JPH04356417A - Fat emulsion for intravenous injection - Google Patents

Abstract

PURPOSE:To obtain a fat emulsion for intravenous injection, having a fine particle diameter, a small change in the particle diameter of fat particles at high temperatures or in preserving for a long period and high safety, rapidly metabolized, administrable even to patients suffering from hepatopathy, etc., and hardly causing emboli in the reticuloendothelial systems of lung, liver or spleen. CONSTITUTION:A fat emulsion for intravenous injection containing at least one selected from phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, phosphatidylpolyethylene glycol, diphosphatidylethylene glycol and diphosphatidylpolyethylene glycol expressed by formulas I to III [R<1> to R<4> are 6-32C, preferably 12-18C saturated or unsaturated acyl; (m) is a number of 1-10; (n) is an integer of 1-150] as a main emulsifying agent in which >=60% aforementioned emulsifying agent is preferably at least one emulsifying agent selected from the above-mentioned compounds. This fat emulsion for the intravenous injection has the above-mentioned effects.

Description

[Detailed description of the invention]

0001

[Industrial Application Field] The present invention provides an intravenous injection with small particle size and excellent stability that is rapidly metabolized after administration and has a low possibility of causing embolism in the reticuloendothelial system such as the lungs, liver, and spleen. The present invention relates to fat emulsions for use.

0002

BACKGROUND OF THE INVENTION Fat emulsions are used parenterally to provide nutrition to patients as an energy source or essential fatty acid source, and are provided in the form of fine particles by emulsifying fats and oils with an emulsifier. In 1960, Wretlin developed Intralipid [a commercially available fat emulsion for intravenous injection (trade name sold by Otsuka Pharmaceutical Co., Ltd.)]. Since then, fat emulsions have been made from soybean oil or safflower oil, and soybean lecithin or egg yolk lecithin. Emulsified products have been the mainstream, and there have been no major changes over the past 20 years. Currently, MCT fat emulsions and fat emulsions using chemically synthesized structured lipids are being studied.

0003

Problems to be Solved by the Invention In the conventional fat emulsions mentioned above, the particle diameters of the fat particles are irregular, ranging from 0.2 to 1.2 .mu.m, and sometimes larger than 5 .mu.m. For this reason, when such fat emulsions are injected intravenously, microemboli may occur in reticuloendothelial tissues such as the lungs, spleen, and liver Kupffer cells. moreover,
In the case of liver failure, the uptake of exogenous fat emulsion particles is hindered due to slow metabolism in the liver, and in the case of liver cirrhosis, the pores of the liver endothelial cells through which fat particles pass are reduced and narrowed. Therefore, it was not possible to use anything other than endogenous fat. In addition, since conventional fat emulsions have a relatively slow metabolic rate, they tend to increase the neutral fat concentration in liver tissue, which limits their administration to patients with liver disease. Furthermore, conventional fat emulsions have poor stability at high temperatures and during long-term storage, and have the problem of causing aggregation and creaming. Therefore, it has been desired to develop a fat emulsion for intravenous injection that is rapidly metabolized, has small fat particles, and has excellent stability.

0004

[Means for Solving the Problems] Under these circumstances, the present inventors have conducted extensive research to solve the above problems, and have developed a compound in which the hydroxyl group of a specific polyhydric alcohol is replaced with a phosphatidyl group (hereinafter referred to as a phosphatidyl group). The present invention has been completed based on the discovery that an excellent fat emulsion satisfying the above requirements can be obtained by using a polyhydric alcohol (referred to as a polyhydric alcohol) as the main emulsifier.

That is, the present invention is characterized by containing at least one selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidylpolyethylene glycol, and diphosphatidylpolyethylene glycol as a main emulsifier. The present invention provides a fat emulsion for intravenous injection.

Phosphatidylglycerol and phosphatidylpolyglycerol used in the present invention have the following general formula (1), and phosphatidylethylene glycol and phosphatidylpolyethylene glycol have the following general formula (2).
, diphosphatidyl ethylene glycol, and diphosphatidyl polyethylene glycol have a structure represented by the following general formula (3).

