JP2003048826A - Perfluorocarbon emulsion and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an emulsion which comprises a perfluorocarbon compound as an active ingredient, has excellent stability, is fine and can be sterilized by filtration, and to provide a method for producing the same. SOLUTION: This perfluorocarbon emulsion is characterized by comprising (A) 10-80 wt.% of the perfluorocarbon compound, (C) 0.1-10 wt.% of a medically acceptable emulsifying agent, (D) a water phase component and (B) 0.1-90 wt.% based on the component (A) of a compound represented by general formula (1) R<1> -X-R<2> (1) (R<1> is a 6-12C alkyl group containing a >=6C perfluoroalkyl part; R<2> is a 12-22C alkyl group; X is a specific ester or ether bond).

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a perfluorocarbon emulsion and a method for preparing the same. More specifically, a perfluorocarbon compound, an ester or ether having a perfluoroalkyl group and an alkyl group in the molecule, an emulsifier, and an oxygen carrier, which comprises an aqueous phase component,
The present invention relates to a perfluorocarbon emulsion useful as an organ preservation solution or a contrast agent, and a method for producing the same.

[0002]

2. Description of the Related Art Perfluorocarbon compounds are capable of dissolving and transporting a large amount of oxygen even though they are chemically inert. Therefore, they are oxygen carriers, organ preservation solutions, or physical therapies such as ultrasonic waves and X-rays. It is expected as an active ingredient such as a contrast agent. In these medical applications, it is necessary to stably atomize the above-mentioned perfluorocarbon compound, and therefore, it is generally prepared as an emulsion. As such a perfluorocarbon emulsion,
For example, “Fluosol DA 20%” (Midori Cross Co., Ltd.) is widely known. This consists of two types of perfluorocarbon compounds, namely perfluorodecalin and perfluorotripropylamine, and two types of surfactants, egg yolk phospholipids and polyoxyethylene polyoxypropylene glycol (Pluronic F-68, BASF). It is a formulation that is emulsified and dispersed in water using a mixture of However, since this emulsion is unstable in a liquid state, it has been problematic for use in medical applications. In general, since these perfluorocarbon compounds have strong water repellency, it is very difficult to stably emulsify and disperse them in water. Therefore, much work has been done to prepare stable emulsions.

For example, Japanese Patent Publication No. 1-281212 and Japanese Patent Laid-Open No. 3-72423 disclose emulsions using a cyclic perfluoronitrogen-containing compound as a perfluorocarbon compound. These were intended to obtain highly stable emulsions with a general composition by adopting a specific perfluorocarbon nitrogen-containing compound as an active ingredient.
It had to be stored at a low temperature of ℃. In addition, JP-A-4-1
Japanese Patent No. 54718 discloses a perfluorocarbon emulsion obtained by mixing a composition comprising a perfluorocarbon, a phospholipid and an aqueous solvent, which is emulsified by using the three components, and a liquid containing a high molecular nonionic surfactant. ing. This is a highly stable emulsion by mixing an emulsified composition consisting of perfluorocarbon, phospholipid and an aqueous solvent with a liquid containing a polymeric nonionic surfactant such as polyoxyethylene polyoxypropylene glycol. However, it is hard to say that it is sufficient in terms of long-term stability. In addition, Japanese Patent Fair 8-228
Japanese Patent No. 15 discloses an emulsion composed of a perfluorocarbon compound, an oil, a surfactant and water. This is one in which a perfluorocarbon compound is dispersed in oil, and the mixture is emulsified in water by a surfactant.However, the perfluorocarbon compound is practically hard to be mixed with oil because of its oil repellency, and in most cases Since the perfluorocarbon compound has a larger specific gravity than oil, it was still insufficient in stability.

[0004]

As described above, all of the conventional methods are insufficient in terms of particle size and stability as pharmaceutical emulsions, particularly as pharmaceutical preparations that are supposed to be administered intravenously. It was not practical. Particularly in medical applications, emulsions are difficult to sterilize by filtration, and therefore, they are required to be heat-sterilized as much as possible, but not only ordinary storage stability but also stability after heat-sterilization is maintained. It was extremely difficult to prepare a perfluorocarbon emulsion. In particular, although perfluorohydrocarbons are excellent in oxygen transportability and half-life in vivo, it is possible to prepare fine and stable emulsions due to their molecular structure and polarity. Since it was difficult, even when only perfluorohydrocarbon was selected as the active ingredient, a method for preparing a fine and highly stable emulsion was eagerly desired. Further, in such an emulsion, there has been a strong demand for an emulsion in which filtration sterilization can be selected as necessary, instead of heat sterilization which may reduce the stability. The present invention is intended to solve the above problems. That is, the object of the present invention, a perfluorocarbon compound as an active ingredient, excellent stability,
Another object of the present invention is to provide a fine emulsion which can be sterilized by filtration and a method for producing the emulsion.

