Stable lipid-comprising drug delivery complexes and methods for their production
First Claim
1. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,wherein the polycation is a cationic peptide comprising polylysine, polyornithine, or polyarginine, or a polypeptide comprising polylysine, polyornithine, or polyarginine, andwherein the noncovalent complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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Abstract
Novel stable, concentrated, biologically active and ready-to-use lipid-comprising drug delivery complexes and methods for their production are described. The biological activity of the complexes produced are comparable to the formulations prepared according to the prior art admixture method and upon purification, the complexes produced by the method of this invention are 50 to 500 fold more concentrated than the complexes formed by admixture. The method described herein provides for the large scale production of lipid-comprising drug delivery systems useful for gene therapy and other applications.
165 Citations
36 Claims
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1. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide comprising polylysine, polyornithine, or polyarginine, or a polypeptide comprising polylysine, polyornithine, or polyarginine, and wherein the noncovalent complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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5. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a histone, or a fragment thereof, wherein the molecular weight of the polycation is between 300 and 200,000 Daltons, and wherein the complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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6. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a nuclear localization sequence, and wherein the complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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8. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a protamine sulfate, and wherein the complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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9. A noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is spermine or spermidine, and wherein the complex is formed by adding a nucleic acid to a solution comprising the at least one cationic lipid and the polycation.
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16. A method for delivering nucleic acid to cells comprising contacting the cells with a noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or a polypeptide comprising polylysine, polyornithine, or polyarginine, and wherein the noncovalent complex is formed by adding a nucleic acid to a purified solution comprising the at least one cationic lipid and the polycation.
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17. A method for delivering nucleic acid to cells comprising contacting the cells with a noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a histone, or a fragment thereof, wherein the molecular weight of the polycation is between 300 and 200,000 Daltons, and wherein the complex is formed by adding a nucleic acid to a purified solution comprising the at least one cationic lipid and the polycation.
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18. A method for delivering nucleic acid to cells comprising contacting the cells with a noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a nuclear localization sequence, and wherein the complex is formed by adding a nucleic acid to a purified solution comprising the at least one cationic lipid and the polycation.
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19. A method for delivering nucleic acid to cells comprising contacting the cells with a noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation is a cationic peptide or polypeptide comprising a protamine sulfate, and wherein the complex is formed by adding a nucleic acid to a purified solution comprising the at least one cationic lipid and the polycation.
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20. A method for delivering nucleic acid to cells comprising contacting the cells with a noncovalent nucleic acid/lipid/polycation complex having a net positive charge at pH 6.0-8.0, the complex comprising a nucleic acid, at least one cationic lipid and a polycation,
wherein the polycation comprises spermine or spermidine, and wherein the complex is formed by adding a nucleic acid to a purified solution comprising the at least one cationic lipid and the polycation.
Specification