JPH1160491A - Amphotericin b-containing medicine - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an amphotericin B-containing medicine which has reduced toxicity against kidney by having an accumulative property in the liver in a normal dose, is excellent in safety, and is hence useful in preventing/treating fungal infectious diseases in the liver. SOLUTION: The medicine has an accumulative property in the liver in a normal dose of, for example, 1-100 mg or so per one patient per day. It is preferable that the amphotericin B-containing medicine is prepared in a fat emulsion containing, for example, glycerophosphorylcholine, that an average particle size of the fat particle is 0.1-0.2 μm, and that a content of amphotericin B in the fat emulsion is 1-10 mg/ml.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to amphotericin B.
More specifically, the present invention relates to an amphotericin B-containing preparation having enhanced selective accumulation in the liver.

[0002]

2. Description of the Related Art Fat emulsions composed of vegetable oils, phospholipids and water solubilize water-insoluble / water-insoluble drugs and enable parenteral administration, and are also used as drug carriers (drug carriers). I have been. The characteristic of this drug carrier is that it has a specific affinity for vascular endothelial site, reticuloendothelial system, especially for phagocytes (such as macrophages),
The effect of the drug is exhibited by being incorporated into it. That is, it has been proved that the effect is exhibited by the drug being taken up and accumulated in a site where a large number of poor phagocytes such as an inflammatory site and a cancer site exist, and the effect is maintained by the retention.

[0003] Amphotericin B is an antifungal antibiotic having a polyene structure, and is used as a therapeutic agent for mycosis in clinical applications for indications such as Candida hyperplasia in the digestive tract and fungal deep infections such as Aspergillus. Useful.

[0004] On the other hand, amphotericin B is known to have serious side effects such as nephrotoxicity, and its clinical dose is significantly restricted.

When the liver is infected with a fungus, a fungal liver tumor,
Development of a preventive or therapeutic agent for liver fungal infection with reduced nephrotoxicity due to the presence of multiple Candida liver tumors, multiple fungal liver spleen tumors, multiple liver tumors, and multiple fungal liver tumors Have been.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide an amphotericin B-containing preparation which is useful for the prevention or treatment of fungal infections of the liver and which has an enhanced selective accumulation in the liver. Further, it is an object of the present invention to provide an amphotericin B-containing preparation which has reduced nephrotoxicity of amphotericin B and is excellent in safety.

[0007]

Means for Solving the Problems The present inventors have conducted intensive studies in order to achieve the above object, and as a result, by emulsifying amphotericin B in a fat, it has an ability to accumulate in the liver at a normal dose. However, they have found that it is possible to prepare an amphotericin B-containing preparation having substantially no accumulation property in the kidney and having excellent safety and completed the present invention.

[0008] The "normal dose" of amphotericin B of the present invention is defined as a dose at which sufficient pharmacological effects of amphotericin B can be expected from the conventional knowledge and no substantial side effects such as toxicity occur. It means there is.
Specifically, the daily amount per patient is 1 to 100 m
g is exemplified.

[0009] The term "having accumulation in the liver" means that amphotericin B has a high biodistribution in the liver and preferably has a concentration of at least 10 times within 24 hours after administration of amphotericin B as compared with an organ having no accumulation. It means there is.

A specific embodiment of the present invention is a fat emulsion. Such fat emulsions
Contains amphotericin B, oil component, emulsifier and appropriate amount of water.

The oil component used in the present invention generally includes vegetable oil, fish oil, chemically synthesized triglyceride, and the like. Examples of vegetable oils include soybean oil, olive oil, safflower oil, corn oil, sesame oil, cottonseed oil, peanut oil, castor oil, perilla oil, perilla oil, and the like, and preferably soybean oil is used. As the soybean oil, refined soybean oil is preferred, and more preferably refined soybean oil obtained by further refining the purified soybean oil, for example, by a steam distillation method (the purity is exemplified by 99% or more as triglyceride). is there. Examples of chemically synthesized triglycerides include medium-chain fatty acid triglyceride (1CT), structured triglyceride, long-chain fatty acid triglyceride (tripalmitin, trimyristin, etc.), hard fat, and the like.

