WO1997026016A2 - Suspensions de bulles de gaz et leur utilisation sous forme d'agents de contraste ultrasonores - Google Patents

Suspensions de bulles de gaz et leur utilisation sous forme d'agents de contraste ultrasonores Download PDF

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Publication number
WO1997026016A2
WO1997026016A2 PCT/EP1997/000208 EP9700208W WO9726016A2 WO 1997026016 A2 WO1997026016 A2 WO 1997026016A2 EP 9700208 W EP9700208 W EP 9700208W WO 9726016 A2 WO9726016 A2 WO 9726016A2
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WIPO (PCT)
Prior art keywords
water
fatty acid
porous matrix
mol
acid esters
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PCT/EP1997/000208
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German (de)
English (en)
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WO1997026016A3 (fr
Inventor
Martina Bergmann
Dieter Heldmann
Werner Weitschies
Thomas Fritzsch
Violetta Sudmann
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Schering Aktiengesellschaft
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Priority to AU15925/97A priority Critical patent/AU1592597A/en
Priority to JP9525697A priority patent/JP2000504317A/ja
Priority to EP97902178A priority patent/EP0874644A2/fr
Publication of WO1997026016A2 publication Critical patent/WO1997026016A2/fr
Publication of WO1997026016A3 publication Critical patent/WO1997026016A3/fr
Priority to NO983332A priority patent/NO983332L/no

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/04—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent
    • C08J9/12—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent
    • C08J9/14—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent organic
    • C08J9/143—Halogen containing compounds
    • C08J9/147—Halogen containing compounds containing carbon and halogen atoms only
    • C08J9/148—Halogen containing compounds containing carbon and halogen atoms only perfluorinated
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
    • A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
    • A61K49/223—Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/04—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent
    • C08J9/12—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent
    • C08J9/14—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent organic
    • C08J9/143—Halogen containing compounds
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2300/00—Characterised by the use of unspecified polymers

