also obviously strengthens simultaneously, causes signal to noise ratio obviously to reduce, the radiography weak effect.The present invention's research shows, macromolecular material is carried out to block polymer that the Polyethylene Glycol grafting process to form and as mono methoxy polyethylene glycol-Poly-L-lactic acid mPEG-PLLA, obviously improve the compliance of material, can form hydrophilic section PEG outwardly in aqueous solution, hydrophobic section PLLA micellar structure inwardly.
The perfluorocarbon compound of long fluorocarbon chain is in a liquid state when low temperature, i.e. liquid fluorocarbon, and when ambient pressure reduces or temperature while being elevated to its boiling point, the liquid fluorocarbon generating gasification, be transformed into gas.Perflenapent boiling point under normal pressure is 29 ℃, is expelled in human body and is gaseous state, is a kind of fluorocarbon material that is applicable to preparing liquid-gas phase transition type acoustic contrast agent.
Summary of the invention
The purpose of this invention is to provide a kind of block polymer ultrasound microbubble contrast agent that wraps up liquid fluorocarbon and preparation method thereof.
The present invention finds in the mPEG-PLLA block polymer ultrasound microbubble contrast agent of preparation parcel liquid fluorocarbon: change mPEG and PLLA molecular weight ratio, or change sheathing material into PEG-PCL mPEG-PCL, PEG-PLGA mPEG-PLGA, or reduce the homogenate of rotating speed (14500rpm) inscribe and carry out emulsifying, or change emulsification method into ultrasonic emulsification, or change liquid fluorocarbon into microvesicle density reduction prepared by sulfur hexafluoride, cause radiography to strengthen intensity and all obviously reduce.The ratio of the concentration of mPEG-PLLA aqueous solution, mPEG-PLLA and perflenapent etc. all can obviously affect output and the particle size distribution of microvesicle.
At first the present invention adopts dialysis to prepare the mPEG-PLLA micellar aqueous solution, under cryogenic conditions by liquid perflenapent and mPEG-PLLA aqueous solution, adopt high speed inscribe homogenate method to carry out emulsifying, successfully prepared and take block polymer as shell, the microbubble contrast agent of parcel perflenapent.After supposition may be liquid perflenapent and mPEG-PLLA aqueous solution, form the Emulsion of " oil-in-water " under high-speed homogenization, the heat that high-speed homogenization produces simultaneously makes the perflenapent generating gasification, and then the suitable microbubble contrast agent of formation particle diameter, particle size distribution is more even, stability obviously is better than the lipid microbubble contrast agent, is more suitable for carrying out ultrasonic targeting diagnosis and treatment research.
There is obvious reinforced effects under the Second Harmonic Imaging radiography pattern that the present invention adopts ultrasound microbubble contrast agent inside and outside prepared by single emulsion process to be 0.09 low mechanical index at MI, the microbubble contrast agent shell that has overcome the part Polymer materialspreparation is hard, is difficult to the shortcoming of developing.
The object of the present invention is achieved like this, the block polymer ultrasound microbubble contrast agent of described parcel liquid fluorocarbon, for mPEG-PLLA, the PLGA-PEG – PLGA of parcel liquid fluorocarbon or the ultrasound microbubble contrast agent of mPEG-PCL block polymer, it is made by following method: produce mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer solution, after adding liquid fluorocarbon perflenapent, perflexane or perfluoropropane, adopt electronic inscribe homogenate, ultrasonic emulsification or mechanical oscillation method to make ultrasound microbubble contrast agent.
The block polymer ultrasound microbubble contrast agent of described parcel perflenapent, in its block polymer: the mPEG molecular weight is that 1000-10000, PLLA molecular weight are that 2500-25000, PCL molecular weight are 2500-25000, and the block polymer solution concentration is 0.1-10 wt%; The ratio 10mg/1-100 μ l of block polymer and perflenapent.
The mPEG-PLLA block polymer ultrasound microbubble contrast agent of the parcel perflenapent that the electronic inscribe homogenate legal system of described preferred employing is standby, its homogenate rotating speed is 14500-35000rpm, homogenate time 0.5-4min.