[0005]

Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventors have found that in an emulsion containing a perfluorocarbon compound as an active ingredient, a specific perfluoroalkyl group and alkyl By generating a specific amount of a specific ester or ether having a group, and further by using a high pressure jet flow emulsifier, they are generated between liquids of two jet streams facing each other under specific high pressure conditions. It was found that an emulsion having excellent stability can be prepared by emulsifying by using shearing force, and that the emulsion prepared by this method has an extremely fine particle size and can be sterilized by filtration. The invention was completed.

That is, the present invention is as follows. (1) A perfluorocarbon emulsion comprising (A) 10 to 80% by weight of a perfluorocarbon compound, (C) 0.1 to 10% by weight of a medically acceptable emulsifier, and (D) an aqueous phase component, , (B) a compound represented by the following general formula (1) in an amount of 0.1 to 9 relative to the component (A).
A perfluorocarbon emulsion containing 0% by weight. R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents

[0007]

[Chemical 4]

Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. (2) The compound (B) represented by the general formula (1) is 40 ° C.
The perfluorocarbon emulsion according to (1) above, which is liquid at the temperature of. (3) The perfluorocarbon emulsion according to (1) above, wherein the medically acceptable emulsifier (C) contains a phospholipid. (4) The perfluorocarbon emulsion according to (1) above, wherein the medically acceptable emulsifier (C) is a mixture of a phospholipid and a nonionic surfactant in a ratio of 10:90 to 99: 1 (weight ratio). (5) The perfluorocarbon emulsion according to the above (3) or (4), wherein the phospholipid constituting the medically acceptable emulsifier (C) contains a phospholipid having a polyethylene glycol chain. (6) The molecular weight of the polyethylene glycol chain in the phospholipid having a polyethylene glycol chain is 400 to 20.
The perfluorocarbon emulsion according to the above (5), which is 000. (7) (A) Perfluorocarbon compound 10 to 80% by weight, (B) 0.1 to 90% by weight with respect to the component (A)
Of a compound represented by the following general formula (1), (C) a medically acceptable emulsifier of 0.1 to 10% by weight, and (D) an aqueous phase component mixed solution under a jet pressure of 150 MPa or more. As an emulsion, and the jet stream is made to collide with a wall surface arranged facing the jet port, and the jet stream is inverted in a direction substantially opposite to the direction of the jet stream to emulsify the perfluorocarbon emulsion. Manufacturing method. R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents

[0009]

[Chemical 5]

Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. (8) (A) Perfluorocarbon compound 10 to 80% by weight, (B) 0.1 to 90% by weight with respect to the component (A)
A method for producing a perfluorocarbon emulsion comprising a compound represented by the following general formula (1), (C) 0.1 to 10% by weight of a medically acceptable emulsifier, and (D) an aqueous phase component: First, a mixture of (C) a medically acceptable emulsifier and (D) an aqueous phase component is jetted as a jet stream under a pressure of 50 to 250 MPa, and in the jet stream,
After preliminarily emulsifying by injecting a mixture of (A) a perfluorocarbon compound and (B) a compound represented by the general formula (1), the obtained preliminarily emulsified liquid was mixed with 150M.
It is jetted as a jet stream under a pressure of Pa or higher, and the jet stream is collided with a wall surface facing the jet port,
A method for producing a perfluorocarbon emulsion, which comprises emulsifying by reversing in a direction substantially opposite to the direction of the jet flow. R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents

[0011]

[Chemical 6] Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. (9) A perfluorocarbon emulsion prepared by the method according to (7) or (8), wherein the average particle size of the dispersed phase is 200 nm or less.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in more detail below. According to the purpose of the present invention, the perfluorocarbon compound which is the component (A) used in the perfluorocarbon emulsion of the present invention has oxygen-carrying ability and has a low harmful reaction to the living body. It can be appropriately selected from the above. Examples of such perfluorocarbon compounds include perfluorohydrocarbons, perfluoroalkyl bromides, perfluoroalkylamines, and the like. In the present invention, the above general formula (1)
By further emulsifying the compound represented by and emulsifying by using a shearing force by a high-pressure jet flow, a fine and stable perfluorocarbon emulsion can be obtained. Not only high perfluoroalkylamines but also low polar perfluorocarbon compounds, which were difficult to be stably emulsified by conventional techniques, can be made into emulsions excellent in stability.

Therefore, examples of the perfluorocarbon compound as the component (A) in the present invention include perfluorocycloalkanes such as perfluorocyclohexane, perfluoroalkylcycloalkanes such as perfluorotrimethylcyclohexane and perfluoroisopropylcyclohexane, and perfluoro. One or two compounds from perfluoroalkyldecalin such as methyldecalin, perfluorodecalin, perfluoroalkane, perfluorooctyl bromide, perfluorodecyl bromide, etc., which have a relatively short half-life in vivo and are highly safe The above may be used in combination. The content of the above-mentioned perfluorocarbon compound is 10 to 80% by weight, preferably 20 to 70% by weight in the perfluorocarbon emulsion.
It is adjusted to be within the range of weight%. If the amount used exceeds 80% by weight, the viscosity of the finally prepared emulsion may be increased and the stability thereof may be decreased. It is not preferable because the performance is reduced.

