JPH0482824A - Liposome and oil-in-water type emulsion - Google Patents

Abstract

PURPOSE:To obtain oil-in-water type emulsion useful for fields of drug or cosmetic, etc., having liposome of uniform particle diameter and stability in storing for more than two years containing dibasic acid-bonded glycolipid. CONSTITUTION:100mol phospholipid is mixed with 0.1-20mol cholesterol and 0.1-20mol dibasic acid-bonded glycolipid composed of monosaccharide or oligosaccharide containing 2-5 pieces of constituting monosaccharide having ester bonding with 2-7C dibasic acid in an amount of 1-2mol to 1mol monosaccharide and 6-24C fatty acid in an amount of 1-2mol to 1mol monosaccharide and processed in a normal method to obtain liposome having <=0.5mum particle diameter. Oil-in-water type emulsion is obtained by mixing 100 pts.wt. triglyceride and diglyceride with 2-100 pts.wt. dibasic acid-bonded glycolipid having the same composition as the above-mentioned glycolipid. One or both of hydrophilic drug and lipophilic drug are able to be mixed with the liposome and the oil-in-water type emulsion.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to liposomes and oil-in-water emulsions. Liposomes and oil-in-water emulsions are widely used in many fields such as pharmaceuticals and cosmetics.

(Prior Art) In recent years, attempts have been made to apply liposomes and oil-in-water emulsions to pharmaceuticals, cosmetics, and the like. Liposomes are promising for encapsulating fat-soluble drugs and water-soluble drugs to control the sustained release of drugs and as target-directed (delivery system) preparations. Generally, liposomes for pharmaceutical use are required to have a uniform particle size of 0.5ρ or less. In addition, oil-in-water emulsions are promising as target-directed (drug delivery system) formulations for fat-soluble drugs. Oil-in-water emulsions for pharmaceuticals and cosmetics are desired to have storage stability for two years or more.

Conventionally, as liposomes for pharmaceuticals, a method of using ubidecanone-containing liposomes whose membrane surface is coated with a sialic acid-modified natural polysaccharide derivative for target-directed preparations (Japanese Patent Laid-Open No. 6
3-313724) and a method of preparing a freeze-dried preparation from α,α-trehalose mycolic acid ester-containing liposomes (Japanese Patent Application Laid-Open No. 75421/1982). In addition, a method of using an aqueous fat emulsion containing ubidecanone in orally administered nutritional supplements (Japanese Unexamined Patent Application Publication No. 1988-19515) and a method of using an emulsion containing a hydrophilic surfactant and polyhydric alcohol in cosmetics (Japanese Patent Application Laid-Open No. 1987-19515) Kaihei 1-288
Publication No. 330).

(Problem to be solved by the invention) However, the liposome has a uniform particle size of 0.5p.
It is difficult to produce the following, and the emulsion has poor long-term storage stability of 2 years or more, making it difficult to put it into practical use.

An object of the present invention is to provide liposomes with uniform particle size and oil-in-water emulsions that are storage stable for more than 2 years.

(Means for Solving the Problems) The present invention provides: ■ per 100 moles of phospholipids ■ 0.1 to 20 moles of cholesterol; and ■ monosaccharides or oligosaccharides having 2 to 5 constituent monosaccharides per mole of ,
1 to 2 mol of dibasic acid having 2 to 7 carbon atoms and 6 to 2 carbon atoms
This liposome is obtained from a lipid composed of 0.1 to 20 moles of a dibasic acid-bonded glycolipid in which 1 to 2 moles of the fatty acid No. 4 is ester-bonded.

Further, the second invention provides one or two types of triglycerides and diglycerides.
This is an oil-in-water emulsion obtained using a lipid consisting of 2 to 100 parts by weight of the dibasic acid-bonded glycolipid per 100 parts by weight of a mixture consisting of at least one species.

Phospholipids that can be used in the liposomes of the present invention include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, phosphatidylinositol, phosphatidylglycerol, sphingomyelin, egg yolk lecithin, soybean lecithin, and those derived from other animal and plant tissues. , or those obtained by synthesis. These may be used alone or in combination of two or more.

The saccharide that is a component of the dibasic acid-bound glycolipid that can be used in the liposomes and oil-in-water emulsions of the present invention is preferably a monosaccharide or an oligosaccharide having 2 to 5 constituent monosaccharides.