The amount of the oil component added is preferably 1 to 50% (w / v), more preferably 5 to 30% in the fat emulsion.
(W / v).

The emulsifier used in the present invention is not particularly limited as long as it can be added to pharmaceuticals, and includes known phospholipids such as egg yolk phospholipid, soybean phospholipid, and hydrogenated products thereof. Is a purified phospholipid. Purified phospholipids are mainly phosphatidylcholine,
It is composed of phosphatidylethanolamine and contains other phospholipids such as phosphatidylinositol, phosphatidylserine, sphingomyelin and the like. In addition, those obtained by substantially removing phosphatidylethanolamine from a purified product of naturally occurring phospholipids may be used, and this is obtained by using a phospholipid such as egg yolk and soybean, and performing organic solvent fractionation by a conventional method. Thereafter, it is obtained by purification with an inorganic adsorbent such as silica gel or alumina. The thus obtained phospholipid mainly consists of phosphatidylcholine (JP-A-60-149524). Further, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidic acid, phosphatidylglycerol, sphingomyelin itself can be used. These phospholipids may be used alone or in combination of two or more. The constituent fatty acids of the phospholipids may be either saturated or unsaturated.

The amount of the phospholipid is 1 to 50 parts by weight, preferably 10 to 30 parts by weight, per 100 parts by weight of the oil component. With respect to the whole fat emulsion, it is preferably 1 to 10% (w / v), more preferably 4 to 6% (w / v).
v).

Further, as the emulsifier, a nonionic surfactant can be used in addition to the above phospholipid. Examples of the nonionic surfactant include polyalkylene glycol (for example, polyethylene glycol having an average molecular weight of 1,000 to 10,000, preferably 4,000 to 6,000), and polyoxyalkylene copolymer (for example, average molecular weight 1,000 to 20,000, preferably 2,000
0 to 10,000 polyoxyethylene-polyoxypropylene copolymer), hydrogenated castor oil polyoxyalkylene derivative [for example, hydrogenated castor oil polyoxyethylene-
(20) -ether, same- (40) -ether, same-
(100) -ether etc.), castor oil polyoxyalkylene derivative [eg, castor oil polyoxyethylene-
(20) -ether, same- (40) -ether, same-
(100) -ether, etc.].

The amount of the nonionic surfactant to be added is preferably 5% (w / v) or less, more preferably 1% (w / v) or less, in the fat emulsion.

As described above, the phospholipid and the nonionic surfactant can each be used alone as an emulsifier in the fat emulsion of the present invention. Preferably, phospholipids are used. It is also possible to use both in combination, but in this case, the amount of addition thereof can be appropriately changed so as not to exceed the above-mentioned range, and to be suitable for preparing the preparation of the present invention. .

It is also possible to add other emulsifying auxiliaries known in the art. Examples thereof include linear or branched aliphatic amines having 2 to 22 carbon atoms, such as primary amines and secondary amines, and pharmacologically acceptable salts thereof. Specifically, ethanolamine, propylamine, octylamine, stearylamine, oleylamine, linoleylamine and the like are exemplified as preferable ones. In addition, as these pharmacologically acceptable salts, for example, mineral salts (hydrochloride, hydrobromide, sulfate,
Sulfites, nitrates, phosphates, etc.), organic acid salts (acetate, lactate, succinate, maleate, fumarate,
Malate, tartrate, citrate, methanesulfonate, etc.). The amount of the aliphatic amine added is preferably 1.0% (w / v) in the fat emulsion.
Or less, more preferably 0.5% (w / v) or less.

Further, for example, a fatty acid having 6 to 22 carbon atoms, preferably 12 to 20 carbon atoms, or a pharmaceutically acceptable salt thereof can be added. This fatty acid is not particularly limited as long as it can be added to pharmaceuticals, and may be linear or branched.Specifically, linear stearic acid, oleic acid, linoleic acid, palmitic acid, It is preferable to use linolenic acid, myristic acid and the like. Examples of these salts include pharmacologically acceptable salts such as alkali metal salts (sodium salt, potassium salt, etc.) and alkaline earth metal salts (calcium salt, magnesium salt, etc.). The addition amount of these emulsification aids is preferably 1.0% (w / v) or less, more preferably 0.5% (w / v) or less in the fat emulsion.