In the method for preparing a perfluorocarbon emulsion of the present invention, in addition to the perfluorocarbon compound (component A), the medically acceptable emulsifier (component C), and the aqueous phase component (component D), the addition of (B ) As an ingredient,
A compound represented by the following general formula (1) having one or more selected from esters or ethers having a specific perfluoroalkyl group and an alkyl group in the molecule is added to the component (A). By adding 0.1 to 90% by weight, an emulsion composed of fine particles and stable can be prepared. R ¹ —X—R ² (1) In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 12 carbon atoms.
~ 22 alkyl groups. X represents that R ¹ and R ² are ester or ether bond,
For example, in the case of an ester, X is

[0015]

[Chemical 7] And in the case of ether, X ³ : -O-. In addition, other ester or ether bonding modes include

[0016]

[Chemical 8] (In the formula, R ³ is an alkylene group having 2 to 8 carbon atoms.)
Such as R ¹ on each side of the alkanedicarboxylic acid
Or R ² is an ester bond, or

[0017]

[Chemical 9]

Or X ⁶ : -O-R ³ -O- (in the formula, R ³ is the same as in the case of X ⁴ above), R ¹ and R ² are an ester or R ^{2 on} both sides of the alkanediol. The thing etc. which were ether-bonded are mentioned. The component (B) used in the method for producing a perfluorocarbon emulsion of the present invention is an alkyl group having 6 to 12 carbon atoms (R ¹ ) having a perfluoroalkyl moiety having 6 or more carbon atoms.
And one or more compounds selected from the compounds represented by the general formula (1) having an alkyl group (R ² ) having 12 to 22 carbon atoms can be appropriately selected and used. The alkyl group (R ¹ ) may partially have a hydrocarbon group as long as the carbon chain consisting of carbon and fluorine has at least 6 carbon atoms. At this time, if the chain length (carbon number) of the alkyl group (R ¹ ) is smaller than 6, the effect of adding the component (B) cannot be sufficiently obtained, and conversely if the chain length (carbon number) exceeds 12, Since the melting point of the component (B) becomes high, the stability of the finally obtained perfluorocarbon emulsion may decrease.

Also in the case of the alkyl group (R ² ), similarly, when the chain length (carbon number) is smaller than 12, (B)
The effect of adding the components is not sufficient, and conversely the chain length (carbon number)
When the value exceeds 22, the melting point as the component (B) becomes high and the compatibility with the perfluorocarbon compound becomes low, so that the stability of the resulting perfluorocarbon emulsion may be lowered. Therefore, as the component (B) used in the method for preparing a perfluorocarbon emulsion of the present invention, an alkyl group having 6 to 12 carbon atoms (R ¹ ) having a perfluoroalkyl moiety having 6 or more carbon atoms and 12 to 12 carbon atoms are used. A compound having 22 alkyl groups (R ² ) which is preferably liquid at 40 ° C. is desirable. The component (B) used in the present invention is not particularly limited as long as it is represented by the above general formula (1), but it is available and economical, and the component (B) can be easily purified or synthesized. In consideration of generation of impurities due to side reaction at the time, R ¹ − in general formula (1)
The compound represented by X ¹ -R ² is preferably selected.

Examples of such a compound include, for example, 6 to 6 carbon atoms having a perfluoroalkyl moiety having 6 or more carbon atoms.
From the esters of 12 primary alcohols and fatty acids having 12 to 22 carbon atoms, one kind or two or more kinds can be appropriately selected and used. Considering the stability of the finally obtained perfluorocarbon emulsion, 40
Those which are liquid at 0 ° C. are preferably used, and examples of such a compound include primary alcohols having 6 to 10 carbon atoms and primary alcohols having 6 to 8 carbon atoms and 16 to 2 carbon atoms.
Esters obtained with 0 unsaturated fatty acids are preferably selected. Preferable specific examples of the above-mentioned ester include, for example, palmitoleic acid, oleic acid, linoleic acid, linolenic acid, gondoinic acid, erucic acid, etc.
~ 22 and fatty acids having an unsaturated alkyl group in the molecule, and primary carbon atoms having 6-12 carbon atoms, such as (perfluorohexyl) ethanol and (perfluorooctyl) ethanol, having 6 or more carbon atoms Examples thereof include esters with alcohol.

In the present invention, the amount of the component (B) used is
0.1 with respect to the perfluorocarbon compound (A)
It is adjusted to be in the range of 90 to 90% by weight, preferably 0.5 to 60% by weight, and more preferably 1 to 50% by weight. At this time, if the amount used exceeds 90% by weight,
Not only the oxygen carrying capacity of the finally prepared perfluorocarbon emulsion becomes low, but also the compatibility with the component (B) may decrease depending on the molecular structure of the component (A), resulting in low stability. It is not preferable because it may occur. On the other hand, if it is less than 0.1% by weight, the effect of adding the component (B) cannot be sufficiently obtained, and as a result, the stability of the emulsion may be lowered, which is not preferable. Therefore, it is desirable that the content of the component (B) is appropriately selected and used within the above range, depending on the selected perfluorocarbon compound, in such an amount as to be dissolved therein.

