Three component chimeric antisense oligonucleotides
First Claim
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1. A chimeric antisense oligonucleotide comprising:
- a 5'"'"' terminus;
a 3'"'"' terminus and from 11 to 59 5'"'"'→
3'"'"'-linked nucleotides capable of contiguously hybridizing to a specific RNA and independently selected from the group consisting of 2'"'"'-modified phosphodiester nucleotides, and 2'"'"'-modified P-alkyloxyphosphotriester nucleotides; and
wherein said 5'"'"' terminal nucleoside is attached to an RNAse H-activating region capable of contiguously hybridizing to the specific RNA and of between three and ten contiguous phosphorothioate-linked deoxyribonucleotides, and wherein the 3'"'"' terminus of said oligonucleotide is drawn from the group consisting of;
an inverted deoxyribonucleotide, a contiguous stretch of one to three phosphorothioate 2'"'"'-modified ribonucleotides, a biotin group, and a P-alkyloxyphosphodiester nucleotide.
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Abstract
This invention relates to antisense oligonucleotides that target mRNAs in cells as substrates for the cellular enzyme RNase H and thereby cause specific degradation of the targeted mRNA. The oligonucleotides have three components: a RNase H activating region, a complementarity region and 3'"'"' and 5'"'"' ends. The invention optimizes each of the components to resist intracellular nucleases, to increase hybridization to target mRNA, to specifically inactivate target mRNA in cells, and to decrease cytotoxicity.
173 Citations
23 Claims
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1. A chimeric antisense oligonucleotide comprising:
- a 5'"'"' terminus;
a 3'"'"' terminus and from 11 to 59 5'"'"'→
3'"'"'-linked nucleotides capable of contiguously hybridizing to a specific RNA and independently selected from the group consisting of 2'"'"'-modified phosphodiester nucleotides, and 2'"'"'-modified P-alkyloxyphosphotriester nucleotides; and
wherein said 5'"'"' terminal nucleoside is attached to an RNAse H-activating region capable of contiguously hybridizing to the specific RNA and of between three and ten contiguous phosphorothioate-linked deoxyribonucleotides, and wherein the 3'"'"' terminus of said oligonucleotide is drawn from the group consisting of;
an inverted deoxyribonucleotide, a contiguous stretch of one to three phosphorothioate 2'"'"'-modified ribonucleotides, a biotin group, and a P-alkyloxyphosphodiester nucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19)
- a 5'"'"' terminus;
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17. A chimeric antisense oligonucleotide, comprising:
- a 5'"'"' terminus;
a 3'"'"' terminus; and
from 11 to 59 5'"'"'→
3'"'"'-linked nucleotides capable of contiguously hybridizing to a specific RNA and independently selected from the group consisting of 2'"'"'-modified phosphodiester nucleotides, and 2'"'"'-modified P-alkyloxyphosphotriester nucleotides; and
wherein said 3'"'"' terminal nucleoside is attached to an RNAse H-activating region capable of contiguously hybridizing to the specific RNA and of between three and ten contiguous phosphorothioate-linked deoxyribonucleotides, and wherein the 5'"'"' terminus of said oligonucleotide is drawn from the group consisting of;
an inverted deoxyribonucleotide, a contiguous stretch of one to three phosphorothioate 2'"'"'-modified ribonucleotides, a biotin group, and a P-alkyloxyphosphodiester nucleotide.
- a 5'"'"' terminus;
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20. A method of specifically cleaving an RNA in a cell containing RNAse H which comprises administering an effective amount of an oligonucleotide complementary to the RNA comprising:
- a 5'"'"' terminus;
a 3'"'"' terminus; and
from 11 to 59 5'"'"'→
3'"'"'-linked nucleotides capable of contiguously hybridizing to the RNA and independently selected from the group consisting of 2'"'"'-modified phosphodiester nucleotides, 2'"'"'-modified P-alkyloxyphosphotriester nucleotides; and
wherein said 5'"'"' terminal nucleoside is attached to an RNAse H-activating region capable of contiguously hybridizing to the RNA and of between three and ten contiguous phosphorothioate-linked deoxyribonucleotides, and wherein the 3'"'"' terminus of said oligonucleotide is drawn from the group consisting of;
an inverted deoxyribonucleotide, a contiguous stretch of one to three phosphorothioate deoxyribonucleotides, phosphorothioate 2'"'"'-modified ribonucleotides, a biotin group, and a P-alkyloxyphosphodiester-linked nucleotide.
- a 5'"'"' terminus;
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21. A chimeric antisense oligonucleotide comprising:
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a) a 5'"'"' terminal RNAse H activation region capable of contiguously hybridizing to a specific RNA and having between 5 and 10 contiguous deoxyphosphorothioate nucleotides; b) between 11 to 59 contiguous 5'"'"'→
3'"'"'-linked 2'"'"'-methoxy ribonucleotides capable of contiguously hybridizing to the specific RNA; andc) an exonuclease blocking group present at the 3'"'"' end of the oligonucleotide that is drawn from the group consisting of;
a non-5'"'"'-3'"'"' phosphodiester-linked nucleotide, from one to three contiguous 5'"'"'-3'"'"'-linked modified nucleotides, and a non-nucleotide chemical blocking group. - View Dependent Claims (22, 23)
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Specification