US4619794A
Expired / Lapsed
US
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Spontaneous preparation of small unilamellar liposomes
Assignee: Novartis Corp
•
Filing: 1985-01-29
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Granted: 1986-10-28
•
Expires: 2005-01-29
Abstract
Abstract The present invention relates to a novel advantageous process for the preparation of unilamellar liposomes in aqueous phase by converting a suitable lipid component, e.g. phosphatidic acid, into the ionic form by subjecting the lipid dispersion to a change in pH value and subsequently neutralizing it. Formation of the unilamellar liposomes is spontaneous, i.e. it takes place without additional external supply of energy. The liposomes obtainable by the process of this invention can be used therapeutically as carriers for drugs of the most widely different kind.
Inventors
- 👤 Helmut Hauser
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 02:00
CPC Classifications
A; A61; A61K; A61K9/00; A61K9/10; A61K9/127; A61K9/1271; A61K9/1272; A61K9/1277; Y; Y10; Y10S; Y10S436/00; Y10S436/829; Y10T; Y10T428/00; Y10T428/29; Y10T428/2982; Y10T428/2984
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
797 words
A process for the spontaneous preparation of small unilamellar liposomes comprising, without additional energy input, dispersing (a) a homogeneous layer of a phospholipid of the formula ##STR2## wherein m is zero or one, one of R 1 and R 2 is hydrogen, hydroxy or C 1 -C 4 -alkyl and the other is alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl, and R 4 is hydrogen, C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3- amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-tri-methylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms, or, if R 1 and R 2 are hydrogen or hydroxy and R 3 is hydrogen, is a steroid radical, and an additional phospholipid of the formula A, wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms; or of a phospholipid of the formula A, wherein m is zero or one, each of R 1 and R 2 independently of the other is alkyl, alkenyl, alkoxy, or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, and R 3 and R 4 are hydrogen and optionally an additional phospholipid of the formula A wherein m is zero or one, each of R 1 and R 2 independently of the other is alkyl, alkenyl, alkoxy, or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, and R 3 is hydrogen or C 1 -C 4 -alkyl, and R.sub. 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms in an aqueous phase which is raised to a pH-value at least 11 or (b) a homogeneous layer of a phospholipid of the formula A wherein m is 0 or 1, one of R 1 and R 2 is hydrogen, hydroxy or C 1 -C 4 -alkyl, and the other is alkyl, alkenyl, alkoxy or alkenyloxy, each of 10 to 20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, R 3 is hydrogen, and R 4 is lower alkyl which is substituted by an ammonio group and an additional phospholipid of the formula A wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2- carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylene- dioxypropyl, 2,3-(2,2-propylene) dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms; or of a phospholipid of the formula A, wherein m is 0 or 1, each of R 1 and R 2 independently of the other is alkyl, alkenyl or alkenyloxy, each of 10 to 20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, R 3 is hydrogen, and R 4 is lower alkyl which is substituted by an ammonio-lower alkylammonio group and an additional phospholipid of the formula A wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenoxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyhl, or a carbohydrate radical of 5 to 12 carbon atoms, in an aqueous phase which is lowered to a pH-value of about or less than about 1 and the aqueous phase is neutralized with a physiologically acceptable acid or base and the unilamellar liposomes obtained are separated from multilamellar liposomes.