If the number of constituent monosaccharides is 6 or more, production is difficult and undesirable.

Examples of monosaccharides include pentose sugars such as arabinose, xylose, lyxose, ribose, and ribulose sugar, hexose sugars such as galactose, glucose, sucrose, mannose, sorbose, tagalose, psicose, fructose, etc.
monodeoxy sugars deoxyribose, isolobose,
Deoxyhexose, deoxyaldose, deoxyclose, deoxygulose, deoxyclose, fucose, rhamnose, deoxyarabinohexose, deoxyxylohexose, dideoxy sugar digitoxose,
Examples of oligosaccharides include dideoxylyxohexose, voivinose, arabinose, abecose, collidose, tiberose, and paradose. Examples of oligosaccharides include trehalose,
Examples include maltose, cellbiose, sucrose, lactose, etc. in which 2 to 5 of the above monosaccharides are bonded.

The dibasic acid that is a component of the dibasic acid-bound glycolipid that can be used in the present invention preferably has 2 to 7 carbon atoms. If the number of carbon atoms is 8 or more, it is difficult to manufacture and is not preferred. Examples of dibasic acids having 2 to 7 carbon atoms include malonic acid, succinic acid, glutaric acid, adipic acid, and pimelic acid.

The fatty acid constituting the dibasic acid-bonded glycolipid that can be used in the present invention preferably has 6 to 24 carbon atoms. If the number of carbon atoms is 5 or less, liposomes and emulsions will not be formed, and if the number of carbon atoms is 25 or more, production will be difficult, which is not preferred. Fatty acids having 6 to 24 carbon atoms include caproic acid, enanthic acid, caprylic acid, undecanoic acid, lauric acid, tridecanoic acid, myristic acid, pentadecanoic acid, valmitic acid, palmitooleic acid, heptadecanoic acid, stearic acid, and oleic acid. , elaidic acid, linoleic acid,
Examples include lysyllic acid, stearolic acid, lysyllic acid, arachidic acid, arachidonic acid, hehenic acid, eicosabencitric acid, docosahexaenoic acid, erucic acid, brassic acid, lignoceric acid, and the like.

Furthermore, the present invention uses Rhodococcus erythropolis as a dibasic acid-binding glycolipid.
The succinyl trehalose lipid produced by th7□ can be used. Succinyl-trehalose lipids that can be used in the liposomes and oil-in-water emulsions of the present invention include disuccinyl divalmitoyl trehalose, disuccinyl distearyl trehalose, disuccinyl dioleoyl trehalose, succinyl dipalmitoyl trehalose, succinyl distearyl trehalose, succinyl Examples include valmitoyltrehalose, succinylstearyltrehalose, succinyldioleoyltrehalose, and succinyloleoyltrehalose.

The succinyl trehalose lipid is disclosed in JP-A-62-8
It is manufactured by the method disclosed in Japanese Patent No. 3896. This method uses strain 5D-74 belonging to Rhodococcus erythropolis, which was isolated as a hydrocarbon-assimilating bacterium, to produce normal alkanes with 6 or more carbon atoms, normal alkenes, long-chain saturated and unsaturated fatty acids, fatty acid esters, and higher alcohols. ,
and aerobically cultured in a medium containing one or more substances selected from oils and fats to produce and collect a succinyl-trehalose lipid mixture.

The liposome of the present invention preferably contains 0.1 to 20 moles of cholesterol and 0.1 to 20 moles of dibasic acid-bound glycolipid per 100 moles of phospholipid. If the cholesterol is less than 0.1 mole per 100 moles of phospholipid, the particle size of the liposome becomes large, and even if it exceeds 20 moles, the effect remains the same. Furthermore, if the dibasic acid-bound glycolipid is less than 0.1 mole per 100 moles of phospholipid, the particle size of the liposome becomes large, and even if it exceeds 20 moles, the effect remains unchanged.

Liposomes can be produced by conventional methods.

For example, phospholipids, cholesterol, and dibasic acid-bound lipids are placed in an eggplant-shaped flask, a small amount of organic solvent is added to dissolve them, the solvent is distilled off to form a thin film, and then water or various filling liquids are added. , can be prepared by applying ultrasound. The organic solvent used is not particularly limited as long as it dissolves phospholipids, cholesterol, and dibasic acid-bound glycolipids, but chloroform, methanol, ethanol, benzene, toluene, ethyl ether, and mixed solvents thereof are used. be able to.

