Compounds and methods for modulating expression ApoB
View Patent ↗The present disclosure describes short antisense compounds, including such compounds comprising chemically-modified high-affinity monomers 8-16 monomers in length. Certain such short antisense compound are useful for the reduction of target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, provided herein are short antisense compounds comprising high-affinity nucleotide modifications useful for reducing a target RNA in vivo. Such short antisense compounds are effective at lower doses than previously described antisense compounds, allowing for a reduction in toxicity and cost of treatment. In addition, the described short antisense compounds have greater potential for oral dosing.
1. A method of modulating expression of an apolipoprotein B (ApoB) by contacting a nucleic acid encoding ApoB with a short antisense compound 10 to 14 monomers in length, comprising a 2′-deoxyribonucleotide gap region flanked on each side by at least one wing, wherein each wing independently comprises 1 to 3 high-affinity modified monomers, which are sugar-modified nucleotides that comprise a bridge between the 4′ and the 2′ position of the sugar; and wherein the short antisense compound is targeted to a nucleic acid encoding ApoB.
2. The method of claim 1 , wherein each of said high-affinity modified nucleotides confers a T m , of 1 to 4 degrees per nucleotide.
3. The method of claim 1 , wherein the conformation of each of said sugar-modified nucleotides is, independently, β-D or α-L.
4. The method of claim 1 , wherein each of said bridges independently comprises 1 or from 2 to 4 linked groups independently selected from —[C(R 1 )(R 2 )] n —, —C(R 1 )═C(R 2 )—, —C(R 1 )═N—, —C(═NR 1 )—, —C(═O)—, —C(═S)—, —O—, —Si(R 1 ) 2 —, —S(═O) x — and —N(R 1 )—;
wherein
x is 0, 1 or 2;
n is 1, 2, 3, or 4;
each R 1 and R 2 is, independently, H, a protecting group, hydroxyl, C 1 -C 12 alkyl, substituted C 1 -C 12 alkyl, C 2 -C 12 alkenyl, substituted C 2 -C 12 alkenyl, C 2 -C 12 alkynyl, substituted C 2 -C 12 alkynyl, C 5 -C 20 aryl, substituted C 5 -C 20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C 5 -C 7 alicyclic radical, substituted C 5 -C 7 alicyclic radical, halogen, OJ 1 , NJ 1 J 2 , SJ 1 , N 3 , COOJ 1 , acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O) 2 -J 1 ), or sulfoxyl (S(═O)-J 1 ); and
each J 1 and J 2 is, independently, H, C 1 -C 12 alkyl, substituted C 1 -C 12 alkyl, C 2 -C 12 alkenyl, substituted C 2 -C 12 alkenyl, C 2 -C 12 alkynyl, substituted C 2 -C 12 alkynyl, C 5 -C 20 aryl, substituted C 5 -C 20 aryl, acyl, (C(═O—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C 1 -C 12 aminoalkyl, substituted C 1 -C 12 aminoalkyl or a protecting group.
5. The method of claim 4 , wherein each of said bridges is, independently, 4′-CH 2 -2′, 4′-(CH 2 ) 2 -2′, 4′-CH 2 —O-2′, 4′-(CH 2 ) 2 —O-2′, 4′-CH 2 —O—N(R 1 )-2′ and 4′-CH 2 —N(R 1 )—O-2′-, wherein each R 1 is, independently, H, a protecting group or C 1 -C 12 alkyl.
6. The method of claim 1 , wherein at least one monomeric linkage is a modified monomeric linkage.
7. The method of claim 6 , wherein the modified monomeric linkage is a phosphorothioate linkage.
8. The method of claim 1 , wherein each monomeric linkage is a phosphorothioate internucleoside linkage.
9. The method of claim 1 , wherein the short antisense compound comprises a motif selected from 1-12-1; 2-10-2; 1-10-1; 1-10-2; 3-8-3; 2-8-2; 1-8-1; and 3-6-3; wherein, the first number represents the number of monomers in the 5′-wing, the second number represents the number of monomers in the gap, and the third number represents the number of monomers in the 3′ wing.
10. The method of claim 9 , wherein the motif is selected from 1-10-1; 2-10-2; and 1-9-2.
11. The method of claim 1 , wherein the short antisense compound comprises a motif selected from 1-1-10-2, 1-1-8-2, 1-1-6-3, and 1-2-8-2, wherein the first number represents the number of monomers in a first 5′ wing, the second number represents the number of monomers in a second 5′ wing, the third number represents the number of monomers in the gap, and the fourth number represents the number of monomers in the 3′ wing.
12. The method of claim 1 , wherein the short antisense compound comprises a motif selected from 2-10-1-1, 2-8-1-1, 3-6-1-1, and 2-8-2-1, wherein the first number represents the number of monomers in the 5′ wing, the second number represents the number of monomers in the gap, the third number represents the number of monomers in a first 3′ wing, and the fourth number represents the number of monomers in a second 3′ wing.
13. The method of claim 1 , wherein the short antisense compound comprises a motif selected from 1-1-8-1-1; 2-1-6-1-1; and 1-2-8-2-1, wherein the first number represents the number of monomers in a first 5′ wing, the second number represents the number of monomers in a second 5′ wing, the third number represents the number of monomers in the gap, the fourth number represents the number of monomers in a first 3′ wing and the fifth number represents the number of monomers