WO1991014423A1

WO1991014423A1 - Liposome preparation

Info

Publication number: WO1991014423A1
Application number: PCT/JP1991/000373
Authority: WO
Inventors: Masashi Mukai; Yoshihiro Ishizue; Shinichi Ishikawa; Yuzo Kimura
Original assignee: Otsuka Pharmaceutical Co., Ltd.
Priority date: 1990-03-20
Filing date: 1991-03-20
Publication date: 1991-10-03
Also published as: AU7496991A

Abstract

A liposome preparation comprising a basic drug incorporated in a liposome having a membrane constituent comprising an acidic phospholipid or a mixture of neutral and acidic phospholipids, which can trap the basic drug stably in a sufficient amount.

Description

Ministry

Ribosome preparation

Industrial applications

The present invention relates to a ribosome preparation, more particularly, a liposome containing an acidic phospholipid as a membrane component or a neutral phospholipid and an acidic phospholipid as a membrane component while retaining a basic drug. Liposome preparations.

Conventional technology

More than 20 years have passed since the introduction of ribosomes by Bangham. During this time, ribosomes, which are closed vesicles that have a bilayer composed mainly of lipids and have an aqueous layer inside, are widely used as biomembrane model substances for research on their physicochemical properties. Has been used. On the other hand, liposomes can hold a variety of drugs in the inner water layer or membrane, so they are used as a drag carrier, and ribosomes containing the drug are administered intravenously and the drug is confined to a limited site. There have been many applied researches such as drug delivery systems (DDS), which can be used to get the drug overnight.

However, the drug holding capacity of ribosomes was small and limited, and it was difficult to hold large amounts of drugs. In addition, the structure of the ribosome is stable In many cases, vesicles are destroyed due to aggregation and bonding between liposome particles, and the retained drug leaks to the outer layer, and further, precipitates are formed, and insolubles are precipitated. Is known to occur relatively quickly.

Therefore, despite the fact that many researchers are conducting research on the application of ribosomes to clinical practice, one of the major factors that has yet to commercialize large-scale liposome preparations is the limitation of drug retention capacity. The other is the instability of ribosomes as pharmaceuticals.

Conventionally, the present inventors have also attempted to prepare liposomes holding various basic drugs using various phosphatidylcholines, but have been able to obtain liposomes capable of stably maintaining a sufficient trapping rate. Did not.

The present inventors have made intensive studies to solve the above problems, and have completed the present invention.

Disclosure of the invention

The present invention relates to a liposome preparation containing a basic drug in a ribosome containing an acidic phospholipid as a membrane component or a neutral phospholipid and an acidic phospholipid as a membrane component.

The acidic phospholipid in the present invention is defined more narrowly than ordinary acidic phospholipids, and more specifically, dilauroylphosphatidylglycerol (DLPG) dimyristyl. Phosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol (DPPG), distearylphosphatidylglycerol (DSPG), dioleylphosphatidylglycerol (DOPG), yolk phosphatidylglycerol (yolk) PG), natural or synthetic phosphatidylglycerols (PG), such as hydrogenated egg yolk phosphatidylglycerol, dilauroylphosphatidylinositol (DLPI), dimyristylphosphatidylinositol (DMPI), dipalmitoylphosphonate Atidyl inositol (DPPI), distearoyl phosphatidyluinositol (DSPI), dioleoyl phosphatidyluinositol (D0PI), soy phosphatidylinositol (soy PI), hydrogenated soy Phosphatidylinos Shown over natural or synthetic phosphatidyl Achijirui Roh sheet Torr such as Le a (PI). These can be used alone or as a mixture of two or more.o

As the neutral phospholipids in the present invention, soybean phosphatidylcholine, egg yolk phosphatidylcholine, hydrogenated soybean phosphatidylcholine, hydrogenated egg yolk phosphatidylcholine, dimyristylphosphatidylcholine (DMPC) , Dipalmitoylphosphatidylcholine (DPPC), dilauroylphosphatidylcholine (DLPC), distearoylphosphine Natural or synthetic phosphatidyl cholesterol such as atizircolin (DSPC), myristol palmitoyl phosphatidylcholine (MPPC), palmitoyl stearyl phosphatidylcholine (PSPC), dioleoyl phosphatidylcholine (D0PC), etc. Linnes (PC), soybean phosphatidylethanolamine, egg yolk phosphatidylethanolamine, hydrogenated soybean phosphatidylethanolamine, hydrogenated egg yolk phosphatidylethanolamine, dimiristoylphosphatidylethanolamine Min (DMPE), Dipalmitoylphosphatidylethanolamine (DPPE), Dilauroylphosphatidylethanolamine (DLPE), Distearoylphosphatidylethanolamine (DSPE), Myristylvalmitoylphosphamine Jiruetano Rua Mi emissions (MPPE), Nono. Natural or synthetic phosphatidylethanolamines (PE) such as Lumitoylstearoylphosphatidylethanolamine (pSPE) and dioleoylphosphatidylethanolamine (DOPE) can be exemplified. And the like can be used alone or in combination of two or more.