In the present invention, the medically acceptable emulsifying agent which is the component (C) can be selected from the emulsifying agents generally used in pharmaceutical applications. Among them, those containing phospholipids or those not containing phospholipids are used. A combination of 10:90 to 99: 1 by weight ratio with the ionic surfactant is preferably selected. Among them, phospholipids are particularly preferably selected and used in the emulsifier (C) in an amount of preferably 5 to 100% by weight, more preferably 10 to 100% by weight. Examples of the phospholipid used as the component (C) include natural phospholipids such as egg yolk phospholipids and soybean phospholipids, hydrogenated phospholipids obtained by hydrogenating these natural phospholipids, and C12-22 carbon atoms. And synthetic phospholipids having an acyl group introduced therein. Suitable specific examples of such phospholipids include natural phospholipids such as egg yolk phosphatidylcholine and soybean phosphatidylcholine, hydrogenated phospholipids such as hydrogenated egg yolk phosphatidylcholine and hydrogenated soybean phosphatidylcholine, dimyristoylphosphatidylcholine, dimyristoylphosphatidylethanolamine, dipalmitoyl. Phosphatidylcholine, dipalmitoylphosphatidylethanolamine, distearoylphosphatidylcholine, distearoylphosphatidylethanolamine, dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine, 1-
Examples thereof include synthetic phospholipids such as palmitoyl-2-oleoylphosphatidylcholine.

Among them, considering the stability of the emulsion to be prepared, natural or synthetic phosphatidylcholine is preferably used, and particularly naturally-derived diacylphosphatidylcholine egg yolk phosphatidylcholine or hydrogenated egg yolk phosphatidylcholine is preferably selected.
In addition, the component (C) used in the method for producing a perfluorocarbon emulsion of the present invention contains various acyl groups other than the above phospholipids as long as the stability of the finally prepared emulsion is not affected. Phosphatidic acid having
Phosphatidylinositol, phosphatidylglycerol, phosphatidylserine, may contain phospholipids such as sphingomyelin, further, in the range that does not affect the stability and hemolysis of the finally obtained emulsion,
The phospholipid may contain a fatty acid salt such as lysophospholipid or sodium oleate.

Examples of the nonionic surfactant used in combination with the phospholipid as the component (C) include polyoxyethylene polyoxypropylene glycol, polyoxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid ester and the like. Among them, polyoxyethylene polyoxypropylene glycol is preferably used. The nonionic surfactant is
The weight ratio of phospholipid and nonionic surfactant is 10: 9.
It is preferably used so as to be 0 to 99: 1.
The amount of the component (C) used in the present invention is adjusted so as to be within the range of 0.1 to 10% by weight, preferably 0.5 to 5% by weight, in the perfluorocarbon emulsion. At this time, if the amount used is less than 0.1% by weight, the effect as an emulsifier is not obtained, the stability of the finally prepared emulsion is reduced, and the emulsion may be separated as a result. On the other hand, if it exceeds 10% by weight, the viscosity of the emulsion may increase, resulting in a decrease in the fluidity of the emulsion, which is not preferable.

In the present invention, the stability of the finally prepared perfluorocarbon emulsion is improved by incorporating a phospholipid having a polyethylene glycol chain into the emulsifier (C) in addition to the phospholipid. It can be further improved. In the present invention, a phospholipid having a polyethylene glycol chain (hereinafter referred to as a PEGylated phospholipid) contained in the emulsifier (C) has a structure in which polyethylene glycol is bonded to a hydrophilic group of the phospholipid. The end of the polyethylene glycol chain on the side not bound to the phospholipid may be a hydroxyl group as it is, or the terminal hydroxyl group may be modified with a methyl group, an ethyl group or the like. In the PEGylated phospholipid, the phospholipid that binds the polyethylene glycol chain is not particularly limited as long as it is a phospholipid, but it is relatively easy to introduce the polyethylene glycol chain and the degree of hydrolysis after introduction is low. Is preferable, and phosphatidylethanolamine is preferably selected (the phosphatidylethanolamine having a polyethylene glycol chain is hereinafter referred to as PEGylated phosphatidylethanolamine).

In the present invention, PEGylated phosphatidylethanolamine means egg yolk phospholipid or soybean phospholipid,
Alternatively, it can be obtained by using a phosphatidylethanolamine contained in the hydrogenated phospholipids thereof, a diacylphosphatidylethanolamine prepared as a synthetic phospholipid or the like, and binding polyethylene glycol or a derivative thereof thereto. In the present invention, as a method for binding phosphatidylethanolamine and polyethylene glycol or a derivative thereof,
Examples thereof include a method using cyanuric chloride, a method using carbodiimide, a method using glutaraldehyde, and a method using carbamate. Among them, if the method using carbodiimide is used, for example, monomethoxypolyethylene glycol and succinic anhydride are reacted to introduce a carboxyl group (of succinic acid) into the hydroxyl group of monomethoxypolyethylene glycol, and then this and phosphatidylethanolamine Can be reacted in the presence of carbodiimide to prepare a PEGylated phosphatidylethanolamine having a polyethylene glycol chain introduced via an amide bond.

