JPH06172205A - Fat emulsifier containing acreasin compounds - Google Patents

Abstract

PURPOSE:To provide a fat-emulsifying agent high in the therapeutic effect as an agent for preventing and treating Pneumocystis carinii, little in side reaction, capable of being administered in a high content of an active ingredient, and good in stability. CONSTITUTION:The fat-emulsifying agent contains at least one of acreasin compounds of the formula (R1-CO- is long chain saturated or unsaturated fatty acid residue, organic acid residue which may contain a benzene ring, pyridine ring, O, S or N in the molecule; R2 is H, lower alkyl which may be branched, benzyl, amino-lower alkyl whose amino group may be replaced with a mono or di-lower alkyl; R3 is H, CONH2; R4 is H, OH) as an active ingredient, a phospholipid, a liquid triglyceride, and an aqueous medium. The fat-emulsifying agent preferably contains the active ingredient in a concentration of 0.001-50mg/ml, contains the liquid triglyceride, the phospholipid and the aqueous medium in an weight ratio of 1:0.01-1:2-100, and has an average particle diameter of 50-300nm.

Description

Detailed Description of the Invention

[0001]

The present invention relates to the general formula (1)

[0002]

[Chemical 2]

(In the formula, R 1 -CO- represents a long-chain saturated or unsaturated fatty acid residue or an organic acid residue which may contain a benzene ring, a pyridine ring, an oxygen atom, a sulfur atom or a nitrogen atom in the molecule. R2 represents a hydrogen atom, a lower alkyl group which may have a branched chain, an benzyl group or an amino-lower alkyl group in which an amino group may be substituted with a mono-lower alkyl group or a di-lower alkyl group, and R3 represents A hydrogen atom or a --CONH2 group, and R4 represents a hydrogen atom or a hydroxyl group), and contains at least one selected from the group consisting of acreasins represented by the following formula: an active ingredient, a phospholipid, a liquid triglyceride and an aqueous medium. And a lipid emulsion containing acreasins, which is effective, for example, in the prevention and treatment of Pneumocystis carinii pneumonia. And a method for producing the same.

[0004]

[Prior Art] Pneumocyst
is carinii) is a kind of protozoa, though there is debate about its taxonomic position, and one genus and one species is known to date. It is known that this substance can be a pathogen of pneumonia, and low immunity infants due to congenital immunodeficiency or malnutrition, childhood diseases such as acute lymphocytic or myeloid leukemia, autoimmune diseases in the elderly, lung cancer In the case of malignant tumors, especially when a large amount of antitumor agents, steroids, immunosuppressants are used, or when they are combined with AIDS, toxoplasma, cytomegalovirus, actinomycetes, fungi and other infectious diseases, Pneumocystis・ Carini pneumonia often develops, resulting in death due to respiratory failure.

[0005] Currently, as the drugs reported to be effective against Pneumocystis carinii pneumonia, a combination drug (ST mixture) of sulfamethoxazole and trimethoprim which is an antibacterial agent and pentamidine which is an antiprotozoal drug have been reported. However, since sulfa drugs are highly toxic to AIDS patients and pentamidine itself is highly toxic, their use is limited, and accordingly, the therapeutic effect is also limited.

Under such circumstances, there are few side effects,
As a more effective therapeutic drug, the following general formula (1) of the antibiotics acreasin Aα, Aγ, Dα, Dγ etc.

[0007]

[Chemical 3]

(In the formula, R 1 -CO- represents a long-chain saturated or unsaturated fatty acid residue or an organic acid residue which may contain a benzene ring, a pyridine ring, an oxygen atom, a sulfur atom or a nitrogen atom in the molecule. R2 represents a hydrogen atom, a lower alkyl group which may have a branched chain, a benzyl group or an amino-lower alkyl group in which an amino group may be substituted with a mono-lower alkyl group or a di-lower alkyl group, and R3 represents A drug containing an acreasin represented by a hydrogen atom or a -CONH2 group and R4 represents a hydrogen atom or a hydroxyl group as an active ingredient has been proposed, and a dosage form of a high concentration of acreasins has been demanded from the preparation. .