Claim 2
Dependent Claim
797 words
A process for the spontaneous preparation of small unilamellar liposomes comprising, without additional energy input, dispersing (a) a homogeneous layer of a phospholipid of the formula ##STR2## wherein m is zero or one, one of R 1 and R 2 is hydrogen, hydroxy or C 1 -C 4 -alkyl and the other is alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl, and R 4 is hydrogen, C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3- amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-tri-methylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms, or, if R 1 and R 2 are hydrogen or hydroxy and R 3 is hydrogen, is a steroid radical, and an additional phospholipid of the formula A, wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms; or of a phospholipid of the formula A, wherein m is zero or one, each of R 1 and R 2 independently of the other is alkyl, alkenyl, alkoxy, or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, and R 3 and R 4 are hydrogen and optionally an additional phospholipid of the formula A wherein m is zero or one, each of R 1 and R 2 independently of the other is alkyl, alkenyl, alkoxy, or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, and R 3 is hydrogen or C 1 -C 4 -alkyl, and R.sub. 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms in an aqueous phase which is raised to a pH-value at least 11 or (b) a homogeneous layer of a phospholipid of the formula A wherein m is 0 or 1, one of R 1 and R 2 is hydrogen, hydroxy or C 1 -C 4 -alkyl, and the other is alkyl, alkenyl, alkoxy or alkenyloxy, each of 10 to 20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, R 3 is hydrogen, and R 4 is lower alkyl which is substituted by an ammonio group and an additional phospholipid of the formula A wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenyloxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2- carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylene- dioxypropyl, 2,3-(2,2-propylene) dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms; or of a phospholipid of the formula A, wherein m is 0 or 1, each of R 1 and R 2 independently of the other is alkyl, alkenyl or alkenyloxy, each of 10 to 20 carbon atoms, or is acyloxy of 10 to 50 carbon atoms, R 3 is hydrogen, and R 4 is lower alkyl which is substituted by an ammonio-lower alkylammonio group and an additional phospholipid of the formula A wherein each of R 1 and R 2 independently of the other are alkyl, alkenyl, alkoxy or alkenoxy, each of 10-20 carbon atoms, or is acyloxy of 10-50 carbon atoms, R 3 is hydrogen or C 1 -C 4 -alkyl and R 4 is C 1 -C 7 -alkyl, carboxy-lower alkyl, 2-aminoethyl, 3-amino-n-propyl, 2-methylaminoethyl, 3-methylamino-n-propyl, 2-dimethylaminoethyl, 3-dimethylamino-n-propyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-trimethylammonioethyl, 2-triethylammonioethyl, 3-trimethylammonio-n-propyl, 3-triethylammonio-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-methoxyethyl, 3-methoxy-n-propyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloro- or 2-bromoethyl, 2- or 3-chloro-n-propyl or 2- or 3-bromo-n-propyhl, or a carbohydrate radical of 5 to 12 carbon atoms, in an aqueous phase which is lowered to a pH-value of about or less than about 1 and the aqueous phase is neutralized with a physiologically acceptable acid or base and the unilamellar liposomes obtained are separated from multilamellar liposomes.
Claim 3
Dependent Claim
21 words
A process according to claim 1, wherein the aqueous dispersion contains egg phosphatidic acid, or egg phosphatidic acid and egg lecithin.
Claim 4
Dependent Claim
20 words
A process according to claim 1, wherein the aqueous dispersion contains egg phosphatidic acid, egg lecithin or beef brain phosphatidylserine.
Claim 5
Dependent Claim
12 words
A process according to claim 1, wherein the aqueous dispersion contains asolectin.
Claim 6
Dependent Claim
18 words
A process according to claim 1, wherein the aqueous dispersion contains egg phosphatidic acid, egg lecithin and cholesterol.
Claim 7
Dependent Claim
15 words
A process according to claim 1, wherein the aqueous dispersion contains lysolecithin and egg lecithin.
Claim 8
Dependent Claim
16 words
A process according to claim 1, wherein the aqueous dispersion contains natural lysophosphatidylserine and egg lecithin.
Claim 9
Dependent Claim
30 words
A process according to claim 1, wherein a homogeneous layer of the lipids as specified under process (a) is dispersed and the pH value is subsequently raised to about 12.
Claim 10
Dependent Claim
27 words
A process according to claim 8, wherein the pH value is raised by the addition of dilute aqueous sodium hydroxide solution or dilute aqueous potassium hydroxide solution.
Claim 11
Dependent Claim
31 words
A process according to claim 8, wherein a homogeneous layer of the lipids as specified under process (a) is dispersed in an aqueous phase having a pH value higher than 7.
Claim 12
Dependent Claim
29 words
A process according to claim 1, wherein a homogeneous layer of the lipids as specified under process (a) is dispersed in dilute aqueous sodium hydroxide or potassium hydroxide solution.
Claim 13
Dependent Claim
30 words
A process according to claim 1, wherein a homogeneous layer of the lipids as specified under process (b) is dispersed in an aqueous phase having a pH of about 1.
Claim 14
Dependent Claim
32 words
A process according to claim 1, wherein the aqueous phase is subsequently neutralised by the addition of a physiologically acceptable acid, base or buffer solution having a pH of 7 to 8.
Claim 15
Dependent Claim
30 words
A process according to claim 13, wherein the aqueous phase is neutralised by the addition of a physiologically acceptable acid or buffer solution having a pH of 7 to 8.
Claim 16
Dependent Claim
15 words
A process according to claim 14, wherein the aqueous phase is neutralised with hydrochloric acid.
Claim 17
Dependent Claim
99 words
A process according to claim 1, characterized in that a lipid of the formula A, wherein m is 1, R 1 is alkyl, R 2 is hydrogen or hydroxy, R 3 is hydrogen or lower alkyl, and R 4 is hydrogen, C 1 -C 7 -alkyl, a carbohydrate radical of 5 to 12 carbon atoms, or a steroid radical, and an additional lipid of the formula A, wherein R 1 and R 2 are acyloxy, R 3 is hydrogen, and R 4 is 2-trimethylammonioethyl, 2-aminoethyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms are dispersed.
