DK165622B - MICROPARTICLE AND GAS BUBBLE CONTAINING ULTRA SOUND CONTROLLING AGENT AND EQUIPMENT AND PROCEDURE FOR ITS MANUFACTURING - Google Patents

Abstract

1. Contrast medium containing microparticles and gas bubbles for ultrasound diagnostics, characterised in that it contains microparticles of a mixture of a semi-solid or liquid surface-active substance and a non-surface-active solid in a liquid carrier.

Description

in DK 165622 B

The invention relates to a contrast agent for ultrasound diagnostics and to a device and method for producing the contrast agent.

The invention relates to the objects characterized in the claims.

The examination of organs with ultrasound (sonography) is a well established and practiced diagnostic method for 5 some years. Ultrasonic waves in the MHz range (above 2 MHz with wavelengths between 1 and 0.2 mm) are reflected at interfaces of different tissue types. The resulting echoes are amplified and made visible.

Of particular importance here is the study of the heart with this method, which is called echocardiography (J.I. Haft et al., Clinical Echocardiography, Futura, Mount Kisco, New York 1978, E.

Kohler, Clinical Echocardiography, Enke, Stuttgart 1979, G. Stefan et al., Echocardiography, Thieme, Stuttgart-New York 1981, G. Biamino, L. Lange, Echocardiography, Hoechst AG, 1983).

Since fluids - including blood - only provide ultrasound contrast when there are differences in density relative to the surroundings, opportunities were made to make the blood and its flow visible for an ultrasound examination, which is also possible by the addition of fine gas bubbles.

From the literature, several methods for making and stabilizing gas bubbles are known. For example, they can be produce by vigorously shaking or stirring solutions, such as saline, dye solutions, or pre-drawn blood, prior to injection into the blood stream.

Thus, while achieving ultrasound contrast giving, these methods are associated with weighty drawbacks that manifest themselves with poor reproducibility, strongly fluctuating size of the gas bubbles, and - due to some visible large bubbles - some risk of blood clots. These disadvantages were partially overcome by other methods of preparation such as through U.S. Patent No. 3,640,271, in which bubbles of reproducible size are produced by filtration or by the use of a direct current electrode apparatus. The advantage of being able to produce gas bubbles of reproducible size faces the disadvantage

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2 of the significant technical costs.

U.S. Patent No. 4,276,885 describes the preparation of defined size gas bubbles surrounded by a protective gelatin sheath of a prior to confluence. The finished bubbles can only be stored in a "frozen" state, e.g. when stored at refrigerator temperature, whereby they must be brought back to body temperature for use.

10 U.S. Patent No. 4,265,251 describes the preparation and use of solid saccharide sheath gas bubbles which may be filled with a pressurized gas. If under normal pressure, they can be used as an ultrasonic contrast agent; when used with elevated internal pressure, they serve blood pressure measurement.

15 Although the storage of the solid gas bubbles does not present a problem, the technical cost of manufacture is a significant cost factor.

The risks of the 20 contrast agents available at the state of the art are caused by two factors: the size and number of both the solid particles and the gas bubbles.

The state of the art so far illustrated allows the production of ultrasound contrast agents which always possess only 25 of the required properties: 1) exclusion of risk of blood clot, gas bubbles (size and number) - solid particles (size and number) 30 2) reproducibility 3) 4) can enter the lungs, e.g. 5) can enter the capillaries 35 6) sterility and pyrogenic freedom during preparation 7) easy manufacturability with reasonable cost supply, and 8) trouble-free storage.

DK 165622B

3 European Patent Application Publication No. 52575 discloses, however, the manufacture of gas bubbles which should possess these necessary properties. For the preparation of these gas bubbles, microparticles of a solid crystalline substance, such as e.g. galactose, in a carrier liquid, whereby the gas adsorbed to the particle surface or enclosed in voids between the particles or in intercrystalline voids forms the gas bubbles. The suspension of gas bubbles and microparticles thus formed is injected within 10 to 10 minutes. Although in European Patent Specification No. 52575 it is claimed that the suspension prepared by the method described is suitable to appear after injection into a peripheral vein both in the right side of the heart and after passage of the lung into the left side of the the heart and there make the blood and its flow visible by ultrasound examination, so this claim does not withstand a test. Thus, it was determined that according to the method described in European Application No. 52575, contrast agent injected into a peripheral vein did not produce ultra-20 echo echoes in the left side of the heart.