In the case of the method using carbamate, for example, monomethoxypolyethylene glycol and carbonyldiimidazole are reacted to obtain imidazolecarbamate of methoxypolyethylene glycol, and then this and phosphatidylethanolamine are added in the presence of triethylamine. By introducing a polyethylene glycol chain through a urethane bond by reacting with P
EGylated phosphatidyl ethanolamine can be prepared. In the present invention, the molecular weight of the polyethylene glycol chain in the phospholipid having a polyethylene glycol chain is preferably 400 to 20,000, more preferably 1,000 to 10,000, and particularly preferably 2,000.
~ 8000. If the molecular weight of the polyethylene glycol chain exceeds 20,000, the water solubility of the above-mentioned PEGylated phospholipid becomes high, and as a result, the stability of the prepared perfluorocarbon emulsion may be lowered.
If it is less than 00, the effect of improving the stability due to the addition of the PEGylated phospholipid may be reduced.

In the present invention, when the phospholipid having a polyethylene glycol chain is contained in the emulsifier (C), the above-mentioned PEGylated phospholipid is contained in the emulsifier as the component (C) in an amount of 0.1 to 30% by weight. %, Preferably 0.5-20
It may be made to be a weight%. At this time, if the amount used exceeds 30% by weight, the hydrophilicity of the component (C) as a whole becomes high, and as a result, the stability of the finally prepared perfluorocarbon emulsion may decrease.
On the other hand, if it is less than 0.1% by weight, the effect of improving the stability due to the addition of the component may not be sufficiently obtained. The aqueous phase component, which is the component (D) in the present invention, is an aqueous solvent that constitutes the continuous phase of the emulsion and is not particularly limited as long as it is physiologically acceptable, and among those generally used for pharmaceutical applications. Used to choose from. Specific examples of such an aqueous phase component include, for example, distilled water for injection, physiological saline, phosphate buffer, phosphate buffered saline, lactated Ringer's solution, and the like, and if necessary, an electrolyte solution and the like. Can also be used.

Further, the aqueous phase component which is the component (D) of the present invention is, of course, in addition to the above-mentioned aqueous solvent, a tonicity agent such as glycerol, sucrose, glucose, mannitol or polyethylene glycol, It is also possible to use stabilizers such as dextran. In the present invention, the amount of the component (D) used may be adjusted as the balance of the components (A), (B) and (C). Further, in the perfluorocarbon emulsion of the present invention, an antioxidant such as tocopherol may be used in addition to the above components as long as it does not affect the stability of the finally prepared perfluorocarbon emulsion. Next, a method for producing the perfluorocarbon emulsion of the present invention will be described. In the method for producing a perfluorocarbon emulsion of the present invention, for example, an emulsifier (C) such as egg yolk phospholipid is added in advance to distilled water for injection (aqueous phase component D) that has been made isotonic with glycerol or the like, and well dispersed, Specifics of the mixture of the component (C) and the component (D) having the perfluorocarbon compound (A) and a perfluoroalkyl group and an alkyl group in the molecule represented by the general formula (1). And the compound (B) are added and emulsified under the required conditions.

In the production method of the present invention, for example, an emulsifier (C) such as egg yolk phospholipid is previously added to distilled water for injection (aqueous phase component D) that has been made isotonic with glycerol or the like and well dispersed. While circulating the mixture of the component (C) and the component (D) under specific high pressure conditions, the perfluorocarbon compound (A) and the compound represented by the general formula (1) (B) are added to the circulating liquid. ) And are added for preliminary emulsification, and then main emulsification is performed under the required conditions. In particular, in the present invention, when preparing a perfluorocarbon emulsion, a high pressure jet flow emulsification method is used to utilize the shear force generated between liquids of two substantially opposed jet flows under specific conditions. It is possible to prepare a perfluorocarbon emulsion with extremely high stability by emulsifying and then using this preparation method, the particle size becomes extremely fine, so not only heat sterilization but also filter sterilization is possible. It is possible to obtain various emulsions. In the present invention,
In order to finally obtain a fine and stable perfluorocarbon emulsion, it is desirable to carry out preliminary emulsification in advance before emulsification by the above-mentioned high pressure jet flow emulsification method. The emulsifying machine used for the preliminary emulsification and the conditions for the preliminary emulsification are not particularly limited, but it is preferable to use the high-pressure jet flow emulsification method, whereby the continuous operation from the preliminary emulsification to the main emulsification can be performed.