The mPEG-PLLA block polymer ultrasound microbubble contrast agent of described preferred parcel perflenapent is made by following method: the mPEG-PLLA that to take molecular weight be 5000/5000 is dissolved in oxolane, the bag filter of packing into, spend the night with normal saline dialysis, change therebetween after dialysis solution and form the mPEG-PLLA normal saline solution; Get the mPEG-PLLA normal saline solution, the ice bath pre-cooling, add liquid perflenapent, and under condition of ice bath, electronic inscribe homogenate must be wrapped up the block polymer ultrasound microbubble contrast agent of perflenapent.
The mPEG-PLLA block polymer ultrasound microbubble contrast agent of described preferred parcel perflenapent is further made by following method: the mPEG-PLLA that to take the 0.2g molecular weight be 5000/5000 is dissolved in the 2ml oxolane, the bag filter of packing into, spend the night with normal saline dialysis, change dialysis solution therebetween and form the mPEG-PLLA normal saline solution 2 times; Get 1ml 1wt% mPEG-PLLA normal saline solution, the ice bath pre-cooling, add the liquid perflenapents of 40 μ l, and under condition of ice bath, the electronic inscribe homogenate of 35000rpm 2min, must wrap up the block polymer ultrasound microbubble contrast agent of perflenapent; The described microbubble contrast agent suspension that is creamy white,, observe under inverted microscope after described microbubble contrast agent dilution with normal saline, microvesicle is rounded, good dispersion, and smooth surface is bright, mean diameter is 3.7 ± 0.9 μ m, and adopting cell counting count board to measure microvesicle density is 2.3 * 10
8/ ml, 4 ℃ of amplitudes of placing the reduction of 24h microvesicle quantity are significantly lower than the lipid ultrasonic microbubble contrast agent.
The preparation method of the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon of the present invention, comprise the steps: to produce mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer solution, after adding liquid fluorocarbon perflenapent, perflexane or perfluoropropane, adopt electronic inscribe homogenate, ultrasonic emulsification or mechanical oscillation method to make ultrasound microbubble contrast agent.
In described mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer, the mPEG molecular weight is that 1000-10000, PLLA molecular weight are that 2500-25000, PCL molecular weight are 2500-25000, and the block polymer solution concentration is 0.1-10 wt%; The ratio 10mg/1-100 μ l of block polymer and perflenapent.
The mPEG-PLLA block polymer ultrasound microbubble contrast agent of the parcel perflenapent that the electronic inscribe homogenate legal system that the present invention preferably adopts is standby, its homogenate rotating speed is 14500-35000rpm, homogenate time 0.5-4min; Method for optimizing is: the mPEG-PLLA that to take molecular weight be 5000/5000 is dissolved in oxolane, and the bag filter of packing into spends the night and forms the mPEG-PLLA normal saline solution with normal saline dialysis; Get above-mentioned mPEG-PLLA normal saline solution, the ice bath pre-cooling, add liquid perflenapent, under condition of ice bath, must wrap up the block polymer ultrasound microbubble contrast agent of perflenapent after electronic inscribe homogenate.
The application of block polymer ultrasound microbubble contrast agent in preparing the ultrasonic imaging diagnosis contrast agent of parcel liquid fluorocarbon of the present invention.
In described application, after vein injects acoustic contrast agent, the microvesicle quantity of acoustic contrast agent is 4 * 10
7/ kg, carry out ultrasonic contrast and observe changes of echo, the TCA software analysis, and m-Strength Changes curve during drafting, finally can obtain Evaluation of Hepatic Arterial System after 3 seconds and start to strengthen, and and then liver parenchyma obviously strengthens, echo intensity peaking 6.92 * 10 in the time of 18 seconds
-5aU, continue the result disappear after 65 seconds or, after injecting acoustic contrast agent, carry out ultrasonic contrast and observe changes of echo, the TCA software analysis, m-Strength Changes curve during drafting, finally can obtain nephrolithotomy echo after several seconds and start to strengthen, to peak value, the result disappeared after continuing for some time.