However, the above-mentioned acreasins are extremely insoluble in water and liquid triglycerides, and also insoluble in chloroform and ether. Also, it is soluble in hydrophilic organic solvents such as alcohols such as methanol, ethanol, isopropanol, n-butanol,
These were unsuitable solvents for injectable formulations.
In addition, when using cholic acid or synthetic surfactant as a solubilizer, it is difficult to obtain a formulation that can be maintained at a high concentration.
Furthermore, there was a problem in terms of safety.

On the other hand, it is known to formulate a poorly soluble drug as a fat emulsion, but none of the conventional fat emulsions can maintain a high concentration, and such a poorly soluble drug is a phospholipid, This is a method in which the poorly soluble drug is dissolved in the liquid triglyceride and uniformly dispersed by the constitution of the liquid triglyceride and the aqueous medium, and the poorly soluble drug was required to be dissolved in the liquid triglyceride. However, as described above, the acreasins have the property of being hardly soluble in liquid triglycerides.

[0011]

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and prevents and treats Pneumocystis carinii pneumonia containing an acreasin represented by the general formula (1) as an active ingredient. In the agent,
It is an object of the present invention to provide a stable fat emulsion which is highly safe and easy to use, and which can be administered with a high content of the above-mentioned acreasins.

[0012]

Means for Solving the Problems and Actions That is, the present invention is based on the general formula (1)

[0013]

[Chemical 4]

(Wherein R 1 —CO— represents a long-chain saturated or unsaturated fatty acid residue or an organic acid residue which may contain a benzene ring, a pyridine ring, an oxygen atom, a sulfur atom or a nitrogen atom in the molecule. R2 represents a hydrogen atom, a lower alkyl group which may have a branched chain, a benzyl group or an amino-lower alkyl group in which an amino group may be substituted with a mono-lower alkyl group or a di-lower alkyl group, and R3 represents A hydrogen atom or a --CONH2 group, and R4 represents a hydrogen atom or a hydroxyl group), and contains at least one selected from the group consisting of acreasins represented by the following formula: an active ingredient, a phospholipid, a liquid triglyceride and an aqueous medium. Is a fat emulsion containing acreasins.

In the present invention, since the above-mentioned active ingredient which is extremely insoluble in water or liquid triglyceride can be made into a fat emulsion, the active ingredient acreasins is about 0.001 mg / ml or more in a fat emulsion, and particularly high concentration. It is a fat emulsion that can be contained up to about 50 mg / ml, which is particularly safe from the viewpoint of formulation because it uses an aqueous medium, and has been completed containing a high concentration of the active ingredient. It has become possible to effectively administer the above active ingredient in vivo.

In the present invention, the fat emulsion refers to fine particles of an O / W emulsion obtained by emulsifying liquid triglyceride with phospholipid, and preferably an emulsion having an average particle size of oil droplets of 50 to 300 nm. . The phospholipid in the present invention may be either a naturally occurring phospholipid or a synthetic phospholipid, and examples thereof include purified egg yolk lecithin and purified soybean lecithin. Further, the liquid triglyceride is not particularly limited as long as it is acceptable in the preparation, and examples thereof include soybean oil, sesame oil, synthetic medium-chain fatty acid triglyceride, etc., and particularly purified from the viewpoint of stability of the preparation. Soybean oil and medium chain fatty acid triglycerides are preferred. Further, in addition, palmitic acid, various fatty acids such as stearic acid and oleic acid as an emulsifying aid, and cholesterol or dibutylhydroxytoluene for the purpose of stabilization, antioxidants such as α-tocopherol and its ester may be appropriately added. Good.

Further, the active ingredient of the present invention is a substance represented by the above-mentioned general formula (1), and these are compounds known as antibiotic acreasins (Japanese Patent Publication No. 59-2035).
0-3, Tetrahedron Letters, 4147-4150 (197
6), Helv. Chim. Acta., 62 (4) 1252-267 (1979), JP-A-3-240727, and JP-A-4-99721).