The high-pressure jet flow emulsification method is, as shown in FIG. 1, jetted a raw material component liquid mixture such as an aqueous phase component containing an emulsifier as a jet flow under an ultrahigh pressure of 150 MPa or more. By colliding the jet stream jetted with the wall surface 1 arranged facing each other and reversing the jet stream in a direction substantially opposite to the jet stream direction, the shearing force generated when the jet stream collides, and the two This is a method of performing emulsification by utilizing the shearing force generated between the liquids of the jet streams facing each other (between the flow of the solid arrow in FIG. 1 and the flow of the dashed arrow facing each other) (in the present specification, This method is called the reverse method). Here, "substantially opposite" means a direction in which a shear force can be generated between the jet flow jetted from the jet port and the liquid, and not only the direction parallel and opposite to the jet flow direction. , But also in the opposite direction but having a slight angle with the jet flow direction. Moreover, when performing pre-emulsification, as shown in FIG.
While jetting and circulating as a jet stream under a high pressure of 50 MPa, for example, a component corresponding to an oil phase is injected into the jet stream from the injection tank 4 so that the liquid-liquid between the two jet streams (see FIG. 2). It is also possible to perform the pre-emulsification by utilizing the shearing force generated between the flow indicated by the solid arrow and the flow indicated by the broken arrow (this method is referred to as a dual feed method in the present specification).

That is, the following method is exemplified as a preferred embodiment of the method for producing the perfluorocarbon emulsion of the present invention. An emulsifier (C) such as egg yolk phosphatidylcholine is added in advance to distilled water for injection (aqueous phase component D) isotonic with glycerol or the like and well dispersed. Next, the mixture of the component (C) and the component (D) is
It is jetted as a jet stream under a pressure of 50 to 250 MPa, preferably 70 to 150 MPa and circulated. At this time, cooling is performed using the cooling coil 2 or the like, if necessary. When a phospholipid having a polyethylene glycol chain is used as one of the emulsifiers (C), it may be added to the emulsifier (C) at the above step, but here, another aqueous phase component (here (( Foaming can be suppressed by injecting into the above jet flow a mixed solution in which a phospholipid having a polyethylene glycol chain is dispersed in (D ′)). At this time, the injection of the mixed solution in which the phospholipid having a polyethylene glycol chain is dispersed may be natural injection or forced injection. In this case, the natural injection means to be naturally injected under the reduced pressure generated by the circulating jet flow, and the forced injection is
It refers to forcible pressure injection from the injection port using a syringe or the like.

A mixture of the perfluorocarbon compound (A) and the compound (B) represented by the general formula (1) is injected from the injection tank 4 into the circulating jet stream described above. At this time, the injection of the mixture of the component (A) and the component (B) may be either natural injection or forced injection, as described above. Preliminary emulsification is carried out by utilizing the shearing force generated between the liquid and the liquid to be injected with the circulating jet flow or the liquid (between the flow indicated by the broken arrow and the flow indicated by the solid arrow in FIG. 2) (dual feed). Law). Then, the preliminary emulsion obtained above was treated with 150M.
It is jetted as a jet flow under a pressure of Pa or more, preferably 200 MPa or more, and the jet flow is made to collide with the wall surface 1 arranged facing the injection port to reverse the direction substantially opposite to the direction of the jet flow. Let At this time, if necessary, the back pressure is adjusted to about 10 to 15 MPa. Also,
If necessary, the circulating fluid is cooled by using the cooling coil 2 or the like.

By the above, the jet stream is directed to the wall surface 1
The shearing force generated when the jet collides with the liquid, and the shearing force generated between the liquid and the liquid of the jet stream (between the flow indicated by the solid arrow and the flow indicated by the dashed arrow facing each other in Fig. 1) that are substantially opposite to each other. Then (reverse method), the main emulsification is carried out while circulating until the average particle diameter of the oil phase as the dispersed phase becomes 300 nm or less, preferably 200 nm or less. Incidentally, in the above method, the addition of the tonicity agent is not limited to before emulsification, after preparing the perfluorocarbon emulsion by the method of the present invention, that is, after performing high pressure jet flow emulsification, glucose and electrolyte Finally, isotonicity can also be achieved by adding water for injection (or sterile purified water) in which glycerin or the like is dissolved. By the production method described above, a perfluorocarbon emulsion having excellent stability, which is the object of the present invention, can be prepared.

The production method of the present invention is a commercially available emulsifying apparatus, such as "DeBEE 2000" or "Mini DeBEE", which is an ultrahigh pressure jet flow emulsifying apparatus manufactured by BEE international (Israel).
Alternatively, it can be carried out using an apparatus similar to these.
These emulsifiers are characterized in that they can generate a jet stream under an ultrahigh pressure of 140 MPa or more, and can prepare a very fine emulsion even with a small number of passages as compared with the conventional emulsifiers. Is suitable for.