The block polymer ultrasound microbubble contrast agent of parcel perflenapent of the present invention has following advantage: at first, adopt the macromolecular material of PEG grafting as the microvesicle shell, compliance is good, the liquid fluorocarbon gasification of parcel after expanding is supportted shell thin, make the low mechanical index ultrasonic energy under Second Harmonic Imaging radiography pattern pass the microvesicle shell, the obvious backscattering of the γ-ray emission of microvesicle inside, reach good ultrasonic contrast effect; Secondly, take block polymer as shell, can make it carry the medicines such as protein by polymer is modified, be a kind of novel medicine-carrying microvesicle contrast agent; Again, adopting the polymer of PEG grafting is shell, and good springiness improves mechanical index and carries out ultrasonic explosion and can make contrast agent destruction, can reach the microvesicle targeting and break and discharge the effect of medicine; Finally, the macromolecular material safety non-toxics such as mPEG-PLLA, the PLGA-PEG – PLGA that the present invention is used or mPEG-PCL block polymer, price is starkly lower than synthetic phospholipid, and microvesicle preparation technology is easy, be a kind of novel ultrasound microbubble contrast agent, there is potential potential applicability in clinical practice.
The accompanying drawing explanation
Fig. 1 is ultrasound microbubble contrast agent observation by light microscope figure of the present invention (* 400).
Fig. 2 is polymer microvesicle contrast agent and lipid microbubble contrast agent stability comparison diagram.
Fig. 3 is for getting 0.1ml microbubble contrast agent of the present invention, add the 20ml normal saline dilution, the water pocket of packing into, adopt Toshiba Aplio500 colorful Doppler ultrasound diagnostic apparatus to carry out ultrasonic contrast, under Second Harmonic Imaging ultrasonic contrast pattern, (MI=0.09) observes, in the water breakthrough capsule, echo obviously strengthens, and is intensive choice refreshments shape echogram.
Fig. 4 carries out ultrasonic explosion for improving mechanical index (MI=1.5), and in water pocket, microvesicle moment breaks, and echo reduction is echoless figure.
Fig. 5 is the timely m-intensity of ultrasound microbubble contrast agent rat liver radiography of the present invention.
Fig. 6 is the timely m-intensity of ultrasound microbubble contrast agent new zealand white rabbit nephrography of the present invention.
The specific embodiment
Below in conjunction with drawings and Examples, the present invention is described in detail.
Embodiment 1
1, the preparation of contrast agent
Taking 0.2g mPEG-PLLA(molecular weight is 5000/5000, is Mount Tai, Jinan handle of the Big Dipper biological engineering company limited product) be dissolved in the 2ml oxolane, the bag filter of packing into, spend the night to normal saline dialysis, changes dialysis solution therebetween 2 times; Get 1ml 1 wt % mPEG-PLLA normal saline solution, the ice bath pre-cooling, add 40 μ l perflenapents, and under condition of ice bath, the electronic inscribe homogenate of 35000rpm 2min, must wrap up the block polymer ultrasound microbubble contrast agent of perflenapent, and suspendible liquid is creamy white.Above-mentioned mPEG is mono methoxy polyethylene glycol, and PLLA is Poly-L-lactic acid.
2, the sign of contrast agent
With normal saline, by after the microbubble contrast agent dilution, under inverted microscope, observe, microvesicle is rounded, good dispersion, and smooth surface is bright, the more even (see figure 1) of size, mean diameter (3.7 ± 0.9) μ m, adopting cell counting count board to measure microvesicle density is 2.3 * 10
8/ ml.
3, stability experiment
Polymer microvesicle contrast agent and lipid microbubble contrast agent are placed to 4 ℃ of refrigerators, in 0,1,2,3,12,24h samples micro-Microscopic observation, lipid microbubble contrast agent quantity reduces amplitude apparently higher than polymer microvesicle contrast agent as a result, shows that polymer microvesicle contrast agent stability is apparently higher than lipid microbubble contrast agent (Fig. 2).
4, external radiography and explosion experiment
Get the microbubble contrast agent that 0.1ml step 1 makes, add the 20ml normal saline dilution, the water pocket of packing into, adopt Toshiba Aplio500 colorful Doppler ultrasound diagnostic apparatus to carry out ultrasonic contrast, under Second Harmonic Imaging ultrasonic contrast pattern, (MI=0.09) observes, in the water breakthrough capsule, echo obviously strengthens, and is intensive choice refreshments shape echo (see figure 3); Start burst mode (MI=1.5) microvesicle is broken up, in water pocket, echo moment reduction, be echoless (Fig. 4).Get the 20ml normal saline water pocket of packing into, contrasted, ultrasonic contrast is observed and is echoless.