The liposome membrane of the present invention is formed by using the above-mentioned acidic phospholipid alone as a constituent component, or using the above-mentioned neutral phospholipid and an acidic phospholipid in combination according to a conventional method. Here, the combined ratio of acidic phospholipids is about 0.1 to It is preferably about 100 mol%, preferably about 1 to 90 mol%, and more preferably about 10 to 50 mol%.

The K ratio of the basic drug and the acidic phospholipid in the liposome dispersion liquid is about 0.5 to 100 equivalents of the basic drug and preferably about 1 to 6 equivalents of the basic drug. It is preferably about 0 equivalents, more preferably about 1.5 to 20 equivalents.

As the basic drug to be retained, conventionally known ones can be widely used as long as the drug is a basic drug.For example, use is made of a basic drug such as carteolol, propoterol, buprenorphine and the like and an acid addition salt thereof. Can be. The acid addition salt is not particularly limited and can be appropriately selected from a wide range. Representative examples thereof include, for example, inorganic acid salts such as hydrochloride, sulfate and phosphate, and organic acid salts such as oxalate, maleate and citrate.

Next, the method for producing the ribosome of the present invention will be described. In producing the ribosome of the present invention, various known methods can be used. For example, after dissolving the ribosome membrane constituents in an organic solvent such as macropore form, the solvent is distilled off under reduced pressure to form a lipid film, and an aqueous phase in which the drug is dissolved is added thereto, and the phase transition of lipids is performed. Heat to above temperature and prepare ribosome dispersion by vortex, homogenization, etc. In addition, the powdered ribosome membrane constituents are heated above the phase transition temperature. However, a ribosome dispersion can also be prepared by mixing and stirring with an aqueous drug solution. The drug aqueous solution to be added can be any one as long as the drug is dissolved, and the amount of the drug aqueous solution to be added can be arbitrarily increased or decreased.

If necessary, the particle size distribution of the liposome dispersion obtained in this manner can be controlled by the ultra-permeable membrane method, for example, using a membrane filter made of polycarbonate. . It is also possible to concentrate using a dialysis membrane.

In addition, various known substances such as a preservative, an isotonic agent, a buffer, a stabilizer, a solubilizer, and an absorption enhancer are appropriately added to the ribosome dispersion as additives necessary for the formulation design. It can be blended, and if necessary, can be diluted with a liquid or water containing these additives. Specific examples of the above additives include preservatives that are effective against fungi and bacteria such as benzalkonium chloride, benzethonium chloride, black hexidine, parabens (methylparaben, ethylparaben, etc.), thimerosal, etc. Preservatives, tonicity agents: polyhydric alcohols such as D-mannitol, D-sorbitol, D-xylitol, glycerin, glucose, monetose, sucrose, propylene glycol, and sodium chloride Electrolytes such as aluminum, and stabilizers such as tocopherol, butylhydroxyanisole, butylhydroxytoluene, and ethylenediaminetetranate. (EDTA), cysteine, etc., respectively.

The liposomal preparation of the present invention exhibits a high trap rate of the drug, which is not found in conventional liposomal preparations, and is excellent as a ribosome preparation which stably maintains its retention ability. It does not use irritating anionic surfactants such as sodium lauryl sulfate (SDS) and dicetyl phosphate, and is also excellent in that it has virtually no irritation. Things. In particular, the preparation of the present invention is highly useful as a pharmaceutical agent such as eye drops, nasal drops, injections and inhalants.

Example

Examples, control examples and test examples are listed below, but these examples do not limit the present invention.