[0036]

EXAMPLES Next, the perfluorocarbon emulsion of the present invention will be described more specifically by way of examples, but the present invention is not limited to these examples. [Preparation of Perfluorocarbon Emulsion] A perfluorocarbon emulsion was prepared according to the composition and emulsification method shown in Table 1. Examples 1, 5, 6 and Comparative Examples 2-4 Distilled water for injection in which glycerol was previously dissolved (first
In the table, the mixture in which the emulsifier (C component) was added to the D component) and well dispersed was first sufficiently dispersed using a magnetic stirrer. Then, this dispersion was continuously circulated at 100 MPa by a dual feed method using a high pressure jet flow emulsifying machine (DeBEE2000, manufactured by BEE international), while the perfluorocarbon (A
Component) and perfluoroalkyl oleate (component B) or its comparison product (component B ') (comparative example 2)
(A component only) was gradually injected, and preliminarily emulsified by further circulating 5 times at 100 MPa. Then the same high pressure jet flow emulsifier (DeBEE2000, BEE internati
1) at a back pressure of 7 to 12 MPa while cooling the circulation part by a reverse method using
A perfluorocarbon emulsion was prepared by passing the solution 0 times to carry out the main emulsification.

Examples 2, 4, 7 and 8 As in Example 1, distilled water for injection (component D in Table 1) in which glycerol was previously dissolved was added to an emulsifier (component C).
The mixture that was added and was well dispersed was first thoroughly dispersed using a magnetic stirrer. This dispersion was then applied to a high pressure jet emulsifier (DeBEE2000, BEE internati
onal) and the dual feed method
PE as C'component separately while continuously circulating at MPa
G-phosphatidylethanolamine was added to distilled water for injection, and a well-dispersed mixed solution was gradually and naturally injected, and then perfluorocarbon (A component) and perfluoroalkylolate (B component) or a comparative product (B component) was added. The mixture of the'components' was gradually injected, and then preliminarily emulsified by circulating 5 times at 100 MPa. Then same high pressure jet flow emulsifier (DeBEE200
(Manufactured by BEE International Co., Ltd.) was used to carry out main emulsification by cooling the circulation part by a reverse method at 250 to 270 MPa and a back pressure of 7 to 12 MPa for 10 times to prepare a perfluorocarbon emulsion. .

Example 3 An emulsifier (component C) was added to distilled water for injection (component D) in which glycerol had been previously dissolved, and the mixture was well dispersed in warm water. The mixture was mixed with Clairix (Model CLM-0.8) which is a rotary homogenizer. S, manufactured by Createch Co., Ltd., while stirring at 8000 rpm, a mixture of PEGylated phosphatidylethanolamine as a C ′ component was added to distilled water for injection (D component) and well dispersed. A mixture of perfluorocarbon (component A) and perfluoroalkyl oleate (component B) was gradually added in advance, and then preliminarily emulsified by stirring at 10,000 rpm for 5 minutes. After that, the same high-pressure jet flow emulsifying machine (DeBEE2000, manufactured by BEE international) as above
25 while cooling the circulation section by the reverse method.
A perfluorocarbon emulsion was prepared by performing main emulsification by passing 10 times at 0 to 270 MPa and a back pressure of 7 to 12 MPa.

Comparative Example 1 Distilled water for injection in which glycerol was previously dissolved (first
In the table, the mixture in which the emulsifier (C component) was added to the D component) and well dispersed was first stirred at 5000 rpm using a rotary homogenizer CREAMIX (Model CLM-0.8S, manufactured by Createch Co., Ltd.). However, only the perfluorocarbon (A component) was gradually added without mixing the B component, and further preliminarily emulsified by stirring at 10,000 rpm for 5 minutes. Thereafter, a microfluidizer (Model 110Y, manufactured by Microfluidics) was used to carry out main emulsification by passing 10 times at 138 MPa, which is the upper limit of operating pressure, while cooling, to prepare a perfluorocarbon emulsion.

[0040]

[Table 1]

[0041]

[Table 2]

The following tests were carried out and evaluated for each of the above prepared perfluorocarbon emulsions. [Filter Filtration Test] A filter filtration test was performed on each perfluorocarbon emulsion that was left standing at 4 ° C. for 48 hours after being prepared according to the above-mentioned Examples or Comparative Examples in order to attempt sterilization by filtration of each emulsion. That is, 5 ml of each emulsion was taken into a disposable syringe for injection of 10 ml, and a 0.22 μm cartridge filter (ce
llulose acetate) was attached and a filtration test was performed. As a result, all of the emulsions obtained in Examples 1 to 8 could be filtered, but in Comparative Examples 1 to 4, the filter was clogged around 1 to 2 ml, and filtration could not be performed.

[Emulsion Stability] For each perfluorocarbon emulsion prepared in the above Examples or Comparative Examples, each emulsion was placed in a glass vial, sealed and heat sterilized (1
(21 ° C., 20 minutes), samples were taken over time while storing at 4 ° C. and 20 ° C., and the particle size distribution was measured to evaluate the emulsion stability. The particle size distribution is CH
Using DF2000 (MATEC APPLIED SCIENCES), measurement was performed by the capillary hydrodynamic fractionation method, and the volume average value was obtained. The results are shown in Table 2.