Definitions

  • the invention relates to the subject characterized in the claims, that is to say porous matrices for generating stable gas bubble suspensions, their use as ultrasound contrast agents and methods for producing the matrices and agents.
  • Ultrasound diagnostics offer the possibility of diagnosing physiological and pathophysiological conditions without stressful ionizing radiation as in X-ray or radionuclide examinations and relatively inexpensively compared to magnetic resonance imaging.
  • Ultrasonic waves are reflected or absorbed depending on the acoustic properties of the tissue. Different acoustic properties of tissues and body fluids are used for imaging. Due to the large density difference between body tissue or body fluids on the one hand and gas bubbles on the other hand, gases in the form of microbubbles are particularly suitable as contrast agents for ultrasound. Ultrasound contrast agents are therefore essentially researched and developed on the basis of gas bubbles and / or substances containing gas.
  • the simplest type of ultrasound contrast medium can be obtained by methods such as shaking, sonicating or pumping around an aqueous suspension medium between two syringes.
  • the introduced bubbles can be made by suitable
  • Additives such as surfactants and / or viscosity-increasing substances are stabilized.
  • Such contrast agents are e.g. described in EP 0 077 752.
  • the strong dependence of the number and size of gas bubbles on the type, duration and intensity of the agitation is problematic. This makes the production difficult to reproduce and the risk of an embolism due to excessive gas bubbles is uncontrollable.
  • a similar type of contrast medium is described in WO 93/05819, but instead of the atmospheric gases such as air, nitrogen, carbon dioxide and noble gases, gases with a certain Q factor are proposed for the preparation of the bladder suspensions.
  • gases with a certain Q factor are proposed for the preparation of the bladder suspensions.
  • gases usually halogenated hydrocarbons that are characterized by low solubility in physiological media.
  • Perfluorinated compounds in particular are suitable as exchange gases.
  • these agents show an inhomogeneous and poorly reproducible bubble size distribution. This increases the risk of embolism due to gas bubbles that are too large.
  • solid carriers can also be formulated, from which bubbles are released after resuspending in a suitable diluent.
  • a microparticle suspension is used here.
  • Such solid carriers can be microparticles which consist, for example, of a mixture of at least one surface-active substance with at least one non-surface-active solid, as are disclosed in EP 0 365 467.
  • non-surface-active solids can also be substances which are used as X-ray contrast media (WO 93/00930 and WO 92/21382), in the case of the latter document the X-ray contrast media being crosslinked via functional groups and crosslinkers .
  • microparticle suspensions mentioned are ultrasound contrast media with which contrast effects can be achieved in the arterial system. Contrast intensity and duration seem to be in need of improvement.
  • microparticulate ultrasound contrast agents are disclosed in WO 95/21631.
  • water-insoluble wall formers are dissolved in an organic solvent (toluene), then emulsified in an aqueous surfactant solution and freeze-dried.
  • an organic solvent toluene
  • emulsified in an aqueous surfactant solution By resuspending a microparticle suspension is obtained which shows ultrasound contrast in vivo.
  • a microparticle suspension which shows ultrasound contrast in vivo.
  • the use of certain fluorinated substances is also described for the type of microparticulate ultrasound contrast media.
  • EP 0 554 213 discloses galactose-based microparticles which contain SFg instead of air.
  • the contrast-enhancing effects for this remedy are weak.
  • WO 95/22994 also discloses microparticles containing fluorinated substances.
  • the bubble size distribution is standardized and the number of bubbles significantly increased, see above that the dose required for an ultrasound contrast agent application can be considerably reduced.
  • WO 95/03835 claims ultrasound contrast agents based on particles which contain defined gas mixtures.
  • the gas mixtures consist of at least one fluorinated, gas-osmotically active component and at least one conventional gas such as nitrogen, oxygen and / or carbon dioxide.
  • the particles are made up of proteins, dextrins, starch and starch derivatives, e.g. Hydroxyethyl starch.
  • those with a high molecular weight > 500,000 daltons
  • those with a high molecular weight > 500,000 daltons
  • these substances cannot be filtered through the kidneys and must be broken down by the liver, among other things. This increases the dwell time in the body.
  • hydroxyethyl starch is broken down into substituted oligosaccharides which are primarily eliminated renally when the kidney threshold is undershot.
  • the storage of hydroxyethyl starch in the cells of the reticuloendothelial system is discussed as a possible problem.
  • the ethyl ether bond is not amenable to enzymatic degradation.
  • the metabolism and elimination of hydroxyethylglucose, as well as their possible pharmacological effects, is currently not clear [Krech, I .; Wind, S. (1995) Wettzie 16 (2), 62-63].