Example 1

Hydrogenated soy phosphatidylcholine (PC purity 90% or more) 0.707 g (0.9 mimol), DMP G (PG purity

0.069 g (0.1 millimol) was weighed out and dissolved in a round-mouthed form 20 »ί in an eggplant-shaped flask. The solvent was distilled off using a rotary evaporator. The solvent was completely removed by suction with a vacuum pump. A 0.05% aqueous solution of carteolol hydrochloride was added to the resulting residue.

(0.03 mimol) was added, and the mixture was heated at 70 to sufficiently swell. Vortex while maintaining the temperature at 70 After sufficiently stirring, the mixture was cooled to room temperature to obtain a milky white ribosome dispersion.

Example 2

Weigh out 0.335 g (0.45 mmol) of hydrogenated soybean phosphatidylcholine (PC purity 90% or more) and 0.035 g (0.05 mol) of DMP G (PG purity 99% or more). After dissolving the solvent in a round-mouthed form 20 in an eggplant-shaped flask, the solvent was distilled off by a rotary evaporator, and the solvent was completely removed by suction with a vacuum pump. The resulting residue was added to a 0.05% aqueous carteolol hydrochloride solution.

(0.03 mimol) was added, and the mixture was heated at 70 to sufficiently swell. The mixture was sufficiently vortexed while maintaining the temperature at 70, and then cooled to room temperature to obtain a milky white ribosome dispersion.

Example 3

Hydrogenated soy phosphatidylcholine (PC purity 90% or more) 1.413 g (1.8 millimol), DMP G (PG purity 99% or more) 0.138 g (0.2 millimol) After weighing and dissolving in a round-mouth form 20 in an eggplant-shaped flask, the solvent was distilled off by a rotary evaporator, and the solvent was completely removed by suction with a vacuum pump. A 0.05% aqueous solution of carteolol hydrochloride 2 was added to the resulting residue. (0.03 milliliters) was added, and the mixture was heated at 70 to sufficiently swell. After sufficiently stirring with a vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky ribosome dispersion.

Example 4

Hydrogenated soy phosphatidylcholine (PC purity 90% or more) 0.746 g (0.95 mimol), DMPG (PG purity 9.9% or more) 0.035 g (0.05 (Millimol) was weighed out, dissolved in a closed mouth form 20 »in an eggplant-shaped flask, the solvent was distilled off by a rotary evaporator, and the solvent was completely removed by suction with a vacuum pump. To the resulting residue was added 0.05% aqueous carteolol hydrochloride aqueous solution 20 (0.03 mmol), and the mixture was heated to 70 to sufficiently swell. After sufficiently stirring with vortex while keeping the humidity at 70, the mixture was cooled to room temperature to obtain a milky ribosome dispersion.

Example 5

Hydrogenated soy phosphatidylcholine (PC purity 90% or more) 0.50 g (0.7 millimol), DMP G (PG purity

0.29 g <0.3 millimol) was weighed and dissolved in a round-mouthed form 20 J in an eggplant-shaped flask, and then the solvent was distilled off using a rotary evaporator. More vacuum The solvent was completely removed by suction with a pump. A 0.05% aqueous solution of carteolol hydrochloride 20% was added to the resulting residue.

(0.03 mimol) was added, and the mixture was heated to 70 to sufficiently swell. After the mixture was sufficiently stirred by vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white liposome dispersion.

Example 6

A mixture of hydrogenated soybean phosphatidylcholine (PC purity of 90% or more) and 10 mol% of DMP G (PG purity of 99% or more) was added to 0.776 g (1.0 millimol) of lipid powder. Then, a 20% (0.03 mimol) of a 0.05% protic terol aqueous solution of hanic acid was added, and the mixture was heated at 70 to sufficiently swell. After sufficiently stirring with a vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white ribosome dispersion. Example 7

In advance, 0.776 g (1.0 millimol) of lipid powder obtained by adding 10 mol% of DMP G (PG purity of 99% or more) to hydrogenated soybean phosphatidylcholine (PC purity of 90% or more) was added. A 0.05% aqueous solution of buprenorphine hydrochloride (20 ar, 0.02 mimol) was added, and the mixture was heated at 70 to sufficiently swell. After sufficiently stirring with vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white ribosome dispersion. Example 8

To 0.5 g of lipid powder obtained by adding 30 mol% of DPPG (PG purity of 9.9% or more) to DPPC (PC purity of 9.9% or more) in advance, add 0.2% 1% buprenorphine hydrochloride aqueous solution. 9.5 (0.04 mmol) was added, and the mixture was heated at で 0 to sufficiently swell. After sufficiently stirring with vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white 0.2% buprenorphine hydrochloride isotonic liposome dispersion.

