Lyophilizable and enhanced compacted nucleic acids
First Claim
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1. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single nucleic acid molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
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Abstract
Counterions of polycations used to compact nucleic acids profoundly affect shape and stability of particles formed. Shape is associated with differential serum nuclease resistance and colloidal stability. A surrogate for determining such properties that is easy to measure is the turbidity parameter. Shape also affects the suitability and efficacy of compacted nucleic acid complexes for transfecting cells by various routes into a mammalian body. Moreover, counterions such as acetate can protect compacted nucleic acid complexes from adverse effects of lyophilization
10 Citations
123 Claims
- 1. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single nucleic acid molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
- 8. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single nucleic acid molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, said polycation molecules having a nucleic acid binding moiety through which they are complexed to the nucleic acid, wherein said nucleic acid molecule encodes at least one functional protein, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
- 17. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single double-stranded cDNA molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, wherein said cDNA molecule encodes at least one functional protein, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
- 20. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single nucleic acid molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, wherein said nucleic acid molecule encodes at least one antisense nucleic acid, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
- 23. A non-naturally occurring composition comprising unaggregated nucleic acid complexes, each complex consisting essentially of a single nucleic acid molecule and one or more polycation molecules, wherein said complexes are formed by mixing said nucleic acid molecule and said polycation molecules, wherein prior to mixing said polycation molecules have a counterion selected from the group consisting of acetate, bicarbonate, and chloride, wherein said nucleic acid molecule is an RNA molecule, wherein said complexes are rod-shaped when visualized by transmission electron microscopy.
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26. Non-naturally occurring, soluble compacted complexes of a nucleic acid and a polycation molecule, wherein said complexes are rod-shaped when visualized by transmission electron microscopy, wherein each complex consists essentially of a single nucleic acid molecule and one or more polycation molecules, whereby said complexes are made by the process of:
mixing a nucleic acid with a polycation having acetate as a counterion, at a salt concentration sufficient for compaction of the complexes. - View Dependent Claims (30, 31, 106, 107, 108, 109)
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27. Non-naturally occurring, soluble compacted complexes of a nucleic acid and a polycation molecule, wherein the complexes are rod-shaped when visualized by transmission electron microscopy, wherein each complex consists essentially of a single nucleic acid molecule and one or more polycation molecules, whereby the complexes are made by the process of:
mixing a nucleic acid molecule with polycation molecules having a counterion selected from the group consisting of bicarbonate and chloride at a salt concentration sufficient for compaction of the complex, whereby unaggregated nucleic acid complexes are formed. - View Dependent Claims (32, 33, 110, 111, 112, 113)
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28. Non-naturally occurring, soluble compacted complexes of a nucleic acid and a polycation molecule, wherein the complexes are rod-shaped when visualized by transmission electron microscopy, wherein each complex consists essentially of a single nucleic acid molecule and one or more polycation molecules, whereby the complexes are made by the process of:
mixing a nucleic acid molecule with polycation molecules having acetate as a counterion in a solvent to form a complex, said mixing being performed in the absence of added salt, whereby the nucleic acid forms soluble complexes with the polycation molecules without forming aggregates. - View Dependent Claims (34, 35, 114, 115, 116, 117)
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29. Non-naturally occurring, soluble compacted complexes of a nucleic acid and a polycation wherein the complexes are rod-shaped when visualized by transmission electron microscopy, wherein each complex consists essentially of a single nucleic acid molecule and one or more polycation molecules, whereby the complexes are made by the process of:
mixing a nucleic acid molecule with polycation molecules having a counterion selected from the group consisting of bicarbonate and chloride in a solvent to form a complex, said mixing being performed in the absence of added salt, whereby the nucleic acid forms soluble complexes with the polycation molecules without forming aggregates. - View Dependent Claims (36, 37, 118, 119, 120, 121)
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Current AssigneeCopernicus Therapeutics, Inc.
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Original AssigneeCopernicus Therapeutics, Inc.
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InventorsCooper, Mark J., Kowalczyk, Tomasz H., Pasumarthy, Murali K., Costello, Maureen
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Granted Patent
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Time in Patent OfficeDays
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Field of Search
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US Class Current514/8
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CPC Class CodesA61K 48/00 Medicinal preparations cont...A61K 9/0043 NoseA61K 9/5146 obtained otherwise than by ...A61K 9/5192 ProcessesC12N 15/88 using microencapsulation, e...G01N 21/51 inside a container, e.g. in...