  • higher molecular weight substances carry the risk that particulate portions of uncontrolled size are applied after resuspending the contrast medium preparation. The patient is at risk of embolism.
  • Fluorinated gases and air-containing particles are also claimed in WO 95/16467, the proportion of the fluorinated component here being limited to 41%.
  • WO 94/09829 describes ultrasound contrast agents containing liposomes.
  • Phospholipids but also other surfactants which are difficult to dissolve in water are mentioned as gas bubbles stabilizing surfactant.
  • Such liposomal systems are generally produced by lyophilization from freeze-dry solvents such as, for example, tertiary-butanol or C 2 Cl 4 F 2 .
  • the use of organic solvents to produce these agents requires a lot of effort
  • Chlorinated hydrocarbons such as C 2 C1 4 F 2 are also extremely critical because of their ozone depleting potential.
  • the object of the present invention was therefore to provide compounds for the production of ultrasound contrast media which overcome the disadvantages of the prior art, i.e. the
  • preparations consisting of a porous, solid, water-soluble matrix containing a low molecular weight scaffold, a surfactant and a gas, the gas being enclosed in the pores of the matrix, are outstandingly suitable for producing a preparation for ultrasound diagnosis.
  • the contrast media according to the invention are generated from a particle-free, porous, solid structure, which is referred to below as a porous matrix.
  • FIGS. 1 and 2 show an image of a preparation according to the invention produced according to Example 19 at the same magnification (1 cm in the figures corresponds to 1.1 ⁇ m in reality) .
  • Clearly recognizable are the uniform pores from which gas bubbles are released after the matrix is dissolved. The size and number of pores are highly reproducible. The size of the bubbles is essentially limited by the pore size.
  • the number of bubbles that can be released from the matrix is also determined by the porosity of the matrix.
  • the parameters mentioned can be easily controlled via various production parameters and have an important influence on the effectiveness of the contrast medium.
  • the formation of the gas bubbles is not linked to agitation of the medium before application, so that the gas bubbles can be released in unchanged form.
  • the gas bubbles released are particularly advantageous with the help of surfactants. which may be part of the matrix, can be stabilized.
  • a stabilized gas bubble suspension is generally used.
  • the porous matrices according to the invention are made up of a water-soluble scaffold, which generally has a molecular weight ⁇ 15,000 Daltons, and a rapidly and readily water-soluble surfactant, the surfactant content in the matrix being 0.01 to 10% (m / m) .
  • a water-soluble scaffold which generally has a molecular weight ⁇ 15,000 Daltons, and a rapidly and readily water-soluble surfactant, the surfactant content in the matrix being 0.01 to 10% (m / m) .
  • Examples include L-glycine, L-alanine, L-valine, L-leucine, L-isoleucine, L-phenylalanine, L-proline, L-hydroxyproline, L-serine, L-threonine, L-tryptophan, L-asparagine , L-glutamine, L-arginine, L-histidine, glycyl-glycine, glycyl-glycyl-glycine, glucose, galactose, fructose, mannose, sorbose, sucrose, lactose, maltose, trehalose, gentiobiose, lactulose, turanose, maltotriose, melibiose
  • X-ray contrast media and contrast media for magnetic resonance imaging are also suitable as scaffold builders.
  • examples include iopromide, iotrolan, iopamidol, iohexol, and Gd-DTPA (gadopentetic acid), Gd-DTPA-dimeglumine salt (Magnevist), Gd-EOB-DTPA (gadoxetic acid, disodium salt) and gadobutrol.
  • Suitable surfactants are water-soluble, nonionic surfactants, those with a perfluorinated hydrocarbon building block and / or with a molecular weight of ⁇ 15,000 daltons being preferred.
  • Examples include sorbitan fatty acid esters, polyoxyethylene sorbitan, polyoxyethylene sorbitol, polyoxyethylene fatty acid esters, Glycerinpolyoxyethylenfettklar, ethoxylated mono-, di-, triglycerides, which if desired, may partially be hydrogenated and ethoxylated mixtures thereof, ethoxylated castor oils, ethoxylated phenols, polyoxyethylene fatty alcohol ethers, polyglycerol fatty acid ester, Sorbitanperfluorfettkladreester, Polyoxyethylensorbitanperfluorfettklar, Polyoxyethylensorbitolperfluorfettklaester, Polyoxyethylene perfluoro fatty acid esters, poly(
  • phospholipids especially hydrogenated phosphatipy Icho lin.
  • the gases used are in particular fluorinated gases.
  • fluorinated gases Surprisingly, stronger and longer lasting in vivo contrast effects are observed when using the "classic" gases than are achieved with the particulate preparations of the prior art.
  • fluorinated gases it is surprisingly possible to dispense with the use of gas mixtures, as are required in the preparations of the prior art.
  • Tetrafluoroallenes hexafluoro-1,3-butadiene, decafluorobutane, perfluoro-1-butenes, perfluoro-2-butenes, perfluoro-2-butyne, octafluorocyclobutane, perfluorocyclobutene, perfluorocyclopentane, perfluorodimethylamine, hexafluoroethane, tetrafluoromethane, tetrafluoromethane, tetrafluoromethane, tetrafluoromethylene Perfluoropropane and perfluoropropylene.
  • the perfluorinated substances are particularly preferred.