[0007]

[Chemical formula 1]

[0008] Phosphatidylglycerol used in the present invention is widely distributed in nature, and is particularly abundant in plants and bacteria, and can be isolated and prepared from these by extraction operations, but it can also be prepared by known methods [ Biochemistry Experiment Course 3
Chemistry of lipids pp. 294-295 (Tokyo Kagaku Doujin) 1
974 et al.], it can also be easily prepared by using lecithin contained in soybeans, egg yolks, etc. as a raw material and allowing phospholipase D to act in the presence of glycerol. Further, these phospholipids after the transfer can be purified by solvent fractionation, high performance liquid chromatography, etc., if necessary. The phosphatidyl polyglycerol, phosphatidyl ethylene glycol, phosphatidyl polyethylene glycol, diphosphatidyl ethylene glycol, and diphosphatidyl polyethylene glycol used in the present invention can also be produced by using lecithin contained in soybeans, egg yolks, etc. as a raw material, for example, using polyglycerin, It can be easily prepared by allowing phospholipase DM to act in the presence of ethylene glycol or polyethylene glycol. The polyglycerin used here includes those with a condensation degree of 2 to 10, and the polyethylene glycol includes diethylene glycol, triethylene glycol, etc., and various polyethylene glycols having an average molecular weight of 200 to 6,000. These emulsifiers can be used alone or in combination of two or more, and the amount used is not particularly limited, but is 0.1 to 5% by weight based on the fat emulsion.
degree is preferred.

As the emulsifier for the fat emulsion of the present invention, basically only the above-mentioned phosphatidylated polyhydric alcohol is used, but other emulsifiers may be blended within a range of less than 40% by weight. For example, when phosphatidylated polyhydric alcohol is produced using lecithin as a raw material, the raw material lecithin may remain in the phosphatidylated polyhydric alcohol produced, but the remaining amount is less than 40% by weight, especially 20% by weight. It is desirable that it be less than If the emulsifier other than the above-mentioned emulsifier is contained in an amount of 40% by weight or more, satisfactory effects cannot be obtained in terms of the particle size, metabolic rate, etc. of the fat emulsion. In addition, in the present invention, the concentration of phosphatidylated polyhydric alcohol is determined according to the standard oil and fat test analysis method (
It was determined by quantitative determination as described in the Japanese Oil Chemists' Association (ed.).

[0010] Furthermore, the lipids used in the present invention include:
Any oil that is liquid at room temperature may be used, and is not particularly limited, but among them, soybean oil, sesame oil, rapeseed oil, cottonseed oil, safflower oil, olive oil, and structured lipids.
etc. are preferred. The amount of lipid used is not particularly limited, but is preferably about 5 to 25% by weight based on the fat emulsion.

The intravenous fat emulsion of the present invention is produced, for example, as follows. That is, first, the phosphatidylated polyhydric alcohol [(1), (2) or (3
) as an emulsifier, coarse emulsification is performed by a liquid crystal dispersion method via an oil droplet dispersed phase in a liquid crystal, and then fine emulsification is performed by a conventional method. To carry out rough emulsification, first, the phosphatidylated polyhydric alcohols (1) and (2)
) or (3) are mixed with about twice the amount of glycerin and water, respectively, to form a surfactant phase. A lamellar liquid crystal phase is formed by adding a lipid such as soybean oil in an amount equal to or twice that of the surfactant phase little by little while stirring. 3 times to 10 times the amount for this liquid crystal phase
Add twice the amount of water little by little while stirring, and mix at 1000 rpm or more, preferably 5000 rpm using an Ultra Homo mixer.
An oil-in-water emulsion is obtained by rough emulsification for 10 minutes or more at rpm or more. In the emulsification, an oil-in-water emulsion with a small particle size can be obtained by emulsifying using a high-pressure homogenizer or an ultrasonic homogenizer. There are no particular restrictions on the conditions for emulsification, but in the case of ultrasonic emulsification, the
It is preferable to emulsify for 0 minutes or more. Among these emulsifying means, a high-pressure homogenizer is preferred for mass production. After adjusting the pH of the obtained emulsion to 7 to 8 with an aqueous alkaline solution, it was successively filtered several times through a membrane filter with a pore size of 1 to 5 μm, and finally once through a membrane filter with a pore size of 0.4 to 0.5 μm. By filtering, it is possible to obtain a fat emulsion having a relatively uniform particle size of 0.3 μm or less. By dispensing this in a nitrogen atmosphere and sterilizing it using an autoclave, a fat emulsion that can be intravenously injected into a living body can be obtained without a large change in particle size before and after sterilization.

