METHOD AND APPARATUS FOR PREPARING NOVEL LIPOSOME

• US 20130069261A1
• Filed: 02/24/2011
• Published: 03/21/2013
• Est. Priority Date: 02/24/2010
• Status: Active Grant

• Current Assignee: BCWorld Pharm Co., Ltd.
• Original Assignee: Sung Joo Hwang
• Inventors: Hwang, Sung Joo, Park, Hee Jun, Cho, Wonkyung, Cha, Kwang-Ho, Park, Junsung, Park, Chanhyuk, Gu, Donggeon
• Granted Patent: US 9,579,261 B2
• Time in Patent Office: Days
• US Class Current: 264/4.1
• CPC Class Codes: A61J 3/07 into the form of capsules or similar small containers for oral use
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: A61K 9/19 lyophilised , i.e. freeze-dried, solutions or dispersions lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: B01D 11/0411 the supercritical fluid acting as solvent for the solvent and as anti-solvent for the solute, e.g. formation of particles from solutions

Abstract

Disclosed is a method for preparing a liposome formulation. In the disclosed method, a lipid fraction is dissolved in an organic solvent. The solution including a bioactive component and the lipid fraction, together with a carrier, is put in a reaction vessel, and a supercritical fluid is introduced thereto, so as to prepare particles coated with the bioactive component-lipid. The supercritical fluid is discharged by compression to obtain proliposome particles, and then the proliposome particles are hydrated by an aqueous solution including water so as to form a liposome solution. Preferably, the formulation may include one or more bioactive components. As required, the liposome formulation may be further processed by methods such as particle size reduction, removal of organic solvent, and freeze-drying. The preparation method can be easily carried out at a laboratory scale. Furthermore, the same method can be employed in liposome formulation preparation in mass production, or at a commercial scale.

Claims

1. A method for preparing liposome formulation, the method comprising the steps of:

(a) dissolving a lipid fraction comprising at least one kind selected from phospholipid and sterol, and a bioactive substance in an organic solvent;

(b) putting an organic solvent solution of the step (a) together with a saccharide carrier into a reaction vessel, and introducing a supercritical fluid thereto so as to prepare proliposome particles coated with a lipid-bioactive substance mixture;

(c) discharging the supercritical fluid in step (b) by compression so as to obtain the proliposome particles; and

(d) hydrating the proliposome particles in step (c) by an aqueous solution comprising water so as to form a liposome solution.

2. A method as claimed in claim 1, further comprising the step of reducing a liposome particle size by passing the liposome solution in step (d) through a microfluidizer.

3. A method as claimed in claim 1, wherein the phospholipid is at least one kind selected from the group including phosphatidyl glycerols, phosphatidyl cholines, phosphatidyl ethanolamines, phosphatidylserines, phosphatidylinositols, yolk lecithin, soybean lecithin, N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methylsulfate (DOTAP), N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), dioctadecyldimethylammonium bromide (DODAB), 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate (DOSPA), dioleoyl phosphatidylethanolamine (DOPE), 1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide (DMRIE), didodecyldimethylammonium bromide (DDAB), 3β

-[N—

(N′

,N′

-dimethylaminoethane)-carbamoyl]cholesterol (DC-Chol), dioctadecylamidoglycylspermine (DOGS), N,N-[bis(2-hydroxyethyl)]-N-methyl-N-[2,3-di(tetradecanoyloxy)propyl]ammonium iodide, [N,N,N′

,N′

-tetramethyl-N,N′

-bis(2-hydroxyethyl)-2,3-di(oleoyloxy)-1,4-butanediammonium iodide], diethylaminoethyl cellulose (DEAE-C), N,N,N,N-tetramethyl-N,N,N,N-tetrapalmitylspermine, dioleoyl phosphatidylethanolamine, N-t-butyl-N′

-tetradecyl-3-tetradecylaminopropionamidine, and diethylaminoethyl dextran (DEAE-D).

4. A method as claimed in claim 1, wherein the sterol is at least one kind selected from the group including cholesterol, cholesterol hexasuccinate, ergosterol, and lanosterol.

5. A method as claimed in claim 1, wherein the organic solvent is at least one kind selected from the group including dimethylacetamide, dimethylformamide, dimethylsulfoxide, chloroform, methanol, ethanol, and ether.

6. A method as claimed in claim 1, wherein the bioactive substance is at least one kind selected from the group including autonomic nerve modulators, corticosteroids, diuretics, analgesics, sex hormones, anesthetics, anthelminitics, antihistamines, antiprotozoal agents, antianemics, cardiovascular disease agents, antianxiety agents, antiasthmatics, anticonvulsants, antidepressants, antidiabetics, antidiuretics, antidotes, antiepileptics, antifungal agents, antihyperlipidemic agents, antihyperlipoproteinemics, antihypertensives, antihypotensives, antibiotics, analgesics, antimigraines, antimycotics, antinauseants, anticancer agents, antiparkinsonians, antipsoriatics, neuropsychiatric agents, antiplatelet agents, antitussive expectorant agents, anti-ulceratives, bronchodilators, cardiotonics, diuretics, emetics, anti-ulceratives, hormones, antihyperlipidemic agents, immunomodulators, muscle relaxants, neuroleptics, vasodilators, antivirals, insecticides, protein drugs, gene drugs and antibodies.

7. A method as claimed in claim 6, wherein the bioactive substance is amphotericin B.

8. A method as claimed in claim 1, wherein the saccharide carrier is at least one kind selected from the group including lactose, sucrose, maltose, trehalose, dextrose, sorbitol, mannitol, and xylitol.

9. A method as claimed in claim 1, wherein the supercritical fluid is selected from the group including supercritical carbon dioxide, supercritical nitrogen monoxide, supercritical acetylene, supercritical trifluoromethane, supercritical propane, supercritical ethylene, supercritical chlorofluocarbon and supercritical xenon.

10. A method as claimed in claim 1, further comprising the step of sterile-filtrating the prepared liposome.

11. A method as claimed in claim 1, further comprising the step of freeze-drying the prepared liposome.

12. An apparatus for performing the method as claimed in claim 1, the apparatus comprising:

a reaction vessel in which the step of preparing proliposome particles and the step of forming a liposome solution are carried out;

a supercritical fluid storage vessel for receiving a supercritical fluid;

a supercritical fluid supplying tube for transporting the supercritical fluid to the reaction vessel from the supercritical fluid storage vessel, which is connected from the supercritical fluid storage vessel to the reaction vessel;

a supercritical fluid supply amount adjusting pump for controlling an amount of the transported supercritical fluid, which exists in the supercritical fluid supplying tube;

a reactive material introducing part for providing a reactive material to the reaction vessel, which is connected to inside of the reaction vessel;

a pressure meter for measuring a pressure in the reaction vessel, which is connected to the inside of the reaction vessel;

a supercritical fluid discharging tube for discharging the supercritical fluid, which is connected to the reaction vessel;

a pressure-reducing valve for discharging the supercritical fluid, which exists in the supercritical fluid discharging tube; and

a back pressure valve for discharging the supercritical fluid while maintaining a predetermined pressure or more, which exists in the supercritical fluid discharging tube.

13. An apparatus as claimed in claim 12, wherein an agitating device exists within the reaction vessel, and the reaction vessel comprises an agitator in itself or at outside thereof.