  • the matrices according to the invention can be produced with less effort under aseptic conditions by first providing an aqueous scaffold solution, to which gas bubble-stabilizing surfactants are added with particular advantage.
  • the separated or combined solutions can first be sterile filtered, then the mixture thus prepared is quickly frozen.
  • the removal of the water takes place under conditions which allow a direct transition of the ice into the gaseous state without passing through the liquid state of matter. Possible suitable conditions can be found in the phase diagram of the water (see FIG. 3). What remains is a porous matrix that is aerated with the desired gas.
  • For complete gas exchange i.e.
  • a particular advantage of the process according to the invention is that the use of organic solvents can be dispensed with. There is also no need for technologically complex processing, as would be necessary in the case of poorly or slowly soluble or only water-dispersible substances. Residual solvent contents as a critical quality feature therefore play a role for the agents according to the invention do not matter. This is a significant advantage both from an ecological perspective and in terms of product safety. Many organic solvents, even in the smallest amounts or concentrations, are suspected of being carcinogenic and / or mutagenic.
  • the desired particle-free ultrasound contrast agents can easily be produced from the matrices according to the invention by adding an aqueous medium.
  • the treating physician adds the aqueous medium immediately before use.
  • the aqueous medium can contain the auxiliaries customary in galenics, e.g. Isotonizing and viscosity-increasing additives. It is not necessary to shake the fluidized matrix.
  • the contrast media thus produced are distinguished by the fact that they are blood-isotonic or almost blood-isotonic. They can be injected immediately after resuspending.
  • the concentration of the contrast media is 10 to 600 mg, preferably 50 to 400 mg of matrix material per milliliter of suspension.
  • the agents are administered in a dose of 0.01 to 0.20 ml / kg of body weight.
  • the agents according to the invention are for all imaging modes of sonography, such as. B. M, B, Doppler mode but also for modes in which nonlinear effects are used such as Harmony and harmony power mode equally suitable and can be produced with high reproducibility.
  • the bubble numbers generated from the matrix are significantly higher than those of the prior art agents, the agents therefore show significantly improved contrast effects; in particular, the time window available for the examination could be significantly extended (see also in-vivo experiments 41-52).
  • Example 5 30 g of raffinose (MW: 594 g / mol) are mixed with 0.3 g Zonyl ® FSO-100 (MW: - 725 g / mol) and 70 g water. The procedure is then as described in Example 1. After resuspending in 10 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 262 mosmol. 14.6xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 5 30 g of raffinose (MW: 594 g / mol) are mixed with 0.3 g Zonyl ® FSO-100 (MW: - 725 g / mol) and 70 g water. The procedure is then as described in Example 1. After resuspending in 10 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 262 mosmol. 14.6xl
  • Gd-EOB-DTPA (MW: 726 g / mol) (Gadoxetic acid, disodium) are mixed with 0.3 g Zonyl ® FSO-100: - were added and 70 g of water (MW 725 g / mol). The mixture is stirred until completely dissolved. The solution is filled with 3 g and frozen with liquid nitrogen. After the water has been completely removed, one remains porous matrix. When resuspended in 10 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 344 mosmol. 24.6xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Gadobutrol (MW: 605 g / mol) are mixed with 0.3 g Zonyl ® FSO-100 (MG: - 725 g / mol) and 70 g water. The mixture is stirred until completely dissolved. The solution is filled with 3 g and frozen with liquid nitrogen. After the water has been completely removed, a porous matrix remains. When resuspended in 5 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 269 mosmol. 20, lxl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • gadopentetic acid (MW: 548 g / mol) are mixed with 0.3 g Zonyl ® FSO-100 (MW: - 725 g / mol) and 70 g water. The mixture is stirred until completely dissolved. The solution is filled with 3 g and frozen with liquid nitrogen. After the water has been completely removed, a porous matrix remains. When resuspended in 7 ml of water, a preparation is obtained which isotonic with an osmolality of 282 mosmol. 29, lxl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 20 40 g iopromide (MW: 791 g / mol) are mixed with 0.4 g Zonyl ® FSO-100 (MW: - 725 g / mol) and 60 g water. The procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 289 mosmol. 22.4xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 20
  • Osmolality of 342 mosmol is almost isotonic. 6, 16 ⁇ 10 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • iopromide MW: 791 g / mol
  • Triton® -X-100 MW: - 874 g / mol
  • the procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 290 mosmol. 9.56xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • iopromide MW: 791 g / mol
  • Rewoderm ® Li 48-50 MW: - 3800 g / mol
  • the procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 288 mosmol. 24.05xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • iopromide MW: 791 g / mol