In the above general formula representing the active ingredient, examples of the group R1 include, for example,

[0019]

[Chemical 5]

[0020]

[Chemical 6]

[0021]

[Chemical 7]

[0022]

[Chemical 8]

[0023]

[Chemical 9]

[0024]

[Chemical 10]

[0025]

[Chemical 11]

And the like. Further, examples of the group R2 include, for example, hydrogen atom, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-
Butyl, pentyl, hexyl, 3-methylbutyl, 2-
A linear or branched lower alkyl group having 1 to 6 carbon atoms such as ethylbutyl and 1-ethylbutyl, a benzyl group, 2-
Amino-lower alkyl groups such as aminoethyl, 3-aminopropyl, 4-aminobutyl, 2-aminopropyl, 2-aminobutyl, etc., amino groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, etc. Examples thereof include amino-lower alkyl groups such as 2-aminoethyl and 3-aminopropyl substituted with a mono-lower alkyl or di-lower alkyl group.

The group R3 is a hydrogen atom or --CON.
H2 may be mentioned, and the group R4 may be a hydrogen atom or a hydroxyl group. In the acreasins represented by the general formula (1), R1 --CO-- is a long-chain saturated or unsaturated fatty acid residue, for example, C14-18 (C14-C18), R2 is a hydrogen atom, and R3 is R3. Is preferably a hydrogen atom, and R4 is a hydrogen atom or a hydroxyl group, and an acreasin derivative is preferable. Further, in the acreasin derivative, R1 --CO-- is myristic acid residue (C14), and R4 is a hydroxyl group. Acid residue (C1
6), acreasin Aγ in which R4 is a hydroxyl group, R1
-CO- is a myristic acid residue, R4 is an acreasin Dα represented by a hydrogen atom, R1 -CO- is a palmitic acid residue, and R4 is an acreasin Dγ represented by a hydrogen atom. Stearic acid residue (C18), Echinocandin C in which R4 is a hydrogen atom, R1-CO- is a linoleic acid residue (C18, two double bonds), Echinocandin B in which R4 is a hydroxyl group, etc. Is mentioned.

Further, as the fat emulsion aqueous medium of the present invention,
For example, water, preferably distilled water for injection or an aqueous medium containing about 0.01 to 0.3 M of a polyhydric alcohol as an isotonicity agent such as glycerin, glucose, saccharose, maltose, etc., and liquid triglyceride 1 It is 2 to 100 parts by weight, preferably 8 to 20 parts by weight with respect to parts by weight.

Then, as a method for preparing the fat emulsion of the present invention, for example, first, the above-mentioned active ingredient, phospholipid and liquid triglyceride are sufficiently kneaded, and simply and thoroughly mixed in a mortar. The kneading may be carried out by using a mixer such as a ribbon blender.
Then, as the aqueous medium, 2 to 100 parts by weight, preferably 8 to 20 parts by weight is used with respect to 1 part by weight of liquid triglyceride, and the kneaded product is mixed with the aqueous medium.
For example, in the case of a small scale, it may be sufficiently dispersed in a mortar, and in an appropriate scale, it may be dispersed using a homogenizer or a kneader. Then, this dispersion mixture is further stirred at a high speed, for example, at a speed of 10,000 to 30,000 rpm with a high speed homogenizer for 4 to 1
Stir for about 0 minutes. After that, the stirring mixed solution is emulsified, for example, 60 to 60 at 10 to 50 KHz.
Ultrasonic treatment method that emulsifies with an ultrasonic homogenizer for about 10 seconds or 300 with a microfluidizer
The emulsification may be carried out by a high-pressure emulsification method in which the emulsification is carried out at a pressure of 1600 Kg / cm ² for about 15 to 50 times, and thus the desired acreasin-containing fat emulsion can be produced in a large amount.