The object of the present invention was to provide an ultrasound diagnostic contrast agent capable of rendering the blood and its flow conditions visible for ultrasound not only on the right side of the heart, but also after the passage of the capillary layer of the lung. the left side of the heart. In addition, it should also allow the production of the bleeding of other organs, such as the myocardium, liver, spleen, and kidneys.

30

The novel agents according to claims 1-6 possess all the properties expected of such a contrast agent and which are discussed on page 2.

The present invention relates to a body-type ultrasonic and gas bubble-containing contrast agent for ultrasound diagnostics, which is characterized in that it contains microparticles of

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4 shows a mixture of a semi-solid or liquid interface active substance and a non-interface active solid in a liquid carrier, the liquid carrier being water or a physiological saline solution, the interface active substance being present in an amount of 0.01 to 5% by weight. and is a C4-C20 fatty acid, butyl stearate, polyethylene glycol sorbitan monostearate, sorbitan mono-1 aurate, polyethylene glycol-40 monolaurate, soybean oil sucrose glyceride, palm oil xylite or glycerine polyethylene glycol azine oleate, and a non-green substance of % by weight and is selected from galactose, lactose, sodium chloride and α-cyclodextrin.

Surprisingly, it was found that by suspending microparticles of a mixture of a semi-solid or liquid interface surfactant and a non-interface active solid in a carrier liquid, an ultrasonic contrast agent is obtained which, after injection into a peripheral vein, also allows reproducible ultrasound recordings. blood in the arterial left side of the heart.

Since the left side of the heart can be reached with the ultrasound contrast agent of the invention after intravenous insertion, ultrasound contrasts of other of the blood-aortic organs are also possible after venous insertion, such as the myocardium, liver, spleen, and kidneys, and others. It is understood that the ultrasonic contrast agent of the invention is also suitable for contrasts in the right side of the heart and for all other applications as ultrasound contrast agent.

The semi-solid or liquid interface surfactants which are constituents of the mixture necessary for the preparation of the microparticles are all physiologically compatible in the amounts used, i.e. they have low toxicity and / or are biodegradable and their melting point is lower than room temperature, ie. that they are semi-solid or liquid at room temperature. The interface active agents are selected from C4-35 C20 fatty acids, butyl stearate, polyethylene glycol sorbitan monostearate, sorbitan monolaurate, polyethylene glycol-40 monolaurate,

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5 isole oil sucrose glyceride, palm oil xylite or glycerine polyethylene glycol rizinoleate, with butyl stearate, soya oil sucrose segyceride, polyethylene glycol sorbitan monostearate and palm oil xylite being preferred.

5

The interface active substance is used at a concentration of 0.01 to 5% by weight, preferably 0.04 to 0.5% by weight.

Sodium chloride, galactose, lactose and γ-cyclodex steps, with galactose, lactose and α-cyclodextrin being the preferred ones, are considered as non-surfactant constituents of the mixture required for the preparation of the microparticles. They are present at a concentration of 5 to 50% by weight, preferably 9 to 40% by weight, in the composition of the invention.

To prepare the microparticles, the non-surfactant is recrystallized under sterile conditions. Then the interface active substance is mixed and comminuted with the non-interface active solid under sterile conditions, e.g. by grinding in an air jet mill until the desired particle size is reached. A particle size of less than 1-50 μιη is obtained, preferably 1-10 μπι. The particle size is determined in suitable measuring devices. The weight-to-weight ratio of interface active agent to non-interface active agent can range from 0.2-100: 100.