  • Solutol ® HS 15 MW: - 1000 g / mol
  • Example 1 After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 289 mosmol. 13.46xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 28 40 g iopromide (MW: 791 g / mol) are mixed with 0.4 g of Lutrol ® F68 (MW - 8600 g / mol) and 60 g water. The procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 290 mosmol. 17.68xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 28 40 g iopromide (MW: 791 g / mol) are mixed with 0.4 g of Lutrol ® F68 (MW - 8600 g / mol) and 60 g water. The procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation isotonic with an osmolality of 290 mosmol. 17.68xl0 9 bubbles in the range of 0.56-7.46
  • iopromide MW: 791 g / mol
  • Span ® 85 MW: 1028 g / mol
  • the procedure is then as described in Example 1. After resuspending in 6 ml of water, a preparation is obtained which is isotonic with an osmolality of 291 mosmol. 20.45xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Gadobutrol (MW: 605 g / mol) are mixed with 0.3 g Solutol ® HS 15 (MG: - 1000 g / mol) and 70 g water. The mixture is stirred until completely dissolved. The solution is filled with 3 g and frozen with liquid nitrogen. After the water has been completely removed, a porous matrix remains. When resuspended in 5 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 271 mosmol. 6.64xl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • iopromide MW: 791 g / mol
  • Triton® -X-100 MW: 874 g / mol
  • the procedure is then as described in Example 1.
  • a gas exchange with hexafluoroethane is carried out.
  • a preparation is obtained which is isotonic with an osmolality of 290 mosmol. 3.01 ⁇ 10 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Gadobutrol (MW: 605 g / mol) are mixed with 0.3 g Solutol ® HS 15 (MG: - 1000 g / mol) and 70 g water. The mixture is stirred until completely dissolved. The solution is filled with 3 g and frozen with liquid nitrogen. The procedure is then as described in Example 1. After drying, a gas exchange with hexafluoroethane is carried out. When resuspended in 5 ml of water, a preparation is obtained which is almost isotonic with an osmolality of 271 mosmol. 2, llxl0 9 bubbles in the range of 0.56-7.46 ⁇ m are released per gram of substance.
  • a beagle dog [female, 11.2 kg body weight (hereinafter KGW)] is anesthetized (inhalation anesthesia approx. 2/3 oxygen, approx. 1/3 N 2 O 1.5 - 2% enflurane; spontaneous breathing) and for a sonographic examination of the heart prepared.
  • the examination is carried out using an HP ultrasound system (type 77020 E, 5 MHz transducer) in B mode.
  • the test animal receives an intravenous application of the test substance (agent according to the invention produced according to Example 22).
  • a contrast medium which was produced analogously to WO 95/22994 (Example 12), serves as the reference substance.
  • the doses used are 0.1 ml / kg body weight both for the agent according to the invention and for the reference substance.
  • the result is shown in the form of the intensity-time profiles in FIG. 4.
  • the upper (slower falling) curve corresponds to the preparation according to the invention - as also in the following FIGS. 5-8. It can be clearly seen that the agents according to the invention show a longer lasting contrast after intravenous injection than the agents of the prior art. These contrasting properties give the doctor longer periods of time for the examination (examination window). Furthermore, the
  • Example 12 Preparation prepared as described in Example 27), an agent from WO 95/22994 (Example 12) was used as reference.
  • the doses for reference and test substance were identical and each was 0.1 ml / kg body weight.
  • the intensity-time profiles are shown in FIG. 5.
  • Example 44 The procedure is as described in Example 41, but the test animal had a body weight of 11.9 kg.
  • a preparation prepared as described in Example 28) serves as the test substance, and an agent from EP 0365467 (Example 1) was used.
  • the doses used were 0.05 ml / kg body weight for the agent according to the invention according to Example 28 and 0.2 ml / kg body weight for the reference.
  • the result of the examination is shown in the form of the intensity-time profiles in FIG. 6. In this case too, despite the low dose, a more intense and longer-lasting contrast is observed for the preparation according to the invention.
  • Example 44 Example 44
  • the doses used were 0.05 ml / kg body weight for the agent according to the invention and 0.2 ml / kg body weight for the reference.
  • the result of the examination is shown in the form of the intensity-time profiles in FIG. 7.
  • a beagle dog female, 9.7 kg body weight
  • a preparation prepared according to Example 19 serves as the test substance, and an agent from WO 95/22994 (Example 12) was used as a reference.
  • the doses for reference and test substance were 0.1 ml / kg body weight.
  • the intensity-time profiles are shown in FIG. 8.
  • Example 12 Preparation prepared as described in Example 5), an agent from WO 95/22994 (Example 12) was used as reference.
  • the doses for reference and test substance were identical and were 0.1 ml / kg body weight.
  • FIG. 9 The result of the investigation is shown in FIG. 9. The heart of the dog was shown. In detail show:
  • Example 41 The procedure is as described in Example 41.