Example 9

0.777 g (0.99 mimol) of hydrogenated soybean phosphatidylcholine and 0.07 g (0.01 mimol) of DPPG are weighed out, and they are closed in an eggplant-shaped flask. After dissolving in form 20, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.05% aqueous solution of propoterol hydrochloride containing 2.5% glycerin, and 203 ° (0.003 mmol) was added. Swelled well. Temperature

The mixture was stirred at 70 ° C. and cooled to room temperature to obtain a milky white liposome dispersion.

Example 10

Weigh 0.77 g (0.9 mimol) of hydrogenated soybean phosphatidylcholine and 0.07 g (0.1 mimol) of DPPG. The solution was taken out and dissolved in a round-mouthed form 20 in an eggplant-shaped flask, and then the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.05% aqueous solution of procaterol hydrochloride 2 One (0.03 mimol) containing 2.5% glycerin, and the mixture was heated to 70 to sufficiently swell. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 1 1

Take 0.393 g (0.5 mmol) of hydrogenated soybean phosphatidylcholine and 0.352 g (0.5 mmol) of DPPG and dissolve them in 20% of the form in a round-mouthed flask in an eggplant-shaped flask. After squeezing, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.2% 25% aqueous solution of propower terol hydrochloride containing 2.3% glycerin, and the mixture was heated to 70 to sufficiently moisten the solution. . While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 1 2

Take 0.079 g (0.1 mimol) of hydrogenated soybean phosphatidylcholine and 0.633 g (0.9 mimol) of DPPG, and dissolve them in 20 ml of black-mouthed form in an eggplant-shaped flask. After squeezing, the solvent was distilled off under reduced pressure to completely remove the solvent. Profit To the residue thus obtained was added 0.45% aqueous solution of propoterol hydrochloride 2 (0.27 mmol) containing 2.2% glycerin, and the mixture was heated at 70 to sufficiently swell. 70 to the temperature. While stirring at C, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 13

0.73 g (1 mmol) of DPPG is weighed, and 0.5% aqueous solution of procapteroyl hydrochloride containing 2.2% glycerin is added, and 20 π & (3 mmol) is added. The mixture was warmed to swell sufficiently. And cooled to room temperature and stirred while the temperature was kept at 7 0 ^e C, to obtain a milky white ribosome dispersion.

Example 14

DMP C 0.672 g (0.999 mimol) and

Weigh out 0.007 g (0.01 mimol) of DPPG, dissolve it in 20 mL of porcelain form in a NAS flask, and then distill off the solvent under reduced pressure to completely remove the solvent. did. To the obtained residue was added a 0.05% aqueous solution of procaterol hydrochloride containing 2.5% glycerin (0.03 milliliters), and the mixture was heated to 70 and swelled sufficiently. I was sorry. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion. After weighing DMP C O. 61 1 g (0.9 millimol) and DPPG 0.07 g (0.1 millimol), dissolve them in a round-mouthed form 20 »in an eggplant-shaped flask. The solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.05% aqueous solution of propoterol hydrochloride containing 2.3% glycerin (20% (0.03 mimol)), and the mixture was heated at 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 16

Take 0.34 g (0.5 mimol) of DMP C and 0.35 g (0.5 mimol) of DPPG, dissolve them in 20 mL of form in a round-bottomed flask, and remove the solvent. The solvent was removed under reduced pressure to completely remove the solvent. To the obtained residue was added 20 ^ (0.15 mimol) of a 0.25% aqueous solution of propowerol hydrochloride containing 2.2% glycerin, and the mixture was heated at 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 17

Take out 0.68 g (0.1 mmol) of DMP C and 0.633 g (0.9 mmol) of DPPG, dissolve them in black-mouthed form 20 in an eggplant-shaped flask, and remove the solvent. Was distilled off under reduced pressure to completely remove the solvent. 2.5% in the residue obtained A 0.25% (0.27 millimol) aqueous solution of propoterol hydrochloride containing 0.45% glycerin was added, and the mixture was heated to 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 18

0.668 g (0.85 mimol) of hydrogenated soybean phosphatidylcholine and 0.105 g (0.149 mimol) of DPPG were weighed and placed in an eggplant-shaped flask. After dissolving in form 20, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.5% aqueous solution of propoterol hydrochloride containing 2.5% glycerin, 20 * (0.298 millimol), and the mixture was heated to 70 to sufficiently swell. Was. Temperature

The mixture was stirred and cooled to room temperature while keeping at 70 to obtain a milky white liposome dispersion.