The fat emulsion of the present invention thus obtained contains the active ingredient in an amount of 0.001 to 50 mg / ml.
The composition has 0.01 to 1 part by weight of phospholipid and 2 to 100 parts by weight of aqueous medium with respect to 1 part by weight of liquid triglyceride. The obtained fat emulsion is one in which extremely uniform liposome particles are formed. For example, by treating with a 0.22 μm sterilization filter, a sterilized injectable sterile preparation can be easily obtained. It is also possible to use it as an appropriate formulation. Further, the average particle size of the fat emulsion is preferably 50 nm to 300 n as measured by laser scattered light.
m, and a preparation containing the acreasins at a high concentration of 50 mg / ml was obtained, and an effective drug could be provided. The fat emulsion of the present invention may be injected intravenously, intramuscularly, subcutaneously, intradermally, etc., orally, or may be applied to mucous membranes of the oral cavity, nose, lungs, rectum, vagina, etc., and transdermally. But it's okay. Also, the preferred pH of the fat emulsion of the present invention
The range is from 3 to 10, more preferably from 5 to 8, and further in formulation, parabens, preservatives such as phenol, benzalkonium chloride and benzethonium chloride, salts such as sodium chloride and potassium chloride, An isotonicity agent such as sugars such as glucose and sucrose, and polyhydric alcohols such as propylene glycol and glycerin may be used.

[0031]

EXAMPLES Next, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

[0032]

Example 1 Purified egg yolk lecithin (manufactured by Asahi Kasei: Lo
t. 88100261) 5 g, acreasin Aγ 5 g and 10 g soybean oil prescribed by the Japanese Pharmacopoeia were sufficiently kneaded in a mortar for 3 minutes, and an aqueous solution prepared by dissolving 2 g of glycerin in 90 ml of purified water was added and dispersed. Then, this dispersion liquid was roughly emulsified for about 6 minutes at about 20000 rpm with a high-speed homogenizer (Biomixer-BM-1, manufactured by Nippon Seiki Seisakusho Co., Ltd.) to obtain an ultrahigh pressure homogenizer (Microfluidizer-M-11).
OT, Microfluidics Corporation
(manufactured by Ion) and emulsified about 20 times at a pressure of about 840 kg / cm @ 2 to obtain a fat emulsion. Then, sterile filtration was performed using a 0.22 μm membrane filter to obtain a preparation for injection.

The acreasin Aγ content in the prepared liposome preparation was measured by high performance liquid chromatography (HPLC). The average particle size of the liposome preparation was measured by laser scattered light. Prepared liposo
As for the suspension, the acreasin Aγ content is 45.0 mg / m
1 and having an average particle size of 159.8 ± 81 nm, a preparation in which acreasin Aγ was uniformly dissolved was obtained.

[0034]

Example 2 Stability Test 2.5 g of purified egg yolk lecithin and 0.5 g of acreasin Aγ
And 5 g of soybean oil were sufficiently kneaded in a mortar for 3 minutes, and an aqueous solution in which 1 g of glycerin was dissolved in 45 ml of purified water was added and dispersed. Subsequent operations were performed in the same manner as in Example 1 to prepare a sterile-filtered fat emulsion. After preparation, the mixture was stored at 4 ° C. and 25 ° C. for 4 weeks, and the residual rate of acreasin Aγ after 2 weeks and 4 weeks with time (the acreasin Aγ content at the time of preparation was 100.0%) and average particles. The diameter was measured.

The results are shown in Table 1.

[0036]

[Table 1]

As shown in Table 1, the residual rate of acreasin Aγ after 4 weeks was 95% or more at 4 ° C.
It was 90% or more at 5 ° C. Further, the average particle size was stable at 4 ° C. and 25 ° C., maintaining an average particle size of about 145 nm until 4 weeks later. Therefore, upon storage at 4 ° C, a liposome preparation having a fairly long-term stability was obtained.

[0038]

Example 3 2.5 g of purified egg yolk lecithin, 0.5 g of acreasin Aγ and 5 g of soybean oil were sufficiently kneaded in a mortar for 3 minutes, and 2.5 g of glucose was added and dispersed in an aqueous solution of 45 ml of purified water. . Subsequent operations were performed in the same manner as in Example 1 to prepare a sterile-filtered fat emulsion. The average particle size of the obtained fat emulsion was 151.8 ± 75 nm.