Claim 18
Dependent Claim
109 words
A process according to claim 1, characterized in that a lipid of formula A, wherein m is 1, R 1 is n-dodecyl (lauryl), n-tetradecyl (myristyl), n-hexadecyl (cetyl), n-octadecyl (stearyl), n-dodecyloxy (lauryloxy), n-tetradecyloxy (myristyloxy), n-hexadecyloxy (cetyloxy), n-octadecyloxy (stearyloxy), lauroyloxy, myristoyloxy, 2-hydroxyethyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, or palmitoyloxy or stearoyloxy, R 2 is hydrogen or hydroxy, R 3 is hydrogen or methyl, and R 4 is hydrogen, methyl, ethyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2,3-ethylenedioxypropyl, 2,3-(2,2-propylene)dioxypropyl, 2-chloroethyl, 2-bromoethyl, or the inositol radical and an additonal lipid of formula A wherein R 1 and R 2 are lauroyloxy, myristoyloxy, palmitoyloxy, or stearoyloxy, R 3 is hydrogen and R 4 is trimethylammonioethyl or 2-aminoethyl, are dispersed.
Claim 19
Dependent Claim
51 words
A process according to claim 1, characterized in that natural lysophosphatidic acid, a synthetic lysophosphatidic acid, beef brain lysophosphatidylserine, synthetic lysophosphatidylserine, lysophosphatidylglycerol, lysophosphatidylinositol, and additionally natural lecithin, synthetic lecithin containing identical acyloxy groups, synthetic lecithin containing different acyloxy groups, or natural cephalin or synthetic cephalin containing different acyloxy groups are dispersed.
Claim 20
Dependent Claim
29 words
A process according to claim 1, characterized in that egg lysophosphatidic acid, 1-lauroyllysophosphatidic acid, 1-myristoyllysophosphatidic acid, 1-palmitoyllysophosphatidic acid, 1-myristoyllysophosphatidylserine, 1-palmitoyllysophosphatidylserine, and additionally egg lecithin 1-palmitoyl-2-oleoyllecithin, or 1-palmitoyl-2-oleoylcephalin are dispersed.
Claim 21
Dependent Claim
72 words
A process according to claim 1, characterized in that a lipid of the formula A, wherein R 1 and R 2 are acyloxy and R 3 and R 4 are hydrogen, and optionally an additional lipid of the formula A, wherein R 1 and R 2 are acyloxy, R 3 is hydrogen, and R 4 is 2-trimethylammonioethyl, 2-aminoethyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl, or a carbohydrate radical of 5 to 12 carbon atoms, are dispersed.
Claim 22
Dependent Claim
72 words
A process according to claim 1, characterized in that a lipid of formula A, wherein R 1 and R 2 are lauroyloxy, myristoyloxy, palmitoyloxy, or stearoyloxy, R 3 and R 4 are hydrogen and optionally an additional lipid of the formula A, wherein R 1 and R 2 are lauroyloxy, myristoyloxy, palmitoyloxy, or stearoyloxy, R 3 is hydrogen and R 4 is 2-trimethylammonioethyl, 2-aminoethyl, 2-amino-2-carboxyethyl, 3-amino-3-carboxy-n-propyl or the inositol radical, are dispersed.
Claim 23
Dependent Claim
47 words
A process according to claim 1, characterized in that natural phosphatidic acid or synthetic phosphatidic acid, and optionally natural lecithin, synthetic lecithin containing identical acyloxy groups, synthetic lecithin containing different acyloxy groups, natural cephalin, synthetic cephalin containing different acyloxy groups, natural phosphatidylserine, or synthetic phosphatidylserine are dispersed.
Claim 24
Dependent Claim
33 words
A process according to claim 1, characterized in that egg phosphatidic acid, dilauroyl-, dimyristoyl-, diphalmitoyl-, or 1-palmitoyl-2-oleoylphosphatidic acid and optionally egg lecithin, dimyristoyl- or dipalmitoyllecithin, 1-palmitoyl-2-oleoyllecithin, beef brain phosphatidylserine or dipalmitoylphosphatidylserine are dispersed.
Claim 25
Dependent Claim
19 words
A process according to claim 1, wherein the aqueous phase is neutralised by addition of a physiologically acceptable base.
References Cited (Backward Citations)
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| 3.5 Years (Year 4) | 1989-10-27 | 1990-04-28 | 1990-10-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-10-27 | 1994-04-28 | 1994-10-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-10-27 | 1998-04-28 | 1998-10-28 | $7700.00 | Window Closed / Lapsed |