Further, in the present invention, if necessary, a stabilizer, a high molecular substance, a tonicity agent and the like can be added.

Examples of the stabilizer include cholesterol, glycerin or its fatty acid monoester (eg, monopalmitin, monostearin, monoolein, monolinolein, etc.), saccharides, eg, monosaccharides (eg, glucose, fructose, etc.), disaccharides (Eg, maltose, sucrose, etc.), sugar alcohols (eg, sorbitol, xylitol, etc.), antioxidants (eg, tocopherols) and the like. Cholesterols and the like are not particularly limited as long as they can be used for medicine. The amount of these stabilizers added is
In the fat emulsion, it is preferably at most 5% (w / v), more preferably at most 1% (w / v).

Examples of the polymer substance include albumin,
Examples include vinyl polymers (eg, polyvinylpyrrolidone, polyvinyl alcohol, etc.), nonionic surfactants, gelatin, hydroxyethyl starch, and the like. Here, as the albumin, a human-derived one is preferable due to the problem of antigenicity. The amount of these high molecular substances to be added is preferably 5% (w / v) or less, more preferably 1% or less, in the fat emulsion.
% (W / v) or less.

Examples of the tonicity agent include glycerin, monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), sugar alcohols (eg, sorbitol, xylitol, etc.), electrolytes (eg, Sodium chloride, etc.) are exemplified, and it is only necessary that the minimum amount required for isotonicity be added. Moreover, you may add a pH adjuster as needed. Examples of the pH adjuster include sodium hydroxide, hydrochloric acid, various organic acids (eg, acetic acid, citric acid, phosphoric acid, lactic acid, etc.), various amines (eg, Tris, histidine, etc.), various buffers (eg, acetic acid,
Phosphate buffer).

Amphotericin B used in the present invention is a known substance, and its production method is also described in Antibiot. Ann., (1955-195).
6), p579, Chem. Commun., (1986), p413, J. Am. Chem. S
oc., vol. 109, p2821 (1987).

The content of the amphotericin B in the fat emulsion can be appropriately increased or decreased depending on the form and use of the emulsion. Specifically, about 1 to 10 mg / ml is exemplified.

The fat emulsion according to the present invention is produced by a known method, for example, by the following method. That is, a predetermined amount of an oil component (for example, soybean oil), phospholipid, amphotericin B and other additives as required are mixed, and if necessary, heated, and a conventional homogenizer (for example, a high-pressure injection homogenizer) is used. , An ultrasonic homogenizer, etc.) to prepare a water-in-oil type dispersion, then add a necessary amount of water to the dispersion, homogenize again with the homogenizer, and convert to an oil-in-water emulsion. It can be manufactured by the following. Depending on production convenience, additives such as a stabilizer and a tonicity agent may be added after the formation of the fat emulsion (JP-A-56-167161 and JP-A-58-22220). Alternatively, a fat emulsion can be prepared by mixing all the components at the same time and emulsifying them at once. The fat emulsion of the present invention thus prepared is extremely fine, and the average particle diameter of the fat particles is about 0.0
5 to 0.5 μm, preferably 0.1 to 0.2 μm
It is about.

The thus-prepared amphotericin B-containing fat emulsion of the present invention has selective accumulation in the liver at normal doses compared to conventional amphotericin B preparations, but substantially no renal accumulation in the kidney. Does not have integration. Therefore, the fat emulsion of the present invention is a formulation with reduced toxicity in the kidney and excellent safety, and fungal infections in the liver, such as fungal liver tumors, multiple Candida liver tumors, and multiple fungal liver spleen. It is useful for prevention or treatment of tumors, multiple liver tumors, multiple fungal liver tumors, and the like.

The preparation containing amphotericin B of the present invention has low toxicity, and can be used for humans, dogs, cows, horses, rabbits, mice,
It is useful as a therapeutic agent for mammals such as rats.

