CN101062423A - Gas emulsion type Ultrasound Contrast Agent microspheres and the preparing method thereof - Google Patents
Gas emulsion type Ultrasound Contrast Agent microspheres and the preparing method thereof Download PDFInfo
- Publication number
- CN101062423A CN101062423A CN 200710106559 CN200710106559A CN101062423A CN 101062423 A CN101062423 A CN 101062423A CN 200710106559 CN200710106559 CN 200710106559 CN 200710106559 A CN200710106559 A CN 200710106559A CN 101062423 A CN101062423 A CN 101062423A
- Authority
- CN
- China
- Prior art keywords
- contrast agent
- gas
- microsphere
- acoustic contrast
- phospholipid
- 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
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a new type gas emulsion supersonic contrast agent and preparing method, which is characterized by the following: choosing phosphatide, carbowax derivant and Bolusham as raw material; choosing the coating gas as water-insoluble gas with bioavailability; adding the raw material into equal liquid penetration; dispersing completely; proceeding supersonic vibrate or mechanical cutting; getting the product. This invention possesses low cost, simple preparation and good stability.
Description
Invention field
The present invention relates to a kind of acoustic contrast agent, be specially by phospholipid, a kind of aerial emulsion type ultraphonic contrast agent microballoon that polyethyleneglycol derivative and poloxamer etc. form as ball wall fraction parcel fluorine carbon insoluble gas, and relate to its preparation method.
Background of invention
With clinical other medical diagnostic techniqu relatively, ultrasonic have advantages such as radiationless and cheap.When carrying out the ultrasonic image inspection, Ultrasound Instrument emission ultrasound wave, the acoustic signals that receives organ organizational structure reflected back again carries out the image that data conversion forms the examine zone.
The reflection of sound wave at different acoustic densities interface (as liquid-gas) is very effective.Therefore select for use suitable ball wall material parcel gas to make the gassiness microsphere and when ultrasonic imaging diagnosis, can be used as the acoustic contrast agent use, can realize that blood flow and histoorgan echo strengthen after being injected into human body, improve histoorgan ultrasonogram quality greatly, and can help to understand the blood perfusion situation of organ, tissue.
The research of relevant acoustic contrast agent is always among constantly improving, ideal acoustic contrast agent should possess following characteristic: 1) the suitable particle diameter of tool, on the one hand, microspherulite diameter is big more, the ultrasonic reflection ability is strong more, human capillary vessel's mean inside diameter is less than 8 μ m on the other hand, and therefore for making contrast agent microsphere free-flow not disturb hemodynamics in blood capillary, microsphere diameter should be controlled within the specific limits.2) the enough concentration of tool makes ultrasonic reflection signal enough strong.3) good stability, the effect that can tolerate human body artery pressure (about 250mmHg) is not destroyed, and keeps the ultrasonic reflection function.4) circulation time is long in vivo, and safety is good.
For making ideal acoustic contrast agent, up-to-date technology concentrates in the selection to the improvement of ball wall material and process.
For example, it is the gassiness microsphere suspension of ball wall material with the albumin that Chinese patent CN1081467C discloses, and preparation method is a ultrasonic method.Chinese patent CN1128638C discloses the class with surfactant department, and the microsphere suspension that parcel gases such as tween form makes with ultrasonic method.Chinese patent CN1209167C discloses by lipid and Biodegradable high-molecular polymer etc. and has been the contrast agent microsphere of filmogen, gets with the emulsion polymerization preparation.
In the existing contrast agent, much still be in each stage of research and development.Albumin microsphere is used early, existing procucts exploitation listing, and albumin microsphere ball wall shell is a heat denatured protein, and this has caused the elasticity (being compressibility) of microsphere to reduce, and therefore retention time is shorter in vivo, has limited its application to a certain extent.Consider class of department, the general less intravenous formulations that is used for of Tweens surfactant from the safety aspect; And owing to often will use organic solvent in preparation technology, can there be the organic solvent residual problem in the microsphere of pbz polymer polymer.In fact,, still need the ball wall is further studied improvement, carefully select material and determine ratio, with the contrast agent product that more high-quality, safety is provided and is easy to prepare for obtaining acoustic contrast agent safely and effectively.
Summary of the invention
Purpose of the present invention is promptly to provide that a kind of particle diameter is suitable, good stability, the effectively acoustic contrast agent and the rational preparation method of the long application safety of developing time.