5, rat liver radiography
Get female sd inbred rats, weight 180～220g, after the epigastrium depilation, ether inhalation anesthesia, adopt the tail intravenous bolus injection technique, injects the ultrasound microbubble contrast agent 4 * 10 of above-mentioned preparation
7/ kg, ultrasonic contrast is observed rat liver changes of echo, TCA software analysis, m-Strength Changes curve during drafting.As a result, after 3 seconds, the rats'liver Arterial system starts to strengthen, and and then liver parenchyma obviously strengthens, echo intensity peaking in the time of 18 seconds, and Power is 6.92 * 10
-5aU, continue approximately after 65 seconds, to disappear (Fig. 5).
6, new zealand white rabbit nephrography
Male new zealand white rabbit, weight 2.0～2.2 kg, be fixed in laboratory table, and auricular vein is inserted remaining needle, after the depilation of right side lower back, adopts vein group injecting method, injects ultrasound microbubble contrast agent 3 * 10
7/ kg, ultrasonic contrast is observed kidney changes of echo, TCA software analysis, m-Strength Changes curve during drafting.As a result, after 4 seconds, nephrolithotomy echo starts to strengthen, and peak value is 4.6 * 10
-4aU, continue approximately after 75 seconds, to disappear (Fig. 6).
Embodiment 2
Adopt the supersound method different from embodiment 1 to prepare the mPEG-PLLA contrast agent
Taking 0.2g mPEG-PLLA(molecular weight is 5000/5000, is Mount Tai, Jinan handle of the Big Dipper biological engineering company limited product) be dissolved in the 2ml oxolane, the bag filter of packing into, spend the night to normal saline dialysis, changes dialysis solution therebetween 2 times; Get 1ml 1 wt % mPEG-PLLA normal saline solution, the ice bath pre-cooling, add the liquid perflenapent of 40 μ l, under condition of ice bath, 200W ultrasonic emulsification 1min, result forms a large amount of large particle diameter microvesicles and the microcapsule mixed liquor that wraps up normal saline, and the microvesicle particle diameter is 6～50 μ m approximately, and microcapsule diameter is 3～7 μ m approximately.
Embodiment 3
The preparation of block polymer mPEG-PCL contrast agent
Taking 0.2g mPEG-PCL(molecular weight is 5000/5000, is Mount Tai, Jinan handle of the Big Dipper biological engineering company limited product, and PCL is polycaprolactone) be dissolved in the 2ml oxolane, the bag filter of packing into, spend the night to normal saline dialysis, changes dialysis solution therebetween 2 times; Get 1ml 1 wt% mPEG-PCL normal saline solution, the ice bath pre-cooling, add the liquid perflenapent of 40 μ l, under condition of ice bath, 200W ultrasonic emulsification 1min, obtain polymer microvesicle contrast agent, microvesicle mean diameter (4.5 ± 1.2) bigger than normal μ m, it is 1.2 * 10 that density is hanged down
8/ ml.
Embodiment 4
The preparation of polymer P LGA-PEG-PLGA contrast agent
Take 0.1g PLGA-PEG-PLGA(Poly(D,L-lactide-co-glycolide (PLGA) molecular weight 5000dal, PEG molecular weight 3000dal) be dissolved in the 10ml normal saline, get the 0.5ml aqueous solutions of polymers, add the liquid perflenapent of 20 μ l, mechanical oscillation 4000 times/minutes and 40 seconds, result forms a large amount of large particle diameter microvesicles and the microcapsule mixed liquor that wraps up normal saline, and the microvesicle particle diameter is 5～50 μ m approximately, and microcapsule diameter is 2～8 μ m approximately.