Example 19

0.55 g (0.7 mimol) of hydrogenated soybean phosphatidylcholine and 0.29 g (0.3 mimol) of DPPG were weighed and placed in a round bottom form 20 in an eggplant-shaped flask. After dissolution, the solvent was distilled off under reduced pressure to completely remove the solvent. A 0.5% aqueous solution of procaterol hydrochloride containing 2.3% glycerin was added to the obtained residue, and 20% (0.298 millimol) of the solution was added, and the mixture was heated to 70 to sufficiently swell. . Keep the temperature at 70 The suspension was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 20

Take 0.317 g (0.4 mmol) of hydrogenated soybean phosphatidylcholine and 0.419 g (0.6 mmol) of DPPG and dissolve them in a closed mouth form 20 in an eggplant-shaped flask. After squeezing, the solvent was distilled off under reduced pressure to completely remove the solvent. A 0.5% aqueous solution of procaterol hydrochloride containing 2.2% glycerin in 20% (0.298 millimol) was added to the obtained residue, and the mixture was heated to 70 to sufficiently swell. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 2 1

0,084 g (0.11 mimol) of hydrogenated soybean phosphatidylcholine and 0.628 g (0.89 mimol) of DPPG were weighed out and dissolved in 20 ml of form-cloth in an eggplant-shaped flask. Thereafter, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.3% aqueous solution of propowererol hydrochloride 2 (0.18 mmol) containing 2.3% glycerin, and the mixture was heated at 70 to sufficiently swell. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a white ribosome dispersion. Example 22

Weigh out 0.645 g (0.82 mimol) of hydrogenated soybean phosphatidylcholine and 0.126 g (0.18 mimol) of DPPG and dissolve in black-mouthed form 20 in an eggplant-shaped flask. After that, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.03% aqueous solution of propotterol hydrochloride containing 2.2% glycerin (0.018 millimoles), and the mixture was heated to 70 to swell sufficiently. Was. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white liposome dispersion.

Example 23

Weigh out 0.368 g (0.5 mmol) of DPPC and 0.352 g (0.5 mmol) of DPPG, dissolve in Cloth Form 2 in an eggplant-shaped flask, and depressurize the solvent. The solvent was distilled off to completely remove the solvent. To the obtained residue was added a 0.05% aqueous solution of propoterol hydrochloride 20 (0.03 mmol) containing 2.5% glycerin, and the mixture was heated to 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 24

0.396 g (0.5 mimol) of DSPC and 0.380 g (0.5 mimol) of DSPG were weighed, and the eggplant type flask was weighed. After dissolving 20 ml of the form in the flask, the solvent was distilled off under reduced pressure to completely remove the solvent. To the resulting residue was added a 0.05% aqueous solution of propoterol hydrochloride (0.03 mimol) containing 2.5% glycerin, and the mixture was heated to 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 25

0.393 g (0.5 mimol) of hydrogenated soybean phosphatidylcholine and hydrogenated soybean phosphatidylinositol

0.41 g (0.5 millimol) was weighed out, dissolved in black-mouthed form 20 ^ in an eggplant-shaped flask, and then the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added a 0.05% aqueous solution of propowererol hydrochloride 20 (0,03 mmol) containing 2.5% glycerin, and the mixture was heated at 70 to swell to a sufficient extent. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 26

0.336 g (0.5 mmol) of DPPE and 0.352 g (0.5 mmol) of DPPG were weighed out, dissolved in Cloth Form 2 in an eggplant-shaped flask, and the solvent was removed. Was distilled off under reduced pressure to completely remove the solvent. The resulting supernatant contains 2.5% glycerin 0.05% proproterol hydrochloride aqueous solution 20 (0.03 mimol) was added, and the mixture was heated to 70 ° C to swell sufficiently. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Example 2 7

0.368 g (0.5 mmol) of DPPC and 0.352 g (0.5 mmol) of DPPG were weighed, and dissolved in black-mouthed form 20 in an eggplant-shaped flask. Thereafter, the solvent was distilled off under reduced pressure to completely remove the solvent. To the obtained residue was added 20% (0.03 mimol) of an aqueous solution of 2.5% glycerin containing 0.05% glycerol, and the mixture was heated to 70% to fill. Swelled. While maintaining the temperature at 70, the mixture was stirred and cooled to room temperature to obtain a milky white ribosome dispersion.