  • a preparation prepared as described in Example 11) serves as the test substance, and an agent from WO 95/22994 (Example 12) was used as reference.
  • the doses for reference and test substance were identical and were also 0.1 ml / kg body weight.
  • a beagle dog female, 9.6 kg body weight
  • is anesthetized inhalation anesthesia approx. 2/3 oxygen, approx. 1/3 N 2 O, 1.5 - 2% enflurane, spontaneous breathing
  • the examination is carried out using an HP ultrasound system (type 77020 E, 5 MHz transducer) in B mode.
  • the test animal each receives an intravenous application of an agent according to the invention produced according to one of Examples 31, 35, 39 and, as a reference, an injection of a contrast agent according to the prior art, which was produced analogously to WO 95/11994 (Example 12).
  • the doses used were 0.1 ml / kg body weight for the agents according to the invention and for the reference. The result is shown in the form of the intensity values in Table 1. It is also clearly evident here that the agents according to the invention are a show significantly higher contrast levels after intravenous injection than the reference agent.
  • a beagle dog female. 9.7 kg body weight
  • is anesthetized inhalation anesthesia approx. 2/3 oxygen, approx. 1/3 N 2 O, 1.5 - 2% enflurane, spontaneous breathing
  • the examination is carried out using an HP ultrasound system (type 77020 E, 5 MHz transducer) in B mode.
  • the test animal each receives an intravenous application of an agent according to the invention produced according to one of Examples 4, 8, 9 and, as a reference, an injection of a contrast agent according to the prior art, which was produced analogously to WO 95/22994 (Example 12).
  • the doses used were 0.1 ml / kg body weight for the agents according to the invention and for the reference.
  • the result is shown in Table 2 in the form of the area under the intensity-time curve (density units x see). It is clearly recognizable that the Agents according to the invention show a higher area level after intravenous injection.
  • a beagle dog male, 15.5 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen, 1-3% enflurane, rest nitrogen; spontaneous breathing
  • the examination takes place with the ultrasonic system ATL UM-9 with the Tranducer type L 10-5.
  • the test animal each receives an intravenous application of the test substance produced according to Example 8 or 19 and, as a reference, an application of a contrast agent produced according to WO 95/22994 (Example 12).
  • the dose for all injections was 0.1 ml / ks body weight.
  • the spectral duplication signal is evaluated intensitometrically and plotted against time. The resulting areas under the intensity-time curves are shown in Table 3.
  • a beagle dog female, 9.7 kg body weight
  • is anesthetized inhalation anesthesia approx. 2/3 O 2 : approx. 1/3 N 2 O; 1.5 - 3% enflurane, spontaneous breathing
  • the examination is carried out using an HP ultrasound system (type Sonos 1000, 5MHz) in color Doppler mode.
  • the experimental animal receives an intravenous application of an agent according to the invention according to Example 22 (0.1 ml / kg body weight). After administration, the perfusion display of the organ is significantly improved compared to the display before application.
  • a beagle dog female, 9.7 kg body weight
  • is anesthetized inhalation anesthesia approx. 2/3 O 2 ; approx. 1/3 N 2 O; 1, 5 - 3% enflurane, spontaneous breathing
  • sonographic examination of the abdominal aorta prepared.
  • the examination is carried out with an ultrasound system of the brand ATL type UM9 with transducer C10-5 in harmony B- Fashion.
  • the experimental animal receives an intravenous application of an agent according to the invention prepared according to Example 8 (dose 0.1 ml / kg body weight).
  • the vascular volume is marked echogenically. Before the injection, the volume was without enhancement.
  • sucrose palmitate stearate 7 0.67 g of sucrose palmitate stearate 7 are mixed with 100 g of water and heated in the micro wave. After cooling, a slightly cloudy, stable solution is obtained. 60 g of this opalescent solution are mixed with 40 g iopromide (MW: 791 g / mol). The procedure is then as described in Example 1. After resuspending in 6 ml of water, an isotonic preparation is obtained. 62.01 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • iopromide MW: 791 g / mol
  • sucrose palmitate stearate 7 0.67 g of sucrose palmitate stearate 7 are mixed with 100 g of water and heated in the microwave. After cooling, a slightly cloudy, stable solution is obtained. 60 g of this opalescent solution are mixed with 40 g iopromide (MG.-791 g / mol). The procedure is then as described in Example 1. The gas exchange takes place with perfluorohexane. After resuspending in 6 ml of water, an isotonic preparation is obtained. 2.82 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • sucrose palmitate stearate 7 0.67 g of sucrose palmitate stearate 7 are mixed with 100 g of water and heated in the microwave. After cooling, a slightly cloudy, stable solution is obtained. 60 g of this opalescent solution are mixed with 40 g iopromide (MW: 791 g / mol). The mixture is stirred until completely dissolved. The entire solution is frozen by dropping it in liquid decafluorobutane. After removal of the water under conditions which allow a direct transition from the solid to the gaseous state without the liquid state being passed through, the product is filled with 1 g and a gas exchange is carried out with decafluorobutane. After resuspending in 3 ml of water, an isotonic preparation is obtained. 