Sum up
The present invention can find out by four above-mentioned embodiment, 1) the microbubble contrast agent particle diameter of same block mPEG-PLLA polymer its formation under different preparation methoies, the uniformity, stability are all different, " under the condition of ice bath, the electronic inscribe homogenate of 35000rpm " that embodiment 1 adopts, microvesicle is rounded, good dispersion, smooth surface is bright, the more even (see figure 1) of size, mean diameter (3.7 ± 0.9) μ m, effect is better, 2) block polymer mPEG-PCL of the present invention, polymer P LGA-PEG-PLGA also can make corresponding ultrasound microbubble contrast agent under above-mentioned preparation method, but from the results of view, the block mPEG-PLLA polymer of the embodiment of the present invention 1 is at " its normal saline solution, in the ice bath pre-cooling, add perflenapent, under condition of ice bath, electronic inscribe homogenate, the block polymer ultrasound microbubble contrast agent of the parcel perflenapent obtained ", its microvesicle is rounded, good dispersion, smooth surface is bright, the more even (see figure 1) of size, mean diameter (3.7 ± 0.9) μ m, effect is best, 3) the present invention adopts liquid perfluorochemical, and the test display effect is best.
Specifically, 1) block polymer that the present invention adopts can be two blocks or triblock polymer, and mPEG/PLLA, PLGA/PEG, mPEG/PCL etc., wherein carry out with the mPEG-PLLA effect.MPEG-PLLA molecular weight ratio scope can be mPEG1000-10000, PLLA2500-25000, and it all can form ultrasound microbubble contrast agent, but best with the molecular weight proportioning effect of mPGE5000-PLLA5000.
2) preparation method of the present invention can adopt the methods such as electronic inscribe homogenate, ultrasonic emulsification, mechanical oscillation, wherein best with electronic inscribe homogenate effect, the microvesicle size is suitable, distribution uniform, homogenate rotating speed 14500-35000rpm, wherein best with the 35000rpm effect, homogenate time 0.5-4min, wherein the 2min effect is best.
3) the concentration 0.1-10 wt% of the above-mentioned mPEG-PLLA of the present invention, wherein with 1wt% effect best (wt% refers to weight percent concentration).
4) the present invention adopts fluorochemical as the gas in microvesicle, can select perflenapent, perfluoropropane, perflexane, sulfur hexafluoride etc., but best with the perflenapent effect, because it is in a liquid state when low temperature, it is liquid fluorocarbon, when ambient pressure reduces or temperature while being elevated to its boiling point, the liquid fluorocarbon generating gasification, and be transformed into more equably gas.
5) the ratio 10mg/1-100 μ l of mPEG-PLLA of the present invention and perflenapent, wherein best with 40 μ l effects.
Claims (10)
1. a block polymer ultrasound microbubble contrast agent that wraps up liquid fluorocarbon, for mPEG-PLLA, the PLGA-PEG – PLGA of parcel liquid fluorocarbon or the ultrasound microbubble contrast agent of mPEG-PCL block polymer, it is made by following method: produce mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer solution, after adding liquid fluorocarbon perflenapent, perflexane or perfluoropropane, adopt electronic inscribe homogenate, ultrasonic emulsification or mechanical oscillation method to make ultrasound microbubble contrast agent.
2. the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon according to claim 1, the block polymer ultrasound microbubble contrast agent that it is characterized in that described parcel perflenapent, in its block polymer: the mPEG molecular weight is that 1000-10000, PLLA molecular weight are that 2500-25000, PCL molecular weight are 2500-25000, and the block polymer solution concentration is 0.1-10 wt%; The ratio 10mg/1-100 μ l of block polymer and perflenapent.
3. the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon according to claim 1 and 2, the mPEG-PLLA block polymer ultrasound microbubble contrast agent that it is characterized in that the parcel perflenapent that the electronic inscribe homogenate of described employing legal system is standby, its homogenate rotating speed is 14500-35000rpm, homogenate time 0.5-4min.
4. the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon according to claim 3, the mPEG-PLLA block polymer ultrasound microbubble contrast agent that it is characterized in that described parcel perflenapent is made by following method: the mPEG-PLLA that to take molecular weight be 5000/5000 is dissolved in oxolane, the bag filter of packing into, spend the night with normal saline dialysis, change therebetween after dialysis solution and form the mPEG-PLLA normal saline solution; Get the mPEG-PLLA normal saline solution, the ice bath pre-cooling, add liquid perflenapent, and under condition of ice bath, electronic inscribe homogenate must be wrapped up the block polymer ultrasound microbubble contrast agent of perflenapent.