As the encapsulating liquid, in addition to pure water such as distilled water for injection and sterilized water, glucose solutions, physiological saline, buffer solutions, aqueous solutions of various salts, aqueous saccharides, and aqueous solutions of amino acids can be used.

The oil-in-water emulsion of the present invention is prepared by adding water or an aqueous solution to a mixture of 100 parts by weight of a mixture of one or more of triglycerides and diglycerides and 2 to 100 parts by weight of dibasic acid-bound glycolipids. It can be formed by subjecting it to a bath-type sonic bath at ℃. If less than 2 parts by weight of dibasic acid-bound glycolipid is used for 100 parts by weight of a mixture of one type or two or more of triglycerides and diglycerides, an emulsion will not be formed;
Stability does not particularly improve even if the weight part is exceeded.

Triglycerides include soybean oil, rapeseed oil, coconut oil, palm oil, olive oil, safflower oil, cottonseed oil, corn oil,
Oils and fats such as beef tallow, lard, and fish oil, and medium-chain fatty acid triglycerides, diglycerides include soybean oil, rapeseed oil, coconut oil, palm oil, olive oil, safflower oil, cottonseed oil,
Examples include diglycerides and medium-chain fatty acid diglycerides made from corn oil, beef tallow, lard, fish oil, and the like.

As the water, pure water such as distilled water for injection or sterile water may be used, and as the aqueous solution, glucose solution or physiological saline may be used.

As the drug to be encapsulated or mixed in the liposomes and oil-in-water emulsions of the present invention, either a hydrophilic drug or a lipophilic drug, or a mixture of both can be used. Examples of hydrophilic drugs include various anti-inflammatory drugs, lymphokines, anticancer drugs, immunostimulants, physiologically active peptide drugs, antibiotics, antivirals, antiprotozoal drugs, enzyme drugs,
Examples include anti-allergic agents. These can be used by dissolving them in the above-mentioned water or aqueous solution. Examples of lipophilic drugs include anticancer drugs such as anzamitocin, immunostimulants such as TMD-66 and MTI''PE, and phospholipid derivatives.

(Effects of the Invention) The liposomes of the present invention have a particle size of 0.5ρ or less and a small particle size distribution, and are useful as pharmaceuticals for drug delivery systems and cosmetics encapsulating drugs.

Furthermore, the oil-in-water emulsion of the present invention is stable for two years or more after preparation, and is useful as pharmaceuticals and cosmetics containing lipophilic drugs.

(Example) Example 1 Microbial-derived succinyl-trehalose lipid-containing liposome 200 μmoles of egg yolk phosphatidylcholine, 10 μmoles of cholesterol, 10 μmoles of dipalmitoyl disuccinyl trehalose lipid (2 molecules of succinic acid and 2 molecules of palmitic acid bound to trehalose) in 50 m! It was placed in an eggplant-shaped flask. In 10mβ chloroform?
The mixture was dissolved and spread into a thin film using a rotary evaporator while removing chloroborum under reduced pressure. The pressure was reduced using a vacuum pump for 1 hour to remove chloroform. 2 m of physiological saline
1 and shaken vigorously with a portex mixer to peel off the thin film from the walls. Then, it was subjected to an ultrasonic crusher. The diameter of the liposomes was measured using a particle size analyzer. Table 1 shows the average particle size and standard deviation.

Example 2 Glutaryl-glucose lipid-containing liposome Same as Example 1 except that dibalmitoyl diglucryl glucose (two molecules of glutaric acid and two molecules of balmitic acid are bound to glucose) was used instead of the succinyl-trehalose lipid in Example 1. Liposomes were formed accordingly. Table 1 shows the average particle size and standard deviation.

Comparative Example 1 Liposomes were formed from egg yolk phosphatidylcholine and cholesterol according to Example 1 except that no glycolipids were added. Table 1 shows the average particle size and standard deviation.

Table 1 shows that the liposomes containing the glycolipids of the present invention have small particle sizes, and the standard deviation results indicate that they are uniform liposomes with uniform particles.

