Methods for the Delivery of Oligomeric Compounds
First Claim
1. A method of inhibiting the expression of a gene in a cell, the method comprising administering to the cell a composition comprising a hybridization complex, wherein the hybridization complex comprises:
- (a) a first oligonucleotide strand, wherein the first oligonucleotide strand comprises a fully or partially modified oligonucleotide that hybridizes to at least a portion of an mRNA molecule transcribed from the gene; and
(b) a second oligonucleotide strand, wherein the second oligonucleotide strand comprises a deoxyribonucleic acid that is complementary to at least a portion of the first strand, whereby the first oligonucleotide strand and the second oligonucleotide strand enter the cell and the expression of the gene in the cell is inhibited.
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Accused Products
Abstract
The presently disclosed subject matter relates to the delivery of oligonucleotides to cells through the delivery of a composition or reagent comprising a hybridization complex comprising a first antisense oligonucleotide which is modified to have a higher stability against degradation, and a second sense oligonucleotide which is prone to degradation. The presently disclosed subject matter furthermore relates to dendrimeric bioconjugates and compositions or reagents comprising them, wherein the bioconjugate comprises a conjugate moiety coupled to a dendrimeric structure and to their use to deliver oligomeric compounds including oligonucleotides or duplexes, as described above, to cells for modulation of gene expression (i.e. antisense or antigene therapy/research, RNA interference).
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Citations
44 Claims
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1. A method of inhibiting the expression of a gene in a cell, the method comprising administering to the cell a composition comprising a hybridization complex, wherein the hybridization complex comprises:
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(a) a first oligonucleotide strand, wherein the first oligonucleotide strand comprises a fully or partially modified oligonucleotide that hybridizes to at least a portion of an mRNA molecule transcribed from the gene; and (b) a second oligonucleotide strand, wherein the second oligonucleotide strand comprises a deoxyribonucleic acid that is complementary to at least a portion of the first strand, whereby the first oligonucleotide strand and the second oligonucleotide strand enter the cell and the expression of the gene in the cell is inhibited. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of enhancing the uptake of a single-stranded oligonucleotide by a cell, the method comprising:
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(a) hybridizing to the single-stranded oligonucleotide a deoxyribonucleic acid molecule that is complementary to a subsequence of the single-stranded oligonucleotide to create a double-stranded molecule; and (b) contacting the cell with the double-stranded molecule, whereby uptake of the single-stranded oligonucleotide by the cell is enhanced. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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- 34. A composition comprising a dendrimeric bioconjugate and an oligomeric compound, wherein the dendrimeric bioconjugate comprises a conjugate moiety coupled to a dendrimeric structure.
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39. A method of enhancing the uptake of an oligomeric compound, the method comprising:
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(a) combining the oligomeric compound with a dendrimeric bioconjugate in a composition, wherein the dendrimeric bioconjugate comprises a conjugate moiety coupled to a dendrimeric structure; and (b) contacting the cell with the composition.
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- 40. A method of inhibiting the expression of a gene in a cell, the method comprising administering to the cell a composition comprising an oligomeric compound and a dendrimeric bioconjugate, wherein the dendrimeric bioconjugate comprises a conjugate moiety coupled to a dendrimeric structure.
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43. A method for the preparation of cholesterol-dendrimeric oligonucleotide bioconjugate, the method comprising:
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(a) derivatizing cholesterol to obtain a free carboxylgroup; (b) coupling to the derivatized cholesterol Fmoc-lysine-OMe or an analogous structure comprising a protected amino group and a protected carboxylgroup which are separately cleavable; (c) deprotecting the carboxyl group; (d) coupling the obtained compound to a solid phase through an acid labile function, like an ester; (e) deprotecting the amino group; (f) repetitive steps of [1] coupling of a compound with two protected amino functions and a free carboxylgroup like (Fmoc)2-Lys-OH; and [2] deprotecting the amino-funtion protecting groups according to the amount of dendrimeric units requested; (g) coupling a linker with a free hydroxyl as reactive group; and (h) performing standard oligonucleotide synthesis followed by cleaving the compound of from the solid phase.
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44. A method of enhancing the uptake in a cell of an oligomeric compound mediating RNA interference such as siRNAs, the method comprising combining the oligomeric compound with a dendrimeric structure such as a cationic polymer.
Specification