0012

[Examples] The present invention will be explained in more detail below with reference to Examples, but the present invention is not limited thereto.

Example 1 Phosphatidylglycerol derived from soybean lecithin as an emulsifier (prepared by allowing phospholipase D to act on soybean lecithin in the presence of glycerol: "Standard Oil and Fat Test and Analysis Method (edited by Japan Oil Chemists Association)" [5. 3.3
Phospholipid composition] Purity 85 mol% by the method described) 1.
2 g was used and mixed with 2.5 g of 98.5% glycerin and 2.5 g of distilled water. 10 g of soybean oil was added little by little while stirring, and then 83.3 g of distilled water was added little by little while stirring, followed by rough emulsification at 9000 rpm for 30 minutes using an Ultra Homo Mixer (manufactured by Tokushu Kika Kogyo Co., Ltd.). Further, this was ultrasonically emulsified at 240 W for 30 minutes while cooling using an ultrasonic homogenizer (manufactured by Branson). 0.
After adjusting the pH to 7.4 with a 1N aqueous sodium hydroxide solution, the mixture was successively filtered through membrane filters with pore sizes of 3 μm, 1.2 μm, and 0.45 μm. After dispensing this in a nitrogen atmosphere, it was heated at 121 °C for 2 hours using an autoclave.
By sterilizing for 0 minutes, a fat emulsion with small particle size and excellent storage stability was obtained.

Example 2 In Example 1, 1.2 g of phosphatidylglycerol
Instead of phosphatidyl polyethylene glycol 400 (polyethylene glycol 40) derived from soybean lecithin
1.2 g of phospholipase D was prepared by the action of phospholipase D on soybean lecithin in the presence of soybean lecithin. A fat emulsion was obtained which was large in size and had excellent storage stability.

Comparative Example 1 In Example 1, 1.2 g of phosphatidylglycerol
Using 1.2 g of purified egg yolk lecithin instead of Example 1
A corresponding fat emulsion was obtained in a similar manner.

Test Example 1 (High Temperature Stability Test) Examples 1 and 2
Table 1 shows the changes in pH and average particle size of each fat emulsion before and after autoclave sterilization (121° C., 20 minutes) at the final stage of the manufacturing process in Comparative Example 1. The pH was measured using a digital pH meter 225 model (manufactured by Iwaki Glass Co., Ltd.), and the average particle size was measured using a Coulter counter (
Manufactured by Coulter Electronics, Coulter Model N
4).

[0017]

[Table 1]

Test Example 2 (Long-term storage stability test) Example 1
Changes in pH and average particle size when stored for long periods at 4°C and 37°C for the fat emulsions obtained in , 2 and Comparative Example 1 and the commercially available fat emulsion for intravenous injection (Intralipid, manufactured by Otsuka Pharmaceutical Co., Ltd.) was tested. Measurements were performed immediately after production and 10, 30, and 60 days after storage. In addition, Example 1
Regarding the fat emulsion obtained in Comparative Example 1, the average particle size, maximum particle size, particle size distribution, zeta potential, and viscosity were also measured, and the results are shown in Table 2. The results of the long-term storage stability test are shown in Table 3. The maximum particle size and particle size distribution were determined using the fat emulsion particle fixation method (Miki, K., Tanimu
ra, H. ;J. Clin. Electron
Microscopy, 12:855-856 (1
It was calculated by scanning electron microscopy observation using 979) ) and computer image processing.

[0019]

[Table 2]

[0020]

[Table 3]

According to Table 2, the average particle diameter of the fat emulsion of the present invention was 190 ± 2.5 nm, which was significantly smaller than 272 ± 10 nm of the fat emulsion using lecithin as an emulsifier (p<0.01). In addition, regarding the particle size distribution, the particles of 0.5 μm or less were 94.1
±2.5%, and 91.0±1.5% for the emulsion of Comparative Example 1, and the proportion of problematic particles of 1 μm or more was also small. Furthermore, the zeta potential of the fat emulsion of the present invention is low;
The viscosity showed a slightly high value.

Furthermore, Table 3 shows that the fat emulsion of the present invention showed little change in particle size even after long-term storage. Also, 37℃
When stored for more than 30 days, most of the particles of the fat emulsion using lecithin as an emulsifier coalesced into giant particles of about 10 μm, and some were destroyed and had rough surfaces, but the fat emulsion of the present invention No giant particles or fracture images that were thought to have coalesced were observed.

Test Example 3 (Stability test under harsh centrifugation conditions)
The fat emulsions obtained in Example 1 and Comparative Example 1 were heated at 37°C for 1
The state of the particles was observed after centrifugation at 572×g for 6 hours. As a result, in the fat emulsion of Comparative Example 1, the fat particles showed a strong tendency to agglomerate, whereas in the fat emulsion of the present invention, the tendency to agglomerate was weak, and almost no particle breakage was observed.

Test Example 4 (Phytokinetics) Using a 3H-labeled emulsifier, a fat emulsion obtained in the same manner as in Example 1 and Comparative Example 1 was administered to rats via bolus vein,
The pharmacokinetics and metabolic rate of fat emulsions were investigated. Figure 1 shows the changes in serum neutral fat concentration after administration of the fat emulsion. As a result, serum neutral fat rapidly increased to 391 ± 53 mg/dl for the fat emulsion of Comparative Example 1 5 minutes after administration, whereas it only slightly increased to 146 ± 12 mg/dl for the fat emulsion of the present invention. (p<0.01). Figure 2 shows changes in neutral fat concentration in liver tissue. As a result, with the fat emulsion of the present invention, the neutral fat in the liver tissue increased to 120% from 5 minutes after administration.
It was found that the increase in triglycerides was small after 30 minutes, and that there was little accumulation of triglycerides in the liver tissue. In addition, the administered 3
As shown in Table 4, the distribution rate of the H-labeled emulsifier to the main tissues was higher for the fat emulsion of the present invention than for the fat emulsion of Comparative Example 1.
After a few minutes, a high distribution rate was already observed in the liver.

[0025]

[Table 4]

[0026] Furthermore, when the metabolic process of fat particles in liver tissue was observed using an electron microscope, it was found that in the fat emulsion of the present invention, fat particles were actively taken up by hepatocytes, and most of the fat particles were metabolized 60 minutes after administration. The metabolic rate in the liver was faster than that of the fat emulsion of Comparative Example 1. In other words, the fat emulsion of the present invention is quickly taken up into the liver after administration, so it disappears quickly from the blood and is also well metabolized in the liver tissue.

[0027]

[Effects of the Invention] As described above, the intravenous fat emulsion of the present invention has a small particle size, and the change in the fat particle size is small even under high temperature or long-term storage, and is an indicator of peroxide production. The degree of pH reduction is also very stable, being equal to or lower than that of conventional fat emulsions using egg yolk lecithin as an emulsifier. Furthermore, the intravenous fat emulsion of the present invention comprises (1
) The particle size is small, making it difficult to cause embolism in the reticuloendothelial system, etc. (2) Metabolism in the blood and liver is fast, so it does not accumulate in the liver and can be administered to patients with liver dysfunction; (3) Compared to conventional fat emulsions using lecithin as the main emulsifier, there is no accumulation of phospholipids themselves, and (4) especially when phosphatidylglycerol is used, the decomposition product is glycerin.
This itself has advantages such as being used as an energy source.

[Brief explanation of drawings]

FIG. 1 shows changes in serum neutral fat concentration after intravenous administration of a fat emulsion.

FIG. 2 shows changes in liver tissue neutral fat concentration after intravenous administration of fat emulsion.

Claims (2)

[Claims] 1. An intravenous injection characterized by containing at least one member selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidylpolyethylene glycol, and diphosphatidylpolyethylene glycol as a main emulsifier. Fat emulsion for use. 2. Claim 1, wherein 60% by weight or more of the emulsifier is at least one selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidylpolyethylene glycol, and diphosphatidylpolyethylene glycol.
Fat emulsion for intravenous injection as described.