34.07 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 59 0.4 g of hydrogenated phosphatipylcholine (Pro Lipo H ® ) are mixed with 60 g of water and then with 0.4 g of iopromide (MW: 791 g / mol). The procedure below is as described in Example 1. After resuspending in 6 ml of water, an isotonic preparation is obtained. 27.97 x 10 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 59 0.4 g of hydrogenated phosphatipylcholine (Pro Lipo H ® ) are mixed with 60 g of water and then with 0.4 g of iopromide (MW: 791 g / mol). The procedure below is as described in Example 1. After resuspending in 6 ml of water, an isotonic preparation is obtained. 27.97 x 10 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • sucrose palmitate stearate 7 1.5 g of sucrose palmitate stearate 7 are mixed with 100 g of water and heated in the microwave. After cooling, a slightly cloudy, stable solution is obtained. 70 g of this opalescent solution are mixed with 30 g maltooligosaccharide (MW: 684 g / mol) and 0.1 g PVA (MG: 10000 g / mol). The preparation is filled at 4 g. The procedure is then as described in Example 1. After resuspending in 6 ml of water, an isotonic preparation is obtained. 9.37 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • 40 g iopromide (MW: 791 g / mol) are mixed with 0.04 g of polyoxyethylene sorbitan monolaurate Tween 21 ®) and 60 g of water. The mixture is stirred until completely dissolved. The preparation is filled with 5 g and frozen in liquid propane. After removal of the water under conditions that allow a direct transition from the solid to the gaseous state without the liquid If the physical state is exceeded, a gas exchange with decafluorobutane is carried out. After resuspending in 6 ml of water, an isotonic preparation is obtained. 3.77 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • sucrose palmitate stearate 7 0.67 g of sucrose palmitate stearate 7 are mixed with 100 g of water and heated in the microwave. After cooling, a slightly cloudy, stable solution is obtained. 60 g of this opalescent solution are mixed with 40 g iopromide (MW: 791 g / mol). The preparation is filled with 5 g and frozen in liquid butane. After removal of the water under conditions which enable a direct transition from the solid to the gaseous state without passing through the liquid state of matter, a gas exchange with decafluorobutane is carried out. After resuspending in 6 ml of water, an isotonic preparation is obtained. 34.16 x IO 9 bubbles in the contrast-relevant range of 0.56-7.46 ⁇ m are released per gram of substance.
  • Example 66 The procedure is then as described in Example 1. After resuspending in 2.5 ml of water, an isotonic preparation is obtained.
  • Example 66
  • a beagle dog female, 11.8 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen. 1 - 3% enflurane. Rest nitrogen;
  • a beagle dog female, 11.9 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen, 1 - 3% enflurane, rest nitrogen; spontaneous breathing
  • the examination is carried out with the ultrasound system ATL UM-9 with the transducer type L 10-5.
  • the test animal each receives an intravenous application of the test substance produced according to Example 58 (0.05 ml / kg) and, as a reference, an application of a contrast agent produced according to EP 0365467 (Example 1, 0.2 ml / kg).
  • Figure 12 shows the intensity-time profiles of the two injections. The clearly more intense and longer lasting contrast of the preparation according to the invention is clearly recognizable.
  • a beagle dog female, 12.1 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen, 1 - 3% enflurane, remainder nitrogen; spontaneous breathing
  • the examination is carried out with the ultrasound system ATL UM-9 with the transducer type L 10-5.
  • the test animal each receives an intravenous application of the test substance produced according to Example 59 (0.05 ml / kg) and, as a reference, an application of a contrast agent produced according to EP 0365467 (example 1, 0.2 ml / kg).
  • the area under the intensity-time curve is 252.2% of the reference.
  • a beagle dog female, 12.1 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen, 1 - 3% enflurane, remainder nitrogen; spontaneous breathing
  • the examination is carried out with the ultrasound system ATL UM-9 with the transducer type L 10-5.
  • the test animal each receives an intravenous application of the test substance produced according to Example 64 (0.05 ml / kg) and, as a reference, an application of a contrast agent produced according to EP 0365467 (Example 1, 0.2 ml / kg).
  • the area under the intensity-time curve is 223.8% of the reference.
  • a beagle dog male, 17.1 kg body weight
  • is anesthetized inhalation anesthesia 23% oxygen, 1 - 3% enflurane, rest nitrogen; spontaneous breathing
  • the examination is carried out with the ultrasound system ATL UM-9 with the transducer type L 10-5.
  • the test animal each receives an intravenous application of the test substance produced according to Example 65 (0.05 ml / kg) and, as a reference, an application of a contrast agent produced according to EP 0 365 467 (Example 1, 0.2 ml / kg).
  • the area under the intensity-time curve is 181.0% of the reference.

