WO1992004886A1

WO1992004886A1 - Fat emulsion for intravenous injection

Info

Publication number: WO1992004886A1
Application number: PCT/JP1991/001283
Authority: WO
Inventors: Hiroshi Tanimura; Yoshio Maniwa; Katsunari Takifuji
Original assignee: Meito Sangyo Co., Ltd.
Priority date: 1990-09-26
Filing date: 1991-09-26
Publication date: 1992-04-02

Abstract

A fat emulsion for intravenous injection containing as the main emulsifier at least one compound selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, phosphatidylpolyethylene glycol and diphosphatidylpolyethylene glycol. The diameter of the fat particle is small and little changes even at high temperature or after being stored for long. This emulsion metabolizes so rapidly that there is only a reduced risk of microembolism caused in the lung, spleen, reticuloendothelial tissue, etc., and is so highly safe that it can be administered to the patients with hepatic diseases.

Description

Specification

Fat emulsion for intravenous injection

Technical field

The present invention relates to a lipid emulsion for intravenous injection which is metabolized immediately after administration, has a low possibility of causing emboli in the retinal system such as lung, liver, spleen, etc., has a small particle size, and is excellent in stability. Things.

Background art

Fat emulsions are used parenterally as a source of energy or essential fatty acids to provide nutrition to patients. Oils and fats are emulsified with an emulsifier and provided in the form of fine particles.

Intralipid (a commercially available intravenous fat emulsion (trade name sold by Otsuka Pharmaceutical Co., Ltd.)) developed by Wret liquid in 1960. Since then, fat emulsion has been based on soybean oil or safflower oil. Emulsions with lecithin or egg yolk lecithin dominate and have not changed much in the last 20 years. Currently, MCT fat emulsions and fat emulsions using chemically synthesized structured lipids are being studied.

In the above-mentioned conventional fat emulsion, the particle diameter of the fat particles is irregular, ranging from 0.2 to 1.2 m, and sometimes large, exceeding 5 x m. For this reason, when such a fat emulsion is injected intravenously, microemboli may be caused in tissues of the retina such as lung, spleen, and liver Kupffer cell (Kupffer cell). Furthermore, in the case of liver failure, the metabolism in the liver is slow, and in the case of cirrhosis, the incorporation of exogenous fat emulsion particles is reduced due to the decrease and narrowing of hepatic endothelial cell pores through which fat particles pass. Because of the hindrance, only the endogenous fat could be used.

In addition, conventional fat emulsions have a relatively low metabolic rate, which increases the triglyceride concentration in liver tissue, which limits the administration to patients with liver disease. In addition, conventional fat emulsions have poor stability at high temperatures or during long-term storage, and have the problem of flocculation and creaming.

For this reason, it is used for intravenous injection because of its fast metabolism, small fat particles and excellent stability The development of a fat emulsion has been desired.

Under these circumstances, the present inventors have conducted extensive research to solve the above-described problems, and as a result, have found that a compound in which the hydroxyl group of a specific polyvalent alcohol is substituted with a phosphatidyl group (hereinafter referred to as phosphatidylated polyvalent alcohol). ) Was used as the main emulsifier to find that an excellent fat emulsion satisfying the above requirements was obtained, and completed the present invention.

Disclosure of the invention

That is, the present invention provides, as a main emulsifier, at least one selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidylpolyethylene glycol, and diphosphatidylpolyethylene glycol. Another object of the present invention is to provide a fat emulsion for intravenous injection characterized in that it contains.

BRIEF DESCRIPTION OF THE FIGURES

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

FIG. 2 shows the change in neutral fat concentration in liver tissue after intravenous administration of a fat emulsion.

BEST MODE FOR CARRYING OUT THE INVENTION

The phosphatidyl glycerol and phosphatidyl polyglycerol used in the present invention are represented by the following general formula (1), phosphatidylethylene glycol and phosphatidyl polyethylene glycol are represented by the following general formula (2), and diphosphatidylethylene glycol and diphosphatidyl Polyethylene glycol has a structure represented by the following general formula (3).

R '0CH ₂

R ² QCH 0 OH

I 1! I

CH ₂ -0-P-0- (CH ₂ -CH-CH ₂₀ ) _m -H (1)

Q

CH ₂ -0-P-0- (CH ₂ CH ₂ 0) _n -H (2)

0 one

R'OCHa H ₂ OR ^:

R ² OCH 0 0 CHOR

CH ₂ -0-P-0- (CH ₂ CH ₂ 0) _n -P-0-CH ₂ (3)

0 one D one

(In the formula, R ′, R ² , R ³ and R ⁴ may be the same or different, and each represents a saturated or unsaturated acyl group having 6 to 32 carbon atoms, preferably 12 to 18 carbon atoms, and m represents 1 to A number of 10 and n is an integer of 1 -150.)