Both the microparticle size obtained by the comminution process as well as the size of the gas bubbles contained in the contrast agent 30 of the invention ensures a harmless passage of capillary leather and of the lung capillary layer and precludes the formation of blood clots.

The gas bubbles required for contrasting are partially transported by the suspended microparticles, absorbed to the surface of the microparticles, or enclosed in the voids between the microparticles or intercrystalline.

The volume of gas transported by the microparticles in the form of gas bubbles amounts to 0.02 - 0.6 ml / g microparticles.

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6

Next to the transport function for the task 5, the shaving fluid has to stabilize the suspension of microparticles and gas bubbles, ie. preventing the sedimentation of the microparticles and confluence of the gas bubbles and delaying the dissolution process of the micro-particles.

10 As a liquid carrier, water and physiological saline are discussed.

The invention also relates to a process for the preparation of the present invention according to claim 8.

15

The process of the invention is characterized by combining microparticles of a mixture of a physiologically compatible semi-solid or liquid interface active agent in an amount of 0.01 to 5% by weight, which interface active agent is selected from a C4-C20 fatty acid, butyl stearate. , polyethylene glycol colsorbitan monostearate, sorbitan monolaurate, polyethylene glycol 40 monolaurate, soybean oil sucrose glyceride, palm oil xylite or glycerine polyethylene glycol rizinoleate, and a physiologically compatible non-surfactant, which is a 50% by weight, non-interfacial, in an amount of from 5 to 25 sodium chloride and α-cyclodextrin, in a physiologically compatible carrier liquid selected from water and physiological saline solution and shake this until a homogeneous suspension occurs.

To prepare the ready-to-use ultrasonic contrast agent, the sterile carrier liquid is added to those of a mixture of a semi-solid or liquid interface active and a non-interface active solid and microparticles shaken until a homogeneous suspension has formed, to which approx. The resulting suspension is injected immediately after preparation and, not later than 5 minutes after, as a total amount into a peripheral vein or into a catheter already present, from 0.01 ml to 1 ml / kg body weight.

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7

For convenience, the components necessary for the preparation of the agent of the invention, such as carrier liquid (A) and microparticles, of a mixture of a semi-solid or liquid interface active substance and a non-interface active solid (B) are stored sterile in one for one. Examine required amount in two separate containers. Both containers have closures that allow removal and addition by means of a syringe under sterile conditions (small vials). The size of container B must be such that the contents of container A can be transferred to B by means of a syringe and the combined components can be shaken. The invention therefore also relates to an equipment according to claim

The equipment is peculiar in that it consists of 15 of βη container containing a volume of 5-10 ml, provided with a closure enabling the contents to be withdrawn under sterile conditions containing 4 ml of liquid carrier selected from water and physiological saline solution, and b) of another container having a volume of 5-10 ml, provided with a closure which allows withdrawal of the contents or addition of a substance mixture under sterile conditions containing microparticles of a mixture of a semi-solid or liquid interface active agent selected from a C4 ~ C20 ~ fatty acid, butyl stearate, polyethylene glycol sorbitan monostearate, sorbitan monolaurate, polyethylene glycol-40 monolaurate, soybean oil sucrose glyceride, palm oil xylite and glycerine polyethylene glycolrizinoleate in an amount of 0.01 to 5 wt. solid selected from galactose, lactose, sodium chloride and α-cyclodextrin with an average particle size of less e than 1 to 10 30 µm in an amount of from 5 to 50% by weight, preferably from 9 to 40% by weight, based on the total weight of the agent, wherein the weight ratio of interface active agent to non-interface active agent is 0.02-100: 100.

35

The conduct of an echocardiographic examination of a 10 kg heavy baboon should demonstrate the use of the contrast agent of the invention:

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8 5 5 ml of carrier liquid (prepared according to Example 1 Ά) are taken from a medicine bottle with a syringe and added to 2 g of microparticles (made according to Example 1 B) which are in another medicine bottle and shaken for approx. 5-10 seconds until a homogeneous suspension has formed. 10 ml of this suspension is injected into a peripheral vein (V. jugularis, brachialis or saphena) over a three-way cock with an infusion rate of at least 1 ml / sec, and better at 2-3 ml / sec.