Control Example 1

0.785 g (1 mmol) of hydrogenated soybean phosphatidylcholine (PC purity: 90% or more) was weighed out, dissolved in a round-mouthed form 20 * in an eggplant-shaped flask, and then rotary evaporated. The solvent was distilled off using a porator, and the solvent was completely removed by suction using a vacuum pump. To the obtained residue, 0.05% aqueous carteolol hydrochloride aqueous solution 20 was added, and the mixture was heated at 70 to sufficiently swell. After sufficiently stirring with vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white liposome dispersion. Control 2

Powder of hydrogenated soy phosphatidylcholine (PC purity 90% or more)

0.785 g (1 mimol) was weighed, and a 0.05% aqueous solution of procaterol hydrochloride 2 O J ^ was added, and the mixture was heated to 70 to sufficiently swell. The mixture was sufficiently stirred with a vortex while maintaining the temperature at 70, and then cooled to room temperature to obtain a milky white liposome dispersion.

Control 3

0.785 g (1 mmol) of powdered hydrogenated soybean phosphatidylcholine (PC purity of 90% or more) was weighed, and 0.05% buprenorphine hydrochloride aqueous solution 20 »was added. Heated to 0 and swelled sufficiently. After sufficiently stirring with vortex while maintaining the temperature at 70, the mixture was cooled to room temperature to obtain a milky white liposome dispersion.

Control Example 4

A 0.2% buprenorphine hydrochloride isotonic aqueous solution was prepared. Control 5

Hydrogenated soy phosphatidylcholine (PC purity 90% or more) 0.707 g (0.9 millimol), DPPA (PA purity)

0.90 g (0.1 mimol) was taken out and dissolved in a round-mouthed form 203 ^ in an eggplant-shaped flask. The solvent was distilled off using a rotary evaporator, and the solvent was completely removed by suction using a vacuum pump. To the obtained first residue was added 0.05% (0.03 mimol) of a 0.05% aqueous carteolol hydrochloride solution, and the mixture was added to the residue. C was heated and swelled sufficiently. After sufficiently stirring with a vortex while keeping the temperature at 70, the mixture was cooled to room temperature to obtain a milky white liposome dispersion.

Control 6

DMP C (PC purity 98% or more) 0.62 g (0.9 millimol amount), DMP S (PS purity 9.9% or more)

0.069 g (0.1 millimol) was taken out, dissolved in a black-mouthed form 20 »^ in an eggplant-shaped flask, and then the solvent was distilled off by a rotary evaporator. More suction was applied to completely remove the solvent. A 0.05% aqueous solution of carteolol hydrochloride 20 β was added to the obtained paste-like residue.

(0.03 mimol) was added, and the mixture was heated to 70 to sufficiently swell. The mixture was sufficiently vortexed while maintaining the temperature at 70, and then cooled to room temperature to obtain a milky white liposome dispersion.

Test example 1

For the ribosome dispersions obtained in Examples 1, 2, 3, 4, and 5 and Control Examples 1, 5, and 6, Carteolol hydrochloride was used. Was measured. The results are shown in Table 1.

Table 1

From Test Example 1, the ribosome preparation of the present invention shows that the amount of the ribosome membrane component in the ribosome dispersion, the types of neutral phospholipids and acidic phospholipids in the ribosome membrane component, and the concentration of the acidic phospholipid are different. It can be seen that even when changed, it shows good drug retention ability. On the other hand, the preparations of Control Examples 5 and 6 using phosphatidylserine and phosphatidic acid, respectively, still have a lower trapping rate of carteolol hydrochloride than the drug using the acidic phospholipid of the present invention. It turns out to be insufficient.

Test example 2

The liposome dispersions obtained in Example 6 and Control Example 2 were Then, the trapping rate of proforcerol hydrochloride was measured. Table 2 shows the results.

Table 2

Test Example 2 shows that the liposome preparation of the present invention prepared by various methods exhibits excellent drug-retaining ability even when the drug is proforceterol hydrochloride.

Test example 3

With respect to the liposome dispersions obtained in Example 7 and Control 3, the trapping rate of buprenorphine hydrochloride was measured. The results are shown in Table 3.