Aerial emulsion type ultraphonic contrast agent ball wall material of the present invention comprises phospholipid, polyethyleneglycol derivative, poloxamer.In the described contrast agent, content of phospholipid is 0.02～0.5 weight %, and polyethyleneglycol derivative content is 0.001～0.04 weight %, and poloxamer content is 0.004～0.2 weight %, and the gas that is wrapped in the microsphere is fluorocarbon gas, and solvent is an isotonic solution.
Above-mentioned phospholipid is selected from lecithin, soybean phospholipid, and soybean lecithin, phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE, phosphatidic acid, Phosphatidylserine and phosphatidyl glycerol, above-mentioned phospholipid are human body n cell film component, and be nontoxic, non-immunogenicity.Wherein, preferably select lecithin for use, raw material is easy to get, and cost is low, and content is the best with 0.05～0.15 weight %.
Above-mentioned polyethyleneglycol derivative comprises Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE (PEG-PE), and Polyethylene Glycol-phosphatidylcholine (PEG-PC) is preferably selected the PEG4000-DSPE for use.Polyethyleneglycol derivative content is the best with 0.003～0.008 weight %.The adding of Polyethylene Glycol, microsphere surface is covered by submissive and hydrophilic polyethylene glycol long chain, can stop effectively in the blood many different components particularly opsonin to the absorption of microsphere, thereby reduced the affinity of mononuclear phagocyte, prolonged the residence time of microsphere in blood circulation microsphere.
Above-mentioned poloxamer content is the best with 0.015～0.04 weight %.The adding of poloxamer helps the formation of gas Emulsion microsphere and stable.
Be wrapped in gas in the microsphere and in contrast agent, bring into play the effect of ultrasonic reflection layer.Described gas is biocompatible water-insoluble gas, preferred perfluoropropane gas.The perfluoropropane boiling point is low, is gas at normal temperatures, and is water insoluble, is difficult for overflowing from microsphere, can keep the integrity of microsphere in aqueous medium.Perfluoropropane is nontoxic, be biological noble gas, but Secure Application is in human body.
Above-mentioned solvent is an isotonic solution, is selected from 5% Glucose Liquid, 2.5% glycerin liquid, 5% sorbitol liquid and 5% xylitol liquid.Inject in the body being suitable for.
The preparation method of above-mentioned acoustic contrast agent of the present invention is as follows:
Step a) takes by weighing in the vadose solution agent such as phospholipid, polyethyleneglycol derivative, poloxamer add under 50-60 ℃ of condition fully mixing by above-mentioned formula ratio.
Step b) feeds the described solution of step a with fluorocarbon gas.
Step c)
1) mixture of using ultrasound ripple processor sonic oscillation Treatment Solution and gas makes emulsifying mixture, the microsphere suspension of the parcel gas that the formation particle diameter meets the requirements.
Or 2) mixture of application shearing equipment Treatment Solution and gas makes emulsifying mixture by the mechanical shear stress effect, the microsphere suspension of the parcel gas that the formation particle diameter meets the requirements.
Carry out lyophilization in the microsphere suspension immigration freeze dryer of step d) with step C gained, make dried frozen aquatic products.
Before step e) is faced usefulness, add sterile water for injection, dried frozen aquatic products is redissolved, promptly get the microsphere aqueous suspension.
Preparation method of the present invention, be that fluorine carbon gas is introduced in the solution, under sonic oscillation or mechanical shear stress effect, phospholipid, polyethyleneglycol derivative, poloxamer are made aerial emulsion type ultraphonic contrast agent microballoon at liquid-vapor interface formation of deposits ball wall shell with the gas parcel.The present invention adopts sonic oscillation and mechanical shearing method to prepare the less investment of contrast agent microsphere equipment, operates simple and easy.It is method for optimizing that the mechanical shearing of wherein using homogenizer or colloid mill is used for preparing contrast agent microsphere, compare with ultrasonic method, the easier technology of carrying out is amplified to be used for enlarging and is produced, it is this method major parameter that the shearing of equipment disperses rotating speed, can regulate as required to prepare concentration, the suitable contrast agent product of particle diameter.
Describe the present invention in detail below in conjunction with embodiment.
The specific embodiment
Embodiment 1: the influence that poloxamer forms microsphere
Take by weighing lecithin by 0.1 weight %, 0.005 weight % takes by weighing the PEG4000-DSPE, respectively with 0 weight %, 0.015 weight %, 0.025 weight %, 0.035 the poloxamer of the weight % amount of taking by weighing mixes, add in 5% Glucose Liquid under 50-60 ℃ of condition fully mixing, feed perfluoropropane gas, disperse cutter head to stretch into homogenizer and carry out the high speed shear dispersion under the liquid level, make milky contrast agent microsphere suspension,, the results are shown in Table 1 with Ku Erte grain graininess analysis-e/or determining microsphere concentration and particle diameter.