Immediately after the contrast agent injection, the injection of 10 ml of physiological saline solution follows at the same rate, so that the contrast agent lump is maintained as far as possible until it reaches the right side of the heart. Before, during and after the injection, a commercially available echocardiography head is held against the test animal's chest so that a typical cross-section is obtained through the right and left sides of the heart. This experimental device corresponds to the state of the art and is known to those skilled in the art.

When the ultrasound contrast agent reaches the right side of the heart, in the 2-D echo image or in the M-mode echo image, you can follow how the contrast-marked blood first reaches the level of the right ventricle, then the right ventricle and pulmonary artery, whereby for approx. 10 sec. a homogeneous filling occurs. While the cavities in the right side of the heart are emptied again, contrast-marked blood reappears after the pulmonary passage in the pulmonary veins, homogeneously fills the left ventricle, left ventricle and aorta, where contrast persists 2-3 times longer than on the right side. of the heart.

In addition to the preparation of blood flow through the caves in the left side of the heart, it also produces a preparation 35 of the myocardium that reflects the bleeding.

However, the use of the ultrasound contrast agent of the invention is not limited to the visualization of blood flow in the arterial part of the heart after venous insertion, but the

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9 is also used with excellent results when examining the right side of the heart and other organs as a contrast agent.

Example 1.

5 A. Manufacture of carrier wash.

Water for injection is drained into 4 ml portions in 5 ml vials and sterilized for 20 minutes at 120 ° C.

10 B. Preparation of Microparticles.

Under sterile conditions, a sterile filtered solution of 0.5 g of butyl stearate in 40 g of isopropanol is applied to 199.5 g of sterile 15 galactose particles, evaporated at 40 ° C and 200 torr, and ground with an air jet mill until the following size distribution of particles:

Average 1.9 mm 20 min. 99% <6 mm min. 90% <3 m 2.

The determination of the particle size and their distribution takes place in a particle measuring apparatus, e.g. after suspending in isopro-25 panol. The microparticle pre-packing takes place in 5 ml medicine bottles with 2 g in each.

C. For the preparation of 5 ml ready-to-use ultrasonic contrast agent, the contents of a medicine bottle with carrier liquid (water 30 for injection, A) are added by means of a syringe to the medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed (5-10 SEC.).

Example 2.

35 A. Preparation of carrier liquid.

Water for injection is drained into 4 ml portions in 5 ml

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10 vials and sterilized for 20 minutes at 120 ° C.

B. Preparation of Microparticles.

Under sterile conditions, apply a sterile filtered solution of 0.5 g of soybean sucrose glyceride in 40 g of isopropanol to 199.5 g of sterile galactose particles, evaporate at 40 ° C and 200 torr and grind with an air jet mill until the following size distribution of the particles:

Average 1.9 Mm min. 99% <6 μη min. 90% <3 mm.

15

The determination of the particle size and their distribution takes place in a particle measuring apparatus, e.g. after suspending in isopropanol. The final packing of the microparticles takes place in 5 ml medicine bottles with 2 g in each.

20 C. To prepare 5 ml ready-to-use ultrasonic contrast agent, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to the medicine bottle with micropraticles (B) and shaken until a homogeneous suspension is formed (5-10 SEC.).

25

Example 3

A. Manufacture of Bar Wash.

30 g of sodium chloride are dissolved in water up to a volume of 500 ml, the solution is pressed through a 0.2 mm filter, 4 ml portions of this solution are drained into 5 ml medicine bottles and sterilized for 20 minutes at 120 ° C.

35 B Manufacture of microparticles.

Under sterile conditions, a sterile filtered solution of 0.5 g of polyethylene glycol sorbitan monostearate in 40 g of isopropanol is applied.