The amphotericin B-containing fat emulsion according to the present invention is preferably administered parenterally, such as by injection or continuous infusion, and is administered intravenously, intraarterially, subcutaneously, intradermally, intramuscularly, or topically. In particular, intravenous administration is preferred. The usage / dose is, for example, about 1 to 3 times a day, and the dose is about 1 to 100 mg. The dose can be increased or decreased at any time depending on symptoms, sex, age, and the like.

[0030]

EXAMPLES The present invention will be described in detail below with reference to examples and experimental examples, but the present invention is not limited to these examples.

Example 1 200 g of refined soybean oil as an oil component, 2.5 g of amphotericin B as a drug, 50 g of egg yolk phospholipid as an emulsifier, and an appropriate amount of distilled water for injection were used. Was. Further, this was emulsified under high pressure using a Manton-Gaurin homogenizer. The pH was adjusted to 7-8. As a result, a homogenized fat emulsion containing very fine amphotericin B was obtained. The average particle size of the emulsion was 0.1 to 0.2 μm.

Example 2 A fat emulsion was obtained in the same manner as in Example 1 except that the amount of amphotericin B was changed to 8 g.

Example 3 A fat emulsion was obtained in the same manner as in Example 1 except that egg yolk phosphatidylcholine was used instead of egg yolk phospholipid.

Example 4 A fat emulsion was obtained in the same manner as in Example 1 except that DPPC (dipalmitoylphosphatidylcholine) was used instead of egg yolk phospholipid.

Example 5 A fat emulsion was obtained in the same manner as in Example 1 except that purified sesame oil was used instead of purified soybean oil.

Example 6 Except for using refined olive oil instead of refined soybean oil,
A fat emulsion was obtained in the same manner as in Example 1.

Example 7 A fat emulsion was obtained in the same manner as in Example 1, except that 12 g was used instead of 50 g of yolk phospholipid.

Example 8 A fat emulsion was obtained in the same manner as in Example 1 except that glycerin was added so as to be 22 g per 1,000 ml of the fat emulsion.

Experimental Example 1: Stability 1) Preparing the lipid emulsion of the present invention (Example 1) and visually observing it, no precipitation of amphotericin B was observed. The same was true after refrigerated storage for 10 days. 2) The particle size of the fat emulsion of the present invention (Example 1) was measured by a light scattering method. Immediately after preparation, the average is 0.18 μm, 10
It was almost the same after refrigerated storage for one day.

Experimental Example 2: Toxicity The fat emulsion of the present invention (Example 1) was administered to BALB / c mice (5 weeks old, weighing 23 g on average) at a dose of 4 to 25 mg / day.
It was administered into the tail vein with a body weight of kg and the presence or absence of survival for 8 days was confirmed. As a result, in all of the groups to which the fat emulsion of the present invention was administered, no dying mice were observed, and the survival rate was 10%.
It was 0%. During this time, the weight of the mice increased steadily.

Experimental Example 3: Accumulation in liver The fat emulsion of the present invention (Example 1) was administered to BALB / c mice at a dose of 1 mg / kg body weight in the tail vein, and the distribution over time to each tissue. It was confirmed. Table 1 shows the results.

[0042]

[Table 1]

[0043]

As is apparent from the above description, according to the present invention, there is provided an amphotericin B-containing preparation having an enhanced selective accumulation in the liver, and thus the nephrotoxicity of amphotericin B is reduced. To provide an amphotericin B-containing preparation which is more excellent in safety. It is also expected that this formulation could provide a new clinical application method of amphotericin B.

 ────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Takashi Imagawa 2-25-1, Shodai Otani, Hirakata City, Osaka Prefecture Inside the Midori Cross Research Institute, Inc.

Claims (4)

[Claims] 1. An amphotericin B-containing preparation having an accumulation property in the liver at a normal dose. 2. The preparation according to claim 1, which is in the form of a fat emulsion. 3. The preparation according to claim 2, wherein the particle size is 0.1 to 0.2 μm. 4. The preparation according to claim 1, which is useful for preventing or treating a fungal infection in the liver.