Example 3 Oil-in-water emulsion containing microorganism-derived succinyl-trehalose lipid medium-chain fatty acid triglyceride ester (MCT: manufactured by NOF Corporation, trade name Panacet 810) and dipalmitoyldisuccinyl-trehalose lipid (trehalose, 2 molecules of succinic acid and 2 molecules of palmitic acid) is combined) and water as shown in Table 2.
They were mixed in different ratios and placed in a bath-type ultrasonic cleaner at 60°C for 5 minutes to create an emulsion. Each was stored at 4°C for 24 months. The emulsified state was observed immediately after preparation, 6 months later, and 24 months later. The results are shown in Table 2.

Example 4 Oil-in-water emulsion containing microorganism-derived succinyl-trehalose lipid Two types of emulsions shown in Table 2 were prepared according to Example 3, except that soybean oil was used instead of medium-chain fatty acid triglyceride ester. Each was stored at 4°C for 24 months, and the emulsified state was observed immediately after preparation, 6 months later, and 24 months later. The results are shown in Table 2.

Example 5 Oil-in-water emulsion containing glutaryl-glucose lipid Example 3 except that the dipalmitoyl disuccinyl trehalose lipid was replaced with dibalmitoyl diglutaryl glucose (two molecules of glutaric acid and two molecules of balmitic acid bound to glucose). Two types of emulsions shown in Table 2 were prepared according to the above. Each was stored at 4°C for 24 months, and the emulsified state was observed immediately after preparation, 6 months later, and 24 months later. The results are shown in Table 2.

Example 6 Oil-in-water emulsion containing glutaryl/glucose lipid Same as Example 4 except that dipalmitoyl disuccinyl trehalose lipid was replaced with dibalmitoyl diglutaryl glucose (two molecules of glutaric acid and two molecules of palmitic acid bound to glucose). Two types of emulsions shown in Table 2 were prepared accordingly. Each was stored at 4°C for 24 months, and the emulsified state was observed immediately after preparation, 6 months later, and 24 months later. The results are shown in Table 2.

Comparative Example 2 Medium-chain fatty acid triglyceride ester (MCT: Nippon Oil & Fats Co., Ltd., trade name Banasate 810) and water were mixed in two proportions shown in Table 3 without adding a monobasic acid-bound glycolipid,
Although each sample was placed in a lotus-shaped ultrasonic cleaner for 5 minutes at 60°C, no emulsion could be created.

Comparative Example 3.4 Table 3 was carried out according to Examples 3 and 4, except that the dipalmitoyl disuccinyl trehalose lipid was replaced with disuccinyl trehalose (two molecules of succinic acid bound to trehalose).
I tried to create each of the two types of emulsions shown below, but I could not create any emulsions.

Comparative Example 5.6 Examples 3 and 4 except that the dibalmitoyl disuccinyl trehalose lipid was replaced with a dipalmitoyl trehalose lipid (two molecules of palmitic acid bound to 1-levalose).
Two types of emulsions shown in Table 2 were prepared according to the method. Each was stored at 4°C for 24 months, and the emulsified state was observed immediately after preparation, 6 months later, and 24 months later. Table 3 shows the results.
It was shown to.

Comparative Example 7.8 An emulsion was prepared according to Examples 3 and 4, except that the dipalmitoyl disuccinyl trehalose lipid was replaced with a dipropionyl disuccinyl trehalose lipid (two molecules of palmitic acid and two molecules of succinic acid bound to trehalose). Two types of each shown in Table 2 were prepared. Each was stored at 4°C for 24 months, and the emulsified state was observed immediately after preparation, 6 months later, and 24 months later. The results are shown in Table 3.

Claims (2)

[Claims] (1) [1] 0.1 to 20 moles of [2] cholesterol per 100 moles of phospholipid, and [3]
1 to 2 moles of dibasic acid having 2 to 7 carbon atoms and 1 to 2 moles of fatty acid having 6 to 24 carbon atoms are ester-bonded to a monosaccharide or an oligosaccharide having 2 to 5 constituent monosaccharides per mole of monosaccharide. A liposome obtained from a lipid consisting of 0.1 to 20 mol of a dibasic acid-bound glycolipid. (2) Per 100 parts by weight of a mixture of one or more triglycerides and diglycerides, a monosaccharide or an oligosaccharide having 2 to 5 constituent monosaccharides, and a dibase having 2 to 7 carbon atoms per mole of monosaccharide. An oil-in-water emulsion obtained using a lipid consisting of 2 to 100 parts by weight of a dibasic acid-bonded glycolipid in which 1 to 2 moles of an acid and 1 to 2 moles of a fatty acid having 6 to 24 carbon atoms are ester-bonded.