US6071533A
Expired / Lapsed
US
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Preparation of stable formulations of lipid-nucleic acid complexes for efficient in vivo delivery
Assignee: University of California
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Filing: 1997-11-10
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Granted: 2000-06-06
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Expires: 2017-11-10
Abstract
Abstract The present invention provides for lipid:nucleic acid complexes that have increased shelf life and high transfection activity in vivo following intravenous injection, and methods of preparing such complexes. The methods generally involve contacting a nucleic acid with an organic polycation to produce a condensed nucleic acid, and then combining the condensed nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce the lipid:nucleic acid complex. This complex can be further stabilized by the addition of a hydrophilic polymer attached to hydrophobic side chains. The complex can also be made specific for specific cells, by incorporating a targeting moiety such as an Fab' fragment attached to a hydrophilic polymer.
Inventors
- 👤 Demetrios Papahadjopoulos
- 👤 Keelung Hong
- 👤 Weiwen Zheng
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 11:05
CPC Classifications
C; C07; C07H; C07H21/00; A; A61; A61K; A61K47/00; A61K47/06; A61K47/08; A61K47/10; A61K47/50; A61K47/51; A61K47/54; A61K47/542; A61K47/543; A61K47/68; A61K47/6835; A61K47/6849; A61K47/69; A61K47/6905; A61K47/6911; A61K47/6913; A61K48/00; A61K9/00; A61K9/0012; A61K9/0019; A61K9/10; A61K9/127; A61K9/1271; A61K9/1272
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
86 words
A method of increasing the shelf life of a lipid:nucleic acid complex, said method comprising the steps of: combining a nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce said lipid:nucleic acid complex; and mixing said lipid:nucleic acid complex with a hydrophilic polymer in a molar ratio from about 0.1 to about 10% hydrophilic polvmer to lipid within the lipid:nucleic acid complex, wherein said lipid:nucleic acid complex has increased shelf life as compared to an identical lipid:nucleic acid complex lacking said hydrophilic polymer.
Claim 2
Independent Claim
86 words
A method of increasing the shelf life of a lipid:nucleic acid complex, said method comprising the steps of: combining a nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce said lipid:nucleic acid complex; and mixing said lipid:nucleic acid complex with a hydrophilic polymer in a molar ratio from about 0.1 to about 10% hydrophilic polvmer to lipid within the lipid:nucleic acid complex, wherein said lipid:nucleic acid complex has increased shelf life as compared to an identical lipid:nucleic acid complex lacking said hydrophilic polymer.
Claim 3
Dependent Claim
42 words
The method of claim 1, wherein said hydrophilic polymer is selected from the group consisting of polyethylene glycol (PEG), polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE), polyethylene glycol derivatized with tween, polyethylene glycol derivatized with distearoylphosphatidylethanolamine (PEG-DSPE), and ganglioside G M1 .
Claim 4
Dependent Claim
19 words
The method of claim 1, wherein said nucleic acid is selected from the group consisting of DNA and RNA.
Claim 5
Dependent Claim
11 words
The method of claim 1, wherein said nucleic acid is DNA.
Claim 6
Dependent Claim
26 words
The method of claim 1, wherein said step of combining said nucleic acid with said lipid comprises first forming a liposome comprising said amphiphilic cationic lipid.
Claim 7
Dependent Claim
38 words
The method of claim 1, wherein said step of combining said nucleic acid with said lipid comprises combining said lipid and said nucleic acid in a ratio ranging from about 1 to about 20 nmole lipid:μg nucleic acid.
Claim 8
Dependent Claim
74 words
The method of claim 1, wherein said method comprises the steps of: combining an expression cassette with said lipid comprising said amphiphilic cationic lipid to produce a lipid:expression cassette complex; and mixing said lipid:expression cassette complex with polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE), wherein said lipid:expression cassette complex has increased shelf life at about 4° C. as compared to an identical lipid:expression cassette complex lacking said polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE).
Claim 9
Dependent Claim
19 words
The method of claim 7, wherein said lipid comprises polyethylene glycol attached to a Fab' fragment of an antibody.
Claim 10
Dependent Claim
12 words
The method of claim 1, wherein said lipid:nucleic acid complex is lyophilized.
Claim 11
Independent Claim
90 words
A method of increasing the shelf life of a lipid:nucleic acid complex, said method comprising the steps of: combining a nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce said lipid:nucleic acid complex, wherein said lipid and said nucleic acid are combined in a ratio ranging from about 1 to about 20 nmole lipid:μg nucleic acid; and mixing said lipid:nucleic acid complex with a hydrophilic polymer, wherein said lipid:nucleic acid complex has increased shelf life as compared to an identical lipid:nucleic acid complex lacking said hydrophilic polymer.
Claim 12
Independent Claim
90 words
A method of increasing the shelf life of a lipid:nucleic acid complex, said method comprising the steps of: combining a nucleic acid with a lipid comprising an amphiphilic cationic lipid to produce said lipid:nucleic acid complex, wherein said lipid and said nucleic acid are combined in a ratio ranging from about 1 to about 20 nmole lipid:μg nucleic acid; and mixing said lipid:nucleic acid complex with a hydrophilic polymer, wherein said lipid:nucleic acid complex has increased shelf life as compared to an identical lipid:nucleic acid complex lacking said hydrophilic polymer.
Claim 13
Dependent Claim
42 words
The method of claim 10, wherein said hydrophilic polymer is selected from the group consisting of polyethylene glycol (PEG), polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE), polyethylene glycol derivatized with tween, polyethylene glycol derivatized with distearoylphosphatidylethanolamine (PEG-DSPE), and ganglioside G M1 .
Claim 14
Dependent Claim
19 words
The method of claim 10, wherein said nucleic acid is selected from the group consisting of DNA and RNA.
Claim 15
Dependent Claim
11 words
The method of claim 10, wherein said nucleic acid is DNA.
Claim 16
Dependent Claim
26 words
The method of claim 10, wherein said step of combining said nucleic acid with said lipid comprises first forming a liposome comprising said amphiphilic cationic lipid.
Claim 17
Dependent Claim
47 words
The method of claim 10, wherein said step of mixing said lipid:nucleic acid complex with said hydrophilic polymer comprises mixing said hydrophilic polymer and said lipid:nucleic acid complex in a molar ratio from about 0.1 to about 10% hydrophilic polymer to lipid within the lipid:nucleic acid complex.
Claim 18
Dependent Claim
74 words
The method of claim 10, wherein said method comprises the steps of: combining an expression cassette with said lipid comprising said amphiphilic cationic lipid to produce a lipid:expression cassette complex; and mixing said lipid:expression cassette complex with polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE), wherein said lipid:expression cassette complex has increased shelf life at about 4° C. as compared to an identical lipid:expression cassette complex lacking said polyethylene glycol derivatized with phosphatidyl ethanolamine (PEG-PE).
Claim 19
Dependent Claim
19 words
The method of claim 10, wherein said lipid comprises polyethylene glycol attached to a Fab' fragment of an antibody.
Claim 20
Dependent Claim
12 words
The method of claim 10, wherein said lipid:nucleic acid complex is lyophilized.
References Cited (Backward Citations)
US5783210A
Cited
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