DK 165622B

11 on 199.5 sterile galactose particles, isopropanol is evaporated at 40 ° C and 200 torr and ground with an air jet mill until the following size distribution of the particles is achieved: 5

Mean 1.9 μιη min. 99% <6 μιη min. 90% <3 μιη.

The determination of the particle size and their distribution takes place in a particle measuring apparatus, e.g. after suspending in isopropanol. The final packing of the microparticles takes place in 5 ml medicine bottles with 2 g in each.

C. To prepare 5 ml of ready-to-use ultrasonic contrast agent, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to the medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed (5-10). SEC.).

Example 4

A. Preparation of carrier liquid.

25

Dissolve 4.5 sodium chloride in water to a volume of 500 ml, press the solution through a 0.2 mm filter, drain 4 ml portions of this solution into 5 ml medicine vials and sterilize for 20 minutes at 120 ° C.

B. Preparation of Microparticles.

Under sterile conditions, a sterile filtered solution of 0.5 g of palm oil xylite in 40 g of isopropanol on 199.5 g of 35 sterile galactose particles is applied, evaporated at 40 ° C and 200 torr, and ground with an air jet mill until the following size distribution of the particles: 12

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Mean 1.9 μΐη min. 99% <6mm - min. "90% <3mm.

The determination of the particle size and their distribution takes place in a particle measuring apparatus, e.g. after suspending in isopropanol. The final packing of microparticles takes place in 5 ml medicine bottles with 2 g in each.

10 c. For the preparation of 5 ml ready-to-use ultrasonic contrast agent, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to the medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed (5-10 SEC.).

15

Example 5

A. Manufacture of Bar Wash.

20 Water for injection is filled in 4 ml in 5 ml medicine vials and sterilized for 20 min. at 120 ° C.

B. Preparation of Microparticles.

Under sterile conditions, a sterile filtered solution of 0.5 g of glycerine polyethylene glycol rizinoleate in 40 g of isopropanol is applied to 199.5 g of sterile galactose particles, isopropanol is evaporated at 40 ° C and 200 Torr and ground with an air jet mill until obtained the following particle size distribution:

Average 1.9 pm min. 99% <6 pm min. 90% <3 pm.

The particle size and their distribution are determined in a particle measuring apparatus, e.g. after suspending in isopropa- 35

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13 nol. The final packing of the microparticles takes place in 5 ml medicine bottles with 2 g in each.

v C. For the preparation of 5 ml ready-to-use ultrasonic contrast agent 5, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to the medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed (5 -10 sec.).

Example 6.

A. Preparation of carrier fluids.

Water for injection is filled into 4 ml portions in 5 ml medicine vials and sterilized for 20 min. at 120 ° C.

B. Preparation of Microparticles.

Under sterile conditions, apply a sterilized solution of 0.5 g of sorbitan monolaurate in 40 g of isopropanol on 199.5 g of sterile galactose particles, evaporate at 40 ° C and 200 Torr and grind with an air jet mill until obtained the following particle size distribution: 25 Mean 1.9 µm min. 99% <6 pm min. 90% <3 pm

The particle size and their distribution are determined in a particle measuring apparatus, e.g. after suspending in isopropanol. The final packing of the microparticles takes place in 5 ml medicine bottles with 2 g in each.

C. For the preparation of 5 ml ready-to-use ultrasonic contrast agent 35, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to a medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed (5 -10 sec.).

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14

Example 7

A. Manufacture of carrier wash.

4.5 g of sodium chloride are dissolved in water to a volume of 500 ml, then the solution is pressed through a 0.2 mm filter and 4 ml portions of this solution are filled into 5 ml of medicine bottles and sterilized for 20 minutes. at 120 ° C.

10 B. Preparation of Microparticles.

Under sterile conditions, apply a sterile filtered solution of 0.5 g of polyethylene glycol-40 monolaurate in 40 g of isopropanol on 199.5 g of sterile lactose particles, evaporate 15 at 40 ° C and 200 Torr, and grind with a 1 Uft jet mill 1 e until the following particle size distribution is obtained:

Average 1.9 pm min. 99% <8 pm 20 min. 90% <5 pm.