The phosphatidylglycerol used in the present invention is widely distributed in nature and is particularly present in a large amount in plants and bacteria, and can be synthesized and prepared therefrom by an extraction operation. Chemistry Experiment Lecture 3 Lipid Chemistry pp. 294-295 (Tokyo Kagaku Dojin) 1974, etc.], and reacting phospholipase D with lecithin contained in soybean, egg yolk, etc. as raw material in the presence of glycerol Can be easily prepared. Further, these phospholipids after the transfer can be purified by solvent fractionation, high performance liquid chromatography, etc., if necessary.

The phosphatidylpolyglycerol, phosphatidylethylene glycol, phosphatidylpolyethylene glycol, diphosphatidylethylene blendol and diphosphatidylpolyethylene glycol used in the present invention are also contained in, for example, soybeans, egg yolks, etc. according to known methods. Phosphoric acid in the presence of polyglycerin, ethylene glycol or polyethylene glycol, starting from lecithin. It can be easily prepared by the action of DM. Examples of the polyglycerin used herein include those having a degree of condensation of 2 to 10, and polyethylene glycol. Examples of the glycol include diethylene glycol, triethylene glycol, 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. The use amount thereof is not particularly limited, but is preferably about 0.1 to 5% by weight based on the fat emulsion.

As the emulsifier for the fat emulsion of the present invention, basically, only the above-mentioned phosphatidylated polyvalent alcohol is used, but other emulsifiers may be blended within a range of less than 40% by weight. For example, when phosphatidylated polyvalent alcohol is produced using lecithin as a raw material, the resulting phosphatidylated polyvalent alcohol may contain lecithin as a raw material, but the remaining amount is less than 40% by weight. It is particularly desirable that the content be less than 20% by weight. When an emulsifier other than the above-mentioned emulsifier is contained in an amount of 40% by weight or more, a satisfactory effect cannot be obtained in terms of the particle size and metabolic rate of the fat emulsion. In the present invention, the concentration of the phosphatidylated polyvalent alcohol is a value quantified according to the description of the standard method for analysis of fats and oils (edited by Japan Oil Chemists' Society).

The lipid used in the present invention may be any liquid at room temperature, and is not particularly limited. Among them, soybean oil, sesame oil, rapeseed oil, cottonseed oil, safflower oil, and olive oil Oil, structured lipid, etc. are preferred. The amount of the 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, coarse emulsification is carried out by a liquid crystal dispersion method using an oil droplet dispersion phase in a liquid crystal, using the phosphatidylated polyvalent alcohol [(1), (2) or (3) as an emulsifier. It is carried out by fine emulsification by the method. To carry out the coarse emulsification, first, the above-mentioned phosphatidylated polyvalent alcohol (1), (2) or (3) is mixed with glycerin and water in an amount of about twice each, and a surfactant is added. Allow a phase to form. A lamellar liquid crystal phase is formed by adding a small amount of a lipid such as soybean oil to the surfactant phase while stirring the same amount or twice as much as the surfactant phase. Add 3 to 10 times the amount of water to this liquid crystal phase while stirring little by little, and use Ultra Homo Mixer on lOOOOrpm 1 Preferably, an oil-in-water emulsion is obtained by coarse emulsification at 5000 rpm or more for 10 minutes or more. In the fine emulsification, an oil-in-water emulsion having a small particle diameter can be obtained by emulsification using a high-pressure homogenizer or an ultrasonic homogenizer. The conditions for milking are not particularly limited, but in the case of ultrasonic emulsification, emulsification at 100 W or more, preferably 200 W or more, preferably for 10 minutes or more is preferable. Among these refining means, in the case of mass production, it is preferable to use a high-pressure homogenizer.

The resulting emulsion can be adjusted to pH with aqueous rukari solution. After adjusting to ~ 8, filtration is continued several times with a membrane filter with a pore size of 1 to 5ίπι, and finally, once with a membrane filter with a pore size of 0.4 to 0.5 / im. A fat emulsion having a particle size of 0.3 / m or less and having a relatively uniform particle size can be obtained. This is dispensed under a nitrogen atmosphere, and then sterilized using an autoclave, thereby obtaining a fat emulsion that can be injected intravenously into a living body without largely changing the particle size before and after sterilization.