[0044]

[Table 3]

As shown in Table 2, as compared with an emulsion prepared by a general method by a conventionally used emulsifying machine,
All the perfluorocarbon emulsions prepared by the method of the present invention using a high pressure jet flow emulsifier had a fine particle size and showed high stability. [Rat Administration Test] In order to confirm the safety of the perfluorocarbon emulsion of the present invention, the emulsion prepared in Example was actually administered to a rat and the condition was observed. For the test, 8-week-old Wister male rats were used, and the right femoral artery of the rat was cannulated under pentobarbital anesthesia, and the emulsion prepared in the example from the femoral vein 30 ml / k.
g was injected with a syringe pump over 20 minutes. The condition of the rat was observed during the administration and for 8 hours after the administration to examine the influence of the administration of the perfluorocarbon emulsion. As a result, no change was observed in the rat's condition in any of the emulsions. It was confirmed that there was no problem with the safety of the perfluorocarbon emulsion.

[0046]

According to the present invention, a perfluorocarbon emulsion having a specific composition is emulsified under a specific condition by using a high pressure jet flow emulsifying machine, whereby the stability is excellent and the particle size is very fine. A perfluorocarbon emulsion that can be sterilized by filtration can be prepared.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing an example of a high pressure jet flow emulsification method (reverse method) according to the present invention.

FIG. 2 is a schematic diagram showing an example of a preliminary emulsification method (dual feed method) of the high-pressure jet flow emulsification method according to the present invention.

[Explanation of symbols]

1: Wall surface (collision surface) 2: Cooling coil 3: Back pressure valve 4: Infusion tank 5: Injection valve

Continued front page    (72) Inventor Yoshikazu Takeuchi             6-15-7 Takaoka, Okubo-cho, Akashi-shi, Hyogo F term (reference) 4C076 AA17 BB12 CC14 DD63 EE51                       FF43                 4C085 HH09 JJ03 KB39 LL01                 4C206 AA01 BA08 MA03 MA05 MA42                       MA86 ZA52

Claims (9)

[Claims] 1. (A) Perfluorocarbon compound 10
~ 80% by weight, (C) 0.1 pharmaceutically acceptable emulsifier
10% by weight, and (D) an aqueous phase component, which is a perfluorocarbon emulsion, further comprising (B) a compound represented by the following general formula (1) in an amount of 0.1 to the above (A) component. A perfluorocarbon emulsion characterized by containing up to 90% by weight. R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. ) 2. A compound (B) represented by the general formula (1).
Is a liquid at a temperature of 40 ° C. The perfluorocarbon emulsion according to claim 1. 3. The perfluorocarbon emulsion according to claim 1, wherein the medically acceptable emulsifier (C) contains a phospholipid. 4. The medically acceptable emulsifier (C) is a phospholipid and a nonionic surfactant in the ratio of 10:90 to 99: 1.
The perfluorocarbon emulsion according to claim 1, which is a mixture (weight ratio). 5. The perfluorocarbon emulsion according to claim 3, wherein the phospholipid constituting the medically acceptable emulsifier (C) contains a phospholipid having a polyethylene glycol chain. 6. A polyethylene glycol chain having a molecular weight of 40 in a phospholipid having a polyethylene glycol chain.
The perfluorocarbon emulsion according to claim 5, which is 0 to 20000. 7. (A) Perfluorocarbon compound 10
-80% by weight, (B) 0.1-9 with respect to the component (A)
0% by weight of a compound represented by the following general formula (1), (C)
A mixture of 0.1 to 10% by weight of a medically acceptable emulsifier and (D) an aqueous phase component is jetted as a jet stream under a pressure of 150 MPa or more, and the mixture is applied to a wall surface facing the jet port. A method for producing a perfluorocarbon emulsion, which comprises emulsifying by colliding jet streams and reversing them in a direction substantially opposite to the direction of the jet streams. R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. ) 8. (A) Perfluorocarbon compound 10
-80% by weight, (B) 0.1-9 with respect to the component (A)
0% by weight of a compound represented by the following general formula (1), (C)
A method for producing a perfluorocarbon emulsion comprising 0.1 to 10% by weight of a medically acceptable emulsifier and (D) an aqueous phase component, which comprises (C) a medically acceptable emulsifier and (D) ) 50-250MP of mixture with water phase ingredients
Pre-emulsification was carried out by jetting as a jet stream under the pressure of a and by injecting a mixture of (A) a perfluorocarbon compound and (B) a compound represented by the general formula (1) into the jet stream. After that, the obtained preliminary emulsion is added to 15
It is jetted as a jet stream under a pressure of 0 MPa or more, and the jet stream is collided with a wall surface facing the jet port, and the jet stream is inverted in a direction substantially opposite to the direction of the jet stream to emulsify. A method for producing a perfluorocarbon emulsion, comprising: R ¹ —X—R ² (1) (In the formula, R ¹ is an alkyl group having 6 to 12 carbon atoms and having a perfluoroalkyl moiety having 6 or more carbon atoms, and R ² is 1 carbon atom.
2 to 22 represents an alkyl group, and X represents Represents However, R ³ is an alkylene group having 2 to 8 carbon atoms. ) 9. A perfluorocarbon emulsion produced by the method according to claim 7 or 8, wherein the average particle size of the dispersed phase is 200 nm or less.