5. the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon according to claim 4, the mPEG-PLLA block polymer ultrasound microbubble contrast agent that it is characterized in that described parcel perflenapent is made by following method: the mPEG-PLLA that to take the 0.2g molecular weight be 5000/5000 is dissolved in the 2ml oxolane, the bag filter of packing into, spend the night with normal saline dialysis, change dialysis solution therebetween and form the mPEG-PLLA normal saline solution 2 times; Get 1ml 1wt% mPEG-PLLA normal saline solution, the ice bath pre-cooling, add the liquid perflenapents of 40 μ l, and under condition of ice bath, the electronic inscribe homogenate of 35000rpm 2min, must wrap up the block polymer ultrasound microbubble contrast agent of perflenapent; The described microbubble contrast agent suspension that is creamy white,, observe under inverted microscope after described microbubble contrast agent dilution with normal saline, microvesicle is rounded, good dispersion, and smooth surface is bright, mean diameter is 3.7 ± 0.9 μ m, and adopting cell counting count board to measure microvesicle density is 2.3 * 10
8/ ml, 4 ℃ of amplitudes of placing the reduction of 24h microvesicle quantity are significantly lower than the lipid ultrasonic microbubble contrast agent.
6. a preparation method of wrapping up the block polymer ultrasound microbubble contrast agent of liquid fluorocarbon, comprise the steps: to produce mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer solution, after adding liquid fluorocarbon perflenapent, perflexane or perfluoropropane, adopt electronic inscribe homogenate, ultrasonic emulsification or mechanical oscillation method to make ultrasound microbubble contrast agent.
7. the preparation method of the block polymer ultrasound microbubble contrast agent of parcel liquid fluorocarbon according to claim 6, it is characterized in that in mPEG-PLLA, PLGA-PEG – PLGA or mPEG-PCL block polymer, the mPEG molecular weight is that 1000-10000, PLLA molecular weight are that 2500-25000, PCL molecular weight are 2500-25000, the block polymer solution concentration is 0.1-10 wt%; The ratio 10mg/1-100 μ l of block polymer and perflenapent.
8. according to the preparation method of the block polymer ultrasound microbubble contrast agent of the described parcel liquid fluorocarbon of claim 6 or 7, it is characterized in that adopting the mPEG-PLLA block polymer ultrasound microbubble contrast agent of the standby parcel perflenapent of electronic inscribe homogenate legal system, its homogenate rotating speed is 14500-35000rpm, homogenate time 0.5-4min; Method for optimizing is: the mPEG-PLLA that to take molecular weight be 5000/5000 is dissolved in oxolane, and the bag filter of packing into spends the night and forms the mPEG-PLLA normal saline solution with normal saline dialysis; Get above-mentioned mPEG-PLLA normal saline solution, the ice bath pre-cooling, add liquid perflenapent, under condition of ice bath, must wrap up the block polymer ultrasound microbubble contrast agent of perflenapent after electronic inscribe homogenate.
9. the application of block polymer ultrasound microbubble contrast agent in preparing the ultrasonic imaging diagnosis contrast agent of the arbitrary described parcel liquid fluorocarbon of claim 1-5.
10. application according to claim 9, is characterized in that vein injects acoustic contrast agent, and the microvesicle quantity of acoustic contrast agent is 4 * 10
7/ kg, carry out ultrasonic contrast and observe changes of echo, the TCA software analysis, and m-Strength Changes curve during drafting, finally can obtain Evaluation of Hepatic Arterial System after 3 seconds and start to strengthen, and and then liver parenchyma obviously strengthens, echo intensity peaking 6.92 * 10 in the time of 18 seconds
-5aU, continue the result disappeared after 65 seconds or inject acoustic contrast agent, carries out ultrasonic contrast and observe changes of echo, the TCA software analysis, m-Strength Changes curve during drafting, finally can obtain nephrolithotomy echo after several seconds and start to strengthen, to peak value, the result disappeared after continuing for some time.
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| CN109453398A (en) * | 2018-09-05 | 2019-03-12 | 中山大学 | A kind of mesoporous poly-dopamine contains liquid fluorocarbon class acoustic contrast agent and preparation method thereof |
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