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Abstract

L'invention concerne des matrices poreuses constituées de substances de faible poids moléculaire servant à générer des suspensions de bulles de gaz stables, leur utilisation sous forme d'agents de contraste ultrasonores et des procédés de production de ces matrices et agents de contraste.
PCT/EP1997/000208 1996-01-18 1997-01-16 Suspensions de bulles de gaz et leur utilisation sous forme d'agents de contraste ultrasonores WO1997026016A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AU15925/97A AU1592597A (en) 1996-01-18 1997-01-16 Gas bubble suspensions and their application as an ultrasound contrast medium
JP9525697A JP2000504317A (ja) 1996-01-18 1997-01-16 気泡懸濁物と超音波造影剤への応用
EP97902178A EP0874644A2 (fr) 1996-01-18 1997-01-16 Suspensions de bulles de gaz et leur utilisation sous forme d'agents de contraste ultrasonores
NO983332A NO983332L (no) 1996-01-18 1998-07-17 Gassboblesuspensjoner og deres anvendelse som ultralydkontrastmiddel

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19602930.9 1996-01-18
DE19602930A DE19602930A1 (de) 1996-01-18 1996-01-18 Poröse Matrices aus niedermolekularen Substanzen zur Genierung stabiler Gasblasensuspensionen, deren Verwendung als Ultraschallkontrastmittel sowie Verfahren zu deren Herstellung

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WO1997026016A2 true WO1997026016A2 (fr) 1997-07-24
WO1997026016A3 WO1997026016A3 (fr) 1997-10-23

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AU (1) AU1592597A (fr)
CA (1) CA2243174A1 (fr)
DE (1) DE19602930A1 (fr)
NO (1) NO983332L (fr)
WO (1) WO1997026016A2 (fr)
ZA (1) ZA97414B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20240226418A1 (en) * 2021-01-26 2024-07-11 Agitated Solutions Inc. Syringe-Based Microbubble Generator

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NZ592293A (en) * 2005-11-28 2012-10-26 Verrow Pharmaceuticals Inc Compositions comprising an oligosaccharide such as a cyclodextrin for reducing the nephrotoxicity of other drugs
JP5196896B2 (ja) * 2007-07-13 2013-05-15 花王株式会社 微細気泡前駆体の製造方法
JP2010005512A (ja) * 2008-06-25 2010-01-14 Kao Corp 微細気泡前駆体組成物
KR101927614B1 (ko) 2017-03-30 2018-12-10 숭실대학교산학협력단 초음파 영상을 위한 안정한 생체적합 산소-나노버블 조영제 제조방법
JP7211699B2 (ja) * 2017-09-11 2023-01-24 バイエル ファーマ アクチエンゲゼルシャフト 超音波診断用造影剤

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EP0123235A2 (fr) * 1983-04-15 1984-10-31 Schering Aktiengesellschaft Microparticules et milieu de contraste pour l'exploration ultrasonique contenant des bulles de gaz
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EP0122624A2 (fr) * 1983-04-15 1984-10-24 Schering Aktiengesellschaft Microparticules et milieu de contraste pour l'exploration ultrasonique contenant des bulles de gaz
EP0123235A2 (fr) * 1983-04-15 1984-10-31 Schering Aktiengesellschaft Microparticules et milieu de contraste pour l'exploration ultrasonique contenant des bulles de gaz
US5160725A (en) * 1987-03-24 1992-11-03 Silica Gel Gesellschaft Mbh Adsorptions-Technik, Apparatebau Magnetic liquid compositions
EP0365467A2 (fr) * 1988-10-07 1990-04-25 Schering Aktiengesellschaft Agent de contraste ultrasonique à base de bulles de gaz et de microparticules contenant des acides gras
US5147631A (en) * 1991-04-30 1992-09-15 Du Pont Merck Pharmaceutical Company Porous inorganic ultrasound contrast agents
WO1992021382A1 (fr) * 1991-06-03 1992-12-10 Holmes, Michael, John Ameliorations apportees a ou concernant des agents contrastants
WO1993000933A1 (fr) * 1991-07-05 1993-01-21 University Of Rochester Particules poreuses non agregees ultra-petites piegant des bulles gazeuses
WO1993013808A2 (fr) * 1992-01-09 1993-07-22 Holmes, Michael, John Perfectionnements apportes aux agents de contraste
WO1995003835A1 (fr) * 1993-07-30 1995-02-09 Alliance Pharmaceutical Corp. Compositions a mocrobulles stabilisees pour applications mettant en ×uvre des ultrasons
US5605673A (en) * 1993-07-30 1997-02-25 Alliance Pharmaceutical Corp. Stabilized microbubble compositions for ultrasound
WO1995021631A1 (fr) * 1994-02-15 1995-08-17 Nycomed Imaging As Perfectionnements relatifs a des agents de contraste
WO1996026746A1 (fr) * 1995-02-28 1996-09-06 Alliance Pharmaceutical Corp. Emulsions gazeuses stabilisees contenant des phospholipides renforçant les contrastes dans les techniques d'imagerie par ultrasons

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20240226418A1 (en) * 2021-01-26 2024-07-11 Agitated Solutions Inc. Syringe-Based Microbubble Generator

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DE19602930A1 (de) 1997-07-24
WO1997026016A3 (fr) 1997-10-23
EP0874644A2 (fr) 1998-11-04
ZA97414B (en) 1997-07-17
CA2243174A1 (fr) 1997-07-24
AU1592597A (en) 1997-08-11
JP2000504317A (ja) 2000-04-11
NO983332D0 (no) 1998-07-17
NO983332L (no) 1998-07-17

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