[0039]

Example 4 Purified egg yolk lecithin 0.5 g, acreasin Aγ 0.1 g, and medium-chain fatty acid triglyceride (Triester F810; manufactured by Nippon Surfactant Co., Ltd.) 1 g were sufficiently kneaded in a mortar for 3 minutes to give 0.2 g of glycerin. Was dispersed in 10 ml of sterile distilled water by adding an aqueous solution. Then, this dispersion was used for about 2000 with a high-speed homogenizer (Biomixer-BM-1, manufactured by Nippon Seiki Seisakusho).
The mixture was stirred at 0 rpm for about 6 minutes at high speed, and then, with an ultrasonic homogenizer (manufactured by Nippon Seiki Seisakusho, Model U-300) in a stainless steel container containing ice, at about 20 kHz, 20
The emulsion was treated 30 times with a cycle of resting for 10 seconds and emulsifying for 30 seconds to obtain an emulsified liposome suspension.
A 2 nm fat emulsion was obtained.

[0040]

Example 5 Purified egg yolk lecithin 1 g and acreasin Aα
Using 0.2 g, the same operation as in Example 1 was carried out to prepare a sterile filtered fat emulsion.

[0041]

Example 6 Purified egg yolk lecithin 1 g and aclacin Dα
Using 0.2 g, the same operation as in Example 1 was carried out to prepare a sterile filtered fat emulsion.

[0042]

Example 7 Purified egg yolk lecithin 1 g and acreasin Dγ
Using 0.2 g, the same operation as in Example 1 was carried out to prepare a sterile filtered fat emulsion.

[0043]

INDUSTRIAL APPLICABILITY As described above, according to the present invention, a stable fat emulsion can be formulated as an active ingredient having a high content of acreasins which are poorly soluble in water and liquid triglycerides, and as a result, pneumocystis・ It has become possible to provide effective drugs for the prevention and treatment of carini pneumonia.

Claims (10)

[Claims] 1. A compound represented by the general formula (1): (In the formula, R 1 -CO- represents a long-chain saturated or unsaturated fatty acid residue or an organic acid residue which may contain a benzene ring, a pyridine ring, an oxygen atom, a sulfur atom or a nitrogen atom, and R 2 Is a hydrogen atom, a lower alkyl group which may have a branched chain, an amino group in which a benzyl group or an amino group may be substituted with a mono-lower alkyl group or a di-lower alkyl group.
R3 is a hydrogen atom or --CON
H2 group and R4 represents a hydrogen atom or a hydroxyl group), and contains at least one selected from the group consisting of acreasins represented by the formula: active ingredient, phospholipid, liquid triglyceride and aqueous medium. Acrysin-containing fat emulsion. 2. A fat emulsion, wherein the active ingredient is 0.001 to 50 mg / ml in the emulsion, and 0.01 to 1 part by weight of phospholipid is used with respect to 1 part by weight of liquid triglyceride. Is 2 to 100 parts by weight. 3. In the aclacins represented by the general formula (1), R1 --CO-- is a long chain saturated or unsaturated fatty acid residue, R2 is a hydrogen atom, R3 is a hydrogen atom, and R4 is a hydrogen atom or a hydroxyl group. A certain aclacin derivative.
The described fat emulsion. 4. The fat emulsion according to claim 3, wherein R 1 —CO— is myristic acid residue (C 14) and R 4 is acreasin Aα represented by a hydroxyl group. 5. The fat emulsion according to claim 1, wherein R 1 —CO— is a palmitic acid residue (C 16) and R 4 is acreasin Aγ represented by a hydroxyl group. 6. The fat emulsion according to claim 1, wherein R 1 —CO— is myristic acid residue (C 14) and R 4 is acreasin Dα represented by hydrogen atom. 7. The fat emulsion according to claim 1, wherein R1 --CO-- is palmitic acid residue (C16), and R4 is acreasin Dγ represented by a hydrogen atom. 8. The fat emulsion according to claim 1, wherein the phospholipid is purified lecithin. 9. The fat emulsion according to claim 1, wherein the aqueous medium is water or an aqueous solution containing a polyhydric alcohol. 10. The fat emulsion according to claim 1, wherein the liquid triglyceride is soybean oil or medium chain fatty acid triglyceride.