The particle size and their distribution are determined in a particle measuring apparatus, e.g. after suspending in isopropanol. The final packing of the microparticles takes place in 5 ml medicine bottles with 2 g in each.

C. For the preparation of 5 ml ready-to-use ultrasonic contrast agent, the contents of a medicine bottle with carrier liquid (water for injection, A) are added by means of a syringe to the medicine bottle with microparticles (B) and shaken until a homogeneous suspension is formed ( 5-10 sec.).

Example 8.

35 a. Preparation of carrier liquid.

4.5 g of sodium chloride are dissolved in water to a volume of 500 ml, then the solution is pressed through a 0.2 mm filter, and

Claims (8)

1. Microparticle and gas bubble containing contrast agent for ultrasound diagnostics, characterized in that it contains microparticles of a mixture of a semi-solid or liquid interface active substance and a non-interface active solid in a liquid carrier, the liquid carrier being water or a physiological solution. , the interface active agent is present in an amount of 0.01 to 5% by weight and is a C4-DK 165622B c20-fatty acid, butyl stearate, polyethylene glycol sorbitan monostearate, sorbitan monolaurate, polyethylene Tyco 1-40 monolaurate, soyaol fesaccharose glyceride, palm oil xylite or glycerine polyethylene glycol rizinoleate, and the non-interface active substance is present in an amount of from 5 to 50% by weight and is galactose, lactose, sodium chloride or α-cyclodextri η. An agent according to claim 1, characterized in that it contains microparticles containing as semi-solid or liquid interface active substance, butyl stearate, soya oil sucrose glyceride, polyethylene glycol sorbitan monostearate or palm oil iexyl it in a concentration of 0.01 to 5% by weight, preferably 0, 04 to 1% by weight. An agent according to claim 1, characterized in that it contains microparticles of a mixture of butyl stearate and galactose in water. Agent according to claim 1, characterized in that it contains microparticles of a mixture of soybean oil sucrose glyceride and galactose in water. Agent according to claim 1, characterized in that it contains microparticles of a mixture of polyethylene glycol sorbitan monostearate and galactose in a physiological boiling salt solution. An agent according to claim 1, characterized in that it contains microparticles of a mixture of palm oil xylitol and 30 galactose in a physiological saline solution. Apparatus for producing a microparticle and gas bubble containing ultrasonic contrast agent according to claim 1, characterized in that it consists of a) a container having a volume of 5-10 ml, provided with a closure for withdrawing the contents below sterile conditions containing 4 'ml of liquid carrier selected from water and physiological saline solution, and b) in another container having a volume of 5-10 ml, provided with a closure which allows the contents to be removed or 1 - setting a substance mixture under sterile conditions containing microparticles of a mixture of a semi-solid or liquid interface active substance selected from a C4-C20 fatty acid, butyl stearate, polyethylene glycol sorbitan monostearate, sorbitan monolaurate, polyethylene glycol-40 10 seglyceride, palm oil xylit and glycerine polyethylene glycol rizinoleate in an amount of 0.01 to 5% by weight, based on the total weight of the agent, and a non-interface active solid selected from galactose, lactose, sodium chloride and α-cyclodextrin having an average particle size of less than 1 to 10 µm in an amount from 5 to 50% by weight, preferably from 9 to 40% by weight, based on the total weight of the agent, wherein the weight ratio of interface active substance to the existing non-interface active substance is 0.02-100: 100. A method of preparing a microparticle and gas-containing ultrasonic diagnostic contrast agent according to claim 1, characterized in that microparticles are combined by a mixture of a physiologically compatible semi-solid or liquid interface active agent in an amount of 0.01 to 25% by weight. which interface active agent is selected from a C 5 to 50% by weight, which non-interface active substance is selected from galactose, lactose, sodium chloride and α-cyclodextrin, in a physiologically compatible carrier liquid selected from water and physiologically saline solution and shake this until a homogeneous suspension is obtained.