JP-2022530678-A5 -

Dates

Publication Date

20230510

Application Date

20200429

Description

In certain embodiments, the double-stranded nucleic acid inhibitor further comprises a 5'- phosphate mimetic at the 5' end of the sense strand and/or antisense strand. Modified Nucleoside: In this specification, the term “modified nucleoside” means a heterocyclic nitrogen base that is not bonded to a phosphate group or a modified phosphate group (as defined herein) and is N-glycosidically linked to a sugar (e.g., deoxyribose or ribose or an analog thereof) that contains one or more modified nucleic acid bases (as defined herein), universal nucleic acid bases (as defined herein), or modified sugar moieties (as defined herein). The above modified nucleic acid bases or universal nucleic acid bases (also called base analogs in this specification) generally refer to nucleic acid bases located at the 1' position of the nucleoside sugar moiety, other than adenine, guanine, cytosine, thymine, and uracil at the 1' position. In certain embodiments, the above modified nucleic acid bases or universal nucleic acid bases are nitrogen bases. In certain embodiments, the above modified nucleic acid bases or universal nucleic acid bases do not contain a nitrogen atom. See, for example, U.S. Patent Application Publication No. 20080274462. In certain embodiments, the above modified nucleotides do not contain a nucleic acid base (base-free). Suitable modified nucleic acid bases or universal nucleic acid bases or modified sugars in the context of this disclosure are described herein. In certain embodiments of the double-stranded nucleic acid inhibitor molecule, the sense strand has 21 to 25 nucleotides; the antisense strand has 22 nucleotides, including a single-stranded overhang of 6 to 10 nucleotides at the 3' end, and the 5' terminal nucleotide optionally contains a phosphate mimetic; the length of the first double helix is 12 to 16 base pairs; the second region of the sense strand contains a triloop, and the second double helix is 3 base pairs long and contains at least one Tm -increasing nucleotide. In certain embodiments, any nucleotide in the second double helix is a Tm -increasing nucleotide such as LNA. In certain embodiments of the double-stranded nucleic acid inhibitor molecule, the sense strand has 22 to 26 nucleotides; the antisense strand has 22 nucleotides including a single-stranded overhang of 6 to 10 nucleotides at its 3' end, and the 5' terminal nucleotide of the antisense strand optionally contains a phosphate mimetic; the length of the first double helix is 12 to 16 base pairs; the second region of the sense strand contains a tetraloop, and the second double helix is 3 base pairs long and contains at least one Tm -increasing nucleotide. In certain embodiments, any nucleotide in the second double helix is a Tm -increasing nucleotide such as LNA. In certain embodiments of the double-stranded nucleic acid inhibitor molecule, the sense strand has 21 to 31 nucleotides; the antisense strand has 22 nucleotides including a single-stranded overhang of 6 to 10 nucleotides at its 3' end, and the 5' terminal nucleotide of the antisense strand optionally contains a phosphate mimetic; the length of the first double helix is 12 to 16 base pairs; the second region of the sense strand contains a triloop, and the second double helix is 3 to 5 base pairs long and contains at least one Tm -increasing nucleotide, or 6 base pairs long and does not contain a Tm -increasing nucleotide. In certain embodiments, the second double helix is 3 base pairs long, and any nucleotide of the second double helix is a Tm -increasing nucleotide such as LNA. In certain embodiments of the double-stranded nucleic acid inhibitor molecule, the sense strand has 22 to 32 nucleotides; the antisense strand has 22 nucleotides including a single-stranded overhang of 6 to 10 nucleotides at its 3' end, and the 5' terminal nucleotide of the antisense strand optionally contains a phosphate mimetic; the length of the first double helix is 12 to 16 base pairs; the second region of the sense strand contains a tetraloop, and the second double helix is 3 to 5 base pairs long and contains at least one nucleotide that increases Tm , or is 6 base pairs long and does not contain a nucleotide that increases Tm . In certain embodiments, the second double helix is 3 base pairs long, and any nucleotide of the second double helix is a nucleotide that increases Tm , such as LNA. In certain embodiments, the double-stranded nucleic acid inhibitor molecule is conjugated to two to four sugar ligand moieties in the tetraloop or to two or three sugar ligand moieties in the triloop. In one embodiment, two nucleotides in the tetraloop are conjugated to the sugar ligand moiety. In one embodiment, three nucleotides in the tetraloop are conjugated to the sugar ligand moiety. In another embodiment, four nucleotides in the tetraloop are conjugated to the sugar ligand moiety. In one embodiment, two nucleotides in the triloop are conjugated to the sugar moiety. In one embodiment, three nucl