The influence that table 1 poloxamer content forms microsphere
| Poloxamer concentration (weight %) | Microsphere concentration (10 8/ml) | Microsphere average diameter (μ m) |
| 0 | 4.3 | 3.6 |
| 0.015 | 8.92 | 3.0 |
| 0.025 | 8.58 | 2.9 |
| 0.035 | 9.87 | 2.7 |
The result shows, the adding of Sq poloxamer helps the formation of gas Emulsion microsphere, generates the microsphere suspension of high concentration.
Embodiment 2: the equipment rotating speed is to the influence of microsphere productive rate and particle diameter
Take by weighing lecithin by 0.1 weight %, 0.005 weight % takes by weighing the PEG4000-DSPE, 0.025 weight % takes by weighing poloxamer, add in 5% Glucose Liquid under 50-60 ℃ of condition fully mixing, feed perfluoropropane gas, disperse cutter head to stretch under the liquid level homogenizer, with different rotating speeds solution is carried out high speed shear and disperse, the concentration and the particle size data that make microsphere see Table 2.
Table 2 production technology is to the influence of microsphere concentration and particle diameter
| Rotating speed (r/min) | Microsphere concentration (10 8/ml) | Microsphere average diameter (μ m) |
| 17500 | 7.39 | 3.8 |
| 21500 | 9.05 | 3.4 |
| 24000 | 10.92 | 2.9 |
As can be seen from Table 2, along with the increase that disperses the cutter head rotating speed, the microsphere concentration that makes increases and mean diameter reduces.
Embodiment 3: the test of microsphere resistance to pressure
Take by weighing lecithin by 0.1 weight %, 0.005 weight % takes by weighing the PEG4000-DSPE, 0.025 weight % takes by weighing poloxamer, make the contrast agent microsphere suspension according to embodiment 1 described method with the rotating speed of 24000r/min and be sub-packed in ampoule, the application of pressure device applies with 100mmHg suspension respectively, 200mmHg, the pressure of 300mmHg 2 minutes, measure the concentration and the particle diameter of microsphere then with kurt instrument, the data before and after the pressurization relatively see Table 3.
Table 3 air pressure is to the influence of microsphere concentration and particle diameter
| Proof pressure (mmHg) | Microsphere concentration (10 8/ml) | Microsphere average diameter (μ m) |
| 0 | 1.45 | 2.83 |
| 100 | 1.44 | 2.80 |
| 200 | 1.46 | 2.78 |
| 300 | 1.42 | 2.76 |
As can be seen from Table 3, Ultrasound Contrast Agent microspheres resistance to pressure of the present invention is good, can tolerate the effect of human body artery pressure after the intravenous injection fully, realizes that ultrasonic development contrasts enhanced effect.
Embodiment 4: acoustic contrast agent in-vivo imaging experiment of the present invention
Behind experimental dog peripheral intravenous injection acoustic contrast agent of the present invention, kidney of dog (Fig. 1) and heart (Fig. 2) are all obtained satisfied acoustics video picture and are strengthened.Prompting, this contrast agent can be used for the perfused tissue imaging.Inject acoustic contrast agent of the present invention and do not influence experimental dog heart rate, blood pressure (table 4).
Table 4 experimental dog is at the injection forward and backward heart rate of acoustic contrast agent of the present invention and blood pressure situation (x ± s)
| Blood pressure (mmHg) | Heart rate (inferior/minute) | ||
| Systolic pressure | Diastolic pressure | ||
| Experimental example number (N) behind the ultrasonic contrast before the ultrasonic contrast | 100.2±7.8 105.2±7.3 6 | 67.7±15.8 79.8±7.6 6 | 133.7±10.5 124±11.1 6 |
| t p | -1.987 0.104 | -1.411 0.217 | 1.27 0.26 |
Description of drawings
Fig. 1, dog kidney acoustic contrast development effect figure.Left side figure is a kidney background image before the injection of contrast medium; After right figure is injection of contrast medium, experimental dog renal perfusion developed image.
Fig. 2, dog cardiac acoustic contrast development effect figure.Left side figure is the background image of experimental dog heart before the injection of contrast medium; Right figure is an experimental dog heart perfusion developed image behind the injection of contrast medium.