Example

Hereinafter, the present invention will be described in more detail 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 fat and oil test analysis method” (edited by Japan Oil Chemists' Society) [5.3.3 Composition of phospholipids] 1.2 g was used and mixed with 2.5 g of 98.5% glycerin and 2.5 g of distilled water. Added with stirring in small portions soybean oil 10 g thereto, followed by distillation ice 83.3 After added with stirring in small portions g, Ultra Rahomomikisa (special unit Kogyo Co., Ltd.) for 30 minutes the crude emulsion at 9000r _P m did. This was further ultrasonically emulsified at 240 W for 30 minutes while cooling using an ultrasonic homogenizer (manufactured by Branson). After adjusting the pH to 7.4 with a 0. IN sodium hydroxide aqueous solution, the solution was continuously filtered through a membrane filter with a pore size of 3 / m, 1.2 uτη, and 0.45 m. This is dispensed under a nitrogen atmosphere, and sterilized in an autoclave at 121 ° C for 20 minutes to obtain a small particle size. A fat emulsion having excellent storage stability was obtained.

Example 2

In Example 1, phosphatidylglycerol was replaced by 1.2 g of phosphatidylglycerol, and phosphatidylpolyethylene glycol 400 derived from soybean lecithin (prepared by allowing phospholipase D to act on soybean lecithin in the presence of polyethyleneglycol 400: the same as in Example 1) A fat emulsion having a small particle size and excellent storage stability was obtained in the same manner as in Example 1 by using 1.2 g of the compound (purity according to analytical method: 83 mol%).

Comparative Example 1

A corresponding fat emulsion was obtained in the same manner as in Example 1, except that 1.2 g of purified egg yolk lecithin was used instead of 1.2 g of phosphatidylglycerol in Example 1.

Test example 1 (high temperature stability test)

Tables 1 and 2 show changes in pH and average particle size of each fat emulsion before and after the autoclave sterilization treatment (121, 20 minutes) in the final stage of the production process in Examples 1 and 2 and Comparative Example 1.

The pH was measured with a digital PH meter 225 (manufactured by Iwaki Glass Co., Ltd.), and the average particle size was measured using a Coulter Counter (manufactured by Coulter Electronics, Coulter Model N4).

table 1

Test Example 2 (Long-term storage stability test)

The fat emulsions obtained in Examples 1 and 2 and Comparative Example 1 and a commercially available intravenous fat emulsion Intralipid (manufactured by Otsuka Pharmaceutical Co., Ltd.) was tested for changes in PH and average particle size during long-term storage at 4: and 37 :. Measurements were made immediately after production and at 10, 30 and 60 days after storage.

The average particle size, maximum particle size, particle size distribution, zeta potential and viscosity of the fat emulsions obtained in Example 1 and Comparative Example 1 were also measured. The results are shown in Table 2. Table 3 shows the results of the long-term storage stability test. The maximum particle size and particle size distribution were determined by scanning electron microscopy using a fat emulsion particle fixation method (Miki, Tanimura, H .; J. Clin. Electron Microscopy, 12: 855-856 (1979)), and computer It was calculated by image processing.

Table 2

mean ± SD * P + 0.01

Table 3

4 t 37 t fat emulsion storage period

PH average particle size (nm) H average particle size (run)

(Immediately) 6.63 190 ± 2.5 6.63 190 ± 2.5 (10 statements) 6.69 219 ± 9.2 5.65 222 ± 2.2 Example 1

6.30 238 ± 3.7 3.48 257 ± 12.1 (60) 6.23 215 ± 5.3 3.31 252 ± 7.8

(Immediately) 6.44 218 ± 11.8 6.44 218 ± 11.8 (10) 6.66 234 ± 11.6 5.44 241 ± 2.9 Example 2

(30) 6.28 265 ± 2.9 3.41 248 ± 3.4 (60) 6.18 233 ± 4.1 3.23 264 ± 3.1

(Immediately) 6.35 272 ± 10.0 6.35 272 ± 10.0 (10) 6.46 314 ± 10.4 5.72 290 ± 5.4 Comparative Example 1

(30) 5.79 357 ± 8.2 3.49 337 ± 27.2 (60) 4.56 318 ± 8.0 3.09 302 ± 8.0

(Immediately) 7.52 231 ± 13.4 7.52 231 ± 13.4 (10) 7.49 265 ± 11.6 4.81 272 ± 2.1

Pid (30) 6.40 330 ± 12.5 3.72 316 ± 11.6

(60) 6.24 295 ± 8.0 3.59 3-1 ± 4.9

According to Table 2, the average particle size of the fat emulsion of the present invention was 190 ± 2.5ππι, which was significantly smaller than that of the fat emulsion using lecithin as an emulsifier, 2 to 2 ± 10 nm ( _P 0.01). . Regarding the particle size distribution, particles having a particle size of 0.5 jum or less were 94.1 ± 2.5% in the fat emulsion of the present invention and 91.0 ± 1.5% in the emulsion of Comparative Example 1, which was a problem. The ratio of particles of 1 im or more was also small. Further, the zeta potential of the fat emulsion of the present invention showed a low value, and the viscosity showed a somewhat high value.