Table 3

Test Example 3 shows that the liposome preparation of the present invention prepared by various methods shows good drug retention ability even when the drug is buprenorphine hydrochloride. Test example 4

With respect to the liposome dispersions obtained in Example 1 and Control Example 1, the trapping rate of the carteohydrochloride port immediately after preparation and after 17 days of storage at room temperature was measured and compared. Table 4 shows the results.

Table 4

Test Example 4 shows that the ribosome preparation of the present invention does not show a decrease in drug retention ability over time and is stable over time.o

Test example 5

The liposome dispersions obtained in Example 1 and Control Example 1 were subjected to steam sterilization under pressure (121, 40 minutes), and the trapping rate of luteolol was measured. Table 5 shows the results. Five

Test Example 5 shows that the ribosome preparation of the present invention does not show a decrease in drug retention ability due to pressurized steam sterilization (121, 40 minutes) and is thermodynamically stable.

As is clear from these results, the liposomal preparation containing the acidic phospholipid of the present invention shows a remarkably high trapping rate for basic drugs as compared with the conventional liposome preparations, It is an excellent ribosome preparation that maintains its retention ability over time and thermodynamically.

Test example 6

With respect to the ribosome dispersions obtained in Examples 1 and 8 and Control Example 1, each of them was diluted 10-fold with the use of PBS isotonic buffer at 37 and calf serum, respectively. Made. Each of these solutions was incubated at 37, and the rate of trapping of carteolol hydrochloride from the ribosome over time was measured. The results are shown in Table 6 (trapping rate of carteolol hydrochloride in PBS isotonic buffer) and Table 7 (calf blood Table 6 shows the percentage of carteolol hydrochloride trapped in Qing

From Test Example 6, the ribosome preparation of the present invention The morphology was maintained for a long time even in PBS isotonic buffer and calf serum, indicating that the drug retention ability did not decrease over time and was stable.

Test example 7

200 of each of the liposome dispersion obtained in Example 8 and the aqueous solution of Control 4 were applied to the thigh of a rat (SD strain, male, weight of about 200 g) in each group of 3 animals per group. injected in muscles, the plasma hydrochloride Bupureno Ruch fin density of each rat (n _S / M £) was measured over time. The results obtained are shown in Table 8 as the average value of three rats in each group.

Table 8

Example 8 Control 4

20 minutes later 26. 0 287.2

40 minutes later 3 1.5 1 7 1.8

1 hour later 42. 7 1 25.5

2 hours later 3 1.2 27.1

4 hours later 38, 0 31.5

6 hours later 29.3 18.8

8 hours later 19.8 15.3

After 10 hours 1 7.5 14.5

24 hours later 1.7.4.5.7

48 hours later 1. 47, 3 As is clear from the results of the above test examples, all of the liposomal preparations of the present invention show a remarkably high trapping rate for basic drugs as well as the drug retention compared to conventional ribosome preparations. Ability can be stably maintained over a long period of time, and the retention ability is thermodynamically stable. In addition, the liposomal preparation of the present invention does not rapidly lose the above-mentioned drug-retaining ability even in a living body, and is therefore effective as a means for an aqueous sustained-release preparation.

Claims

0 Q

O 91/14423 "PCT / JP91 / 00373

The scope of the claims

① Use acidic phospholipids as membrane components or neutral phospholipids

A liposome preparation comprising a liposome containing a basic drug in a liposome, and a membrane component consisting of liposome and acidic phospholipid.

② 0.5 to 100 equivalents of acidic phospholipids for basic drugs

The liposome preparation according to claim 1 used in an amount,

(3) The use of neutral phospholipids and acidic phospholipids as membrane components

3. The liposome preparation according to item 2 of the claim.

The liposomal preparation according to claim 2, wherein the acidic phospholipid is a natural or synthetic phosphatidylglycerol.

が Neutral phospholipids are natural or synthetic phosphatidylcholines

4. The liposome preparation according to claim 3, which is:

リ The liposome preparation according to claim 5, wherein the acidic phospholipid is a natural or synthetic phosphatidyl glycerol, ⑦ the basic drug is carteolol, proproterol or

Claims 2 to 4 selected from Brenorphin

7. The liposome preparation according to any one of the items up to 6.

項 Claim 2 wherein the basic drug is propoterol

7. The liposome preparation according to any one of to 6.