Claims (14)
1, a kind of aerial emulsion type ultraphonic contrast agent is the microsphere class aqueous suspension of 2～5 microns parcel gas for average diameter, and it is characterized in that: the ball wall material comprises phospholipid, polyethyleneglycol derivative, poloxamer; In the described contrast agent, content of phospholipid is 0.02～0.5 weight %, and polyethyleneglycol derivative content is 0.001～0.04 weight %, and poloxamer content is 0.004～0.2 weight %, the gas that is wrapped in the microsphere is biocompatible water-insoluble gas, and solvent is an isotonic solution.
2, according to the acoustic contrast agent of claim 1, wherein said phospholipid is selected from lecithin, soybean phospholipid, soybean lecithin, phosphatidylcholine, PHOSPHATIDYL ETHANOLAMINE, phosphatidic acid, Phosphatidylserine and phosphatidyl glycerol.
3, according to the acoustic contrast agent of claim 2, wherein said phospholipid is lecithin.
4, according to the acoustic contrast agent of claim 1 or 2 or 3, wherein said content of phospholipid is 0.05～0.15 weight %.
5, according to the acoustic contrast agent of claim 1, wherein polyethyleneglycol derivative comprises Polyethylene Glycol-PHOSPHATIDYL ETHANOLAMINE (PEG-PE), Polyethylene Glycol-phosphatidylcholine (PEG-PC).
6, according to the acoustic contrast agent of claim 5, wherein said polyethyleneglycol derivative is selected from the PEG4000-DSPE.
7, according to the acoustic contrast agent of claim 1 or 5 or 6, wherein said polyethyleneglycol derivative content is 0.003～0.008 weight %.
8, according to the acoustic contrast agent of claim 1, wherein said poloxamer content is 0.015～0.04 weight %.
9, according to the acoustic contrast agent of claim 1, the dissolubility during 25 ℃ of wherein said gases in water is to be less than 0.01ml gas in every ml water.
10, according to the acoustic contrast agent of claim 9, wherein said gas is perfluoropropane.
11, according to the acoustic contrast agent of claim 1, wherein said solvent is selected from 5% Glucose Liquid, 2.5% glycerin liquid, 5% sorbitol liquid and 5% xylitol liquid.
12, prepare the method for the arbitrary acoustic contrast agent of claim 1-11, it comprises:
Step a) with in the vadose solution agent such as phospholipid, polyethyleneglycol derivative, poloxamer add under 50-60 ℃ of condition abundant mixing.
Step b) feeds the described solution of step a with fluorocarbon gas.
Step c)
1) mixture of using ultrasound ripple processor sonic oscillation Treatment Solution and gas makes emulsifying mixture, the microsphere suspension of the parcel gas that the formation particle diameter meets the requirements.
Or 2) mixture of application shearing equipment Treatment Solution and gas makes emulsifying mixture by the mechanical shear stress effect, the microsphere suspension of the parcel gas that the formation particle diameter meets the requirements.
Carry out lyophilization in the microsphere suspension immigration freeze dryer of step d) with step C gained, make dried frozen aquatic products.
Before step e) is faced usefulness, add sterile water for injection, dried frozen aquatic products is redissolved, promptly get the microsphere aqueous suspension.
13, according to the preparation contrast agent method of claim 12, wherein said processor for ultrasonic wave is that frequency is the probe type ultrasonoscope of 10～50KHz.
