RNA-EDITING OLIGONUCLEOTIDES AND USES THEREOF
The present disclosure features useful compositions and methods to treat disorders for which deamination of an adenosine in an mRNA produces a therapeutic result, e.g., in a subject in need thereof.
1 . An oligonucleotide comprising the structure:
[A m ]-X 1 -X 2 -X 3 —[B n ]
wherein each of A and B is a nucleotide;
m and n are each, independently, an integer from 1 to 50;
X 1 , X 2 , and X 3 are each, independently, a nucleotide, wherein at least one of X 1 , X 2 , or X 3 has the structure of any one of Formula I-V:
wherein N 1 is hydrogen or a nucleobase;
R 1 is hydroxy, halogen, or C 1 -C 6 alkoxy;
R 2 is hydrogen, hydroxy, halogen, or C 1 -C 6 alkoxy;
R 3 is hydrogen, hydroxy, halogen, or C 1 -C 6 alkoxy;
R 4 is hydrogen, hydroxy, halogen, or C 1 -C 6 alkoxy; and
R 5 is hydrogen, hydroxy, halogen, or C 1 -C 6 alkoxy.
2 . The oligonucleotide of claim 1 , wherein R 4 is hydrogen and R 5 is not hydrogen or hydroxy, R 5 is hydrogen and R 4 is not hydrogen, or R 5 is hydroxy and R 4 is not hydrogen.
3 . The oligonucleotide of claim 1 , wherein at least 80% of the nucleotides of [A m ] and/or [B n ] include a nucleobase, a sugar, and an internucleoside linkage.
4 . The oligonucleotide of claim 1 , wherein R 1 is hydroxy, halogen, or OCH 3 .
5 . The oligonucleotide of claim 1 , wherein R 2 is hydrogen.
6 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , or X 3 has the structure of Formula I, Formula II, or Formula V; and none of X 1 , X 2 , or X 3 has the structure of Formula IV or Formula III.
7 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , or X 3 has the structure of Formula I or Formula II; and none of X 1 , X 2 , or X 3 has the structure of Formula III, Formula IV, or Formula V.
8 . The oligonucleotide of claim 1 , wherein the halogen is fluoro.
9 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , and X 3 has the structure of Formula I, wherein R 1 is fluoro and N 1 is a nucleobase.
10 - 12 . (canceled)
13 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , and X 3 has the structure of Formula I, wherein R 1 is hydroxy and N 1 is a nucleobase.
14 - 16 . (canceled)
17 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , and X 3 has the structure of Formula I, wherein R 1 is methoxy and N 1 is a nucleobase.
18 - 20 . (canceled)
21 . The oligonucleotide of claim 1 , wherein at least one of X 1 , X 2 , and X 3 has the structure of Formula II, wherein R 2 is hydrogen and N 1 is a nucleobase.
22 . (canceled)
23 . The oligonucleotide of claim 1 , wherein at least one of X 1 and X 2 has the structure of Formula V.
24 . (canceled)
25 - 59 . (canceled)
60 . The oligonucleotide of claim 1 , wherein at least 20% of the nucleotides of [A m ] and [B n ] combined are 2′-O-methyl-nucleotides.
61 - 63 . (canceled)
64 . The oligonucleotide of claim 1 , wherein A and B combined consist of 18 to 80 nucleotides.
65 - 66 . (canceled)
67 . The oligonucleotide of claim 1 , wherein m and n are each, independently, an integer from 5 to 40; at least one of X 1 , X 2 , and X 3 has the structure of Formula I, wherein R 1 is fluoro, hydroxy, or methoxy and N 1 is a nucleobase, or the structure of Formula V, wherein R 4 is hydrogen and R 5 is hydrogen; each of X 1 , X 2 , and X 3 that does not have the structure of Formula I or Formula V is a ribonucleotide; [A m ] and [B n ] each comprise at least five terminal 2′-O-methyl-nucleotides and at least four terminal phosphorothioate linkages; and at least 20% of the nucleotides of [A m ] and [B n ] combined are 2′-O-methyl-nucleotides.
68 . The oligonucleotide of claim 1 , wherein the oligonucleotide further comprises one or more adenosine deaminase acting on RNA (ADAR)-recruiting domains.
69 . A conjugate comprising an oligonucleotide of claim 1 conjugated to a targeting moiety.
70 . (canceled)
71 . A complex comprising:
an oligonucleotide of claim 1 ; and
an mRNA,
wherein the oligonucleotide or conjugate and mRNA are hybridized to each other and the complex comprises a first mismatch at an adenosine of the mRNA.
72 - 79 . (canceled)
80 . A method of producing a complex, the method comprising contacting a cell with an oligonucleotide of claim 1 .
81 . A method for deamination of an adenosine in an mRNA, the method comprising contacting a cell with an oligonucleotide of claim 1 .
82 . A method of treating a disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of an oligonucleotide of claim 1 .
83 . (canceled)