Further, from Table 3, it was found that the fat emulsion of the present invention showed little change in particle size even after long-term storage. In addition, when stored for 1 day at 3 日 for 30 days, fat emulsions using lecithin as an emulsifier coalesce most of the particles into giant particles of about lO ^ in, and some of them were destroyed and the surface became coarse. Although present, the fat emulsion of the present invention did not show any giant particles or a destructive image considered to be fused.

Test example 3 (stability test under severe centrifugal conditions)

The state of the particles when the fat emulsion obtained in Example 1 and Comparative Example 1 was centrifuged at 37: 1572 X g for 6 hours was observed. As a result, the fat emulsion of Comparative Example 1 showed a strong tendency of agglomeration of fat particles, whereas the fat emulsion of the present invention had a weak tendency of agglomeration and almost no particle destruction was observed.

Test Example 4 (Pharmacokinetics)

Using a ³ H-labeled product as an emulsifier, the fat emulsion obtained in the same manner as in Example 1 and Comparative Example 1 was administered to a rat via a bolus vein, and the pharmacokinetics and metabolic rate of the fat emulsion were examined.

FIG. 1 shows changes in serum neutral fat concentration after administration of the fat emulsion. As a result, 5 minutes after administration, the serum neutral fat increased sharply to 391 ± 53 mg / ^ in the fat emulsion of Comparative Example 1, whereas it increased only slightly to 146 ± 12 mgZo¾ in the fat emulsion of the present invention. (P <0.01).

Fig. 2 shows the change in neutral fat concentration in liver tissue. As a result, it was found that, in the lipid emulsion of the present invention, triglyceride in liver tissue showed a small increase in triglyceride from 5 minutes to 120 minutes after administration, and little accumulation of triglyceride in liver tissue. Also, as shown in Table 4, the distribution rate of the administered ³ H-labeled emulsifier to the main tissues was higher in the liver of the fat emulsion of the present invention than in the fat emulsion of Comparative Example 1 5 minutes after administration, as shown in Table 4. .

Table 4 Lung Liver Spleen Kidney Brain Example 1 5 minutes 3.22 ± 0.34 19.8 ± 1.46 0.96 ± 0.04 1.45 ± 0.13 0.25 + 0.017

60 minutes 0.72 ± 0.02 40.4 Sat-1.88 2.57 ± 0.24 0.64 ± 0.02 0.28 ± 0.009

120 minutes 0.53 ± 0.07 39.2 ± 1 45 1.68 ± 0.11 0.62 person 0.03 0.32 ± 0.021 Comparative example 1 5 minutes 0.75 ± 0.20 7.7 + 0.65 1.17 ± 0.17 0.71 ± 0.12 0.19 ± 0.007

60 minutes 0.52 ± 0.05 22.4 ± 1.70 1.70 ± 0.05 0.97 ± 0.06 0.13 ± 0.005

120 min 0.81 ± 0.08 31.7 ± 2.00 1.87 + 0.17 1.55 ± 0.04 0.15 ± 0.003 mean Sat SB

Dosage: Example 1 29; uCi / kg (2.5 ml / kg as fat emulsion)

Comparative Example 1 138 zCi / kg (2.5 ml / kg as fat emulsion)

When the metabolic process of fat particles in the liver tissue was observed by an electron microscope, the fat particles of the present invention 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 in the fat emulsion of Example 1. In other words, the fat emulsion of the present invention is rapidly taken up by the liver after administration, so that it is rapidly eliminated from the blood, and is well metabolized in liver tissue.

Industrial applicability

As described above, the intravenous fat emulsion of the present invention has a small particle size, the change in the particle size of the fat particles is small even under high temperature or long-term storage, and the pH of PH which is an indicator of peroxide production is small. The degree of reduction is very stable, equivalent to or lower than that of a conventional fat emulsion containing egg yolk lecithin as an emulsifier. In addition, the intravenous fat emulsion of the present invention (1) is less likely to cause emboli such as the reticular system due to its fine particle size, and (2) accumulates in the liver because of its rapid metabolism in blood and liver. It can be administered to patients with liver dysfunction without using it. (3) There is no accumulation of phospholipid itself compared with the conventional fat emulsion using lecithin as the main emulsifier. (4) Especially when phosphatidylglycerol is used. The decomposition product is glycerin, which has advantages such as being used as an energy source.

Claims

The scope of the claims

1. At least one selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidyl polyethylene glycol, and diphosphatidyl polyethylene glycol as a main emulsifier. A fat emulsion for intravenous injection, characterized by comprising:

2. 60% by weight or more of the emulsifier is selected from the group consisting of phosphatidylglycerol, phosphatidylpolyglycerol, phosphatidylethylene glycol, diphosphatidylethylene glycol, phosphatidylpolyethylene glycol and diphosphatidylpolyethylene glycol. 2. The fat emulsion for intravenous injection according to claim 1, which is at least one kind.