14, according to the preparation contrast agent method of claim 12, wherein said shearing equipment is homogenizer or colloid mill.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710106559 CN101062423A (en) | 2007-06-06 | 2007-06-06 | Gas emulsion type Ultrasound Contrast Agent microspheres and the preparing method thereof |
| CN2008100986918A CN101596322B (en) | 2007-06-06 | 2008-06-06 | Aerial emulsion type ultraphonic contrast agent microballoon and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710106559 CN101062423A (en) | 2007-06-06 | 2007-06-06 | Gas emulsion type Ultrasound Contrast Agent microspheres and the preparing method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101062423A true CN101062423A (en) | 2007-10-31 |
Family
ID=38963838
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200710106559 Pending CN101062423A (en) | 2007-06-06 | 2007-06-06 | Gas emulsion type Ultrasound Contrast Agent microspheres and the preparing method thereof |
| CN2008100986918A Expired - Fee Related CN101596322B (en) | 2007-06-06 | 2008-06-06 | Aerial emulsion type ultraphonic contrast agent microballoon and preparation method thereof |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100986918A Expired - Fee Related CN101596322B (en) | 2007-06-06 | 2008-06-06 | Aerial emulsion type ultraphonic contrast agent microballoon and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN101062423A (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105536000B (en) * | 2015-12-10 | 2018-11-27 | 北京大学 | A kind of acoustic contrast agent and its preparation method and application based on pentaerythritol ester |
| CN106943606A (en) * | 2017-03-15 | 2017-07-14 | 贵州灿石正科信息咨询服务有限公司 | A kind of acoustic contrast agent |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5656211A (en) * | 1989-12-22 | 1997-08-12 | Imarx Pharmaceutical Corp. | Apparatus and method for making gas-filled vesicles of optimal size |
| GB9106673D0 (en) * | 1991-03-28 | 1991-05-15 | Hafslund Nycomed As | Improvements in or relating to contrast agents |
| US6572840B1 (en) * | 1999-07-28 | 2003-06-03 | Bristol-Myers Squibb Pharma Company | Stable microbubbles comprised of a perfluoropropane encapsulated lipid moiety for use as an ultrasound contrast agent |
| CN1194763C (en) * | 2002-09-06 | 2005-03-30 | 中国人民解放军第三军医大学 | Novel lipide supersonic contrast medium and preparation method thereof |
| CN1321697C (en) * | 2003-12-23 | 2007-06-20 | 中国人民解放军军事医学科学院毒物药物研究所 | Ultrasound contrast medium composition with phospholipid as membrane material and its preparation method |
-
2007
- 2007-06-06 CN CN 200710106559 patent/CN101062423A/en active Pending
-
2008
- 2008-06-06 CN CN2008100986918A patent/CN101596322B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN101596322A (en) | 2009-12-09 |
| CN101596322B (en) | 2011-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1268397C (en) | Method for enhancing echogenicity and decreasing attenuation of microencapsulated gases | |
| CN101227890B (en) | Method and system for in vivo drug delivery | |
| EP0504340B1 (en) | Method for the preparation of stable suspensions of hollow gas-filled microspheres suitable for ultrasonic echography | |
| CN1055413C (en) | Stable microbubbles suspensions injectable into living organisms | |
| EP0907380B1 (en) | Pressure resistant protein microspheres as ultrasonic imaging agents | |
| US20120034170A1 (en) | Nanobubbles Useful As An Ultrasonic Contrast Agent For The Lymphatic System | |
| KR101595795B1 (en) | Dual-Purpose PAT/Ultrasound Contrast Agent with Nanoparticles Including Drug and Method for Preparing the Same | |
| JPH08502979A (en) | Surfactant-stabilized microbubble mixture, process for its production and use thereof | |
| JP2008509970A (en) | Gas-filled microvesicle composition for contrast imaging | |
| JP4829796B2 (en) | Ultrasound contrast agent | |
| CN101780284B (en) | Magnetic molecular targeted ultrasound contrast agent microsphere and preparation method thereof | |
| US6572840B1 (en) | Stable microbubbles comprised of a perfluoropropane encapsulated lipid moiety for use as an ultrasound contrast agent | |
| HUT67156A (en) | Encapsulated particles useful as contrast agent in ultrasound and x-ray imagong and process for production of them | |
| CN107625970A (en) | A kind of acoustic radiation force strengthens the bionical microvesicle complex enrichment of inflammatory cell atherogenicity and targeted system | |
| CN101596322B (en) | Aerial emulsion type ultraphonic contrast agent microballoon and preparation method thereof | |
| JPH07507778A (en) | Use of microcapsules as contrast agents for color Doppler sonography | |
| CN105536000B (en) | A kind of acoustic contrast agent and its preparation method and application based on pentaerythritol ester | |
| CN101637613B (en) | Lipid targeted ultrasound contrast agent and preparation method thereof | |
| CN1993147A (en) | Ultrasonic contrast agent dosage formulation | |
| EP1875928A1 (en) | Novel method of using triacetin and auxiliary agent for ultrasonic diagnostic examination | |
| JPH08310971A (en) | Contrasting agent for ultrasonic diagnosis | |
| KR20230126419A (en) | Ultrasound contrast agent including micronano bubbles and manufacturing method thereof | |
| CN101732735A (en) | Gas emulsion type ultrasound contrast agent microspheres using carbohydrate as main material and preparation method thereof | |
| US20030206862A1 (en) | Contrast medium based on polyoxyethylene-660-12-hydroxystearate and anionic phospholipids | |
| Smith | In vitro Characterization of Echogenic Liposomes (ELIP) for Ultrasonic Delivery of Recombinant Tissue-type Plasminogen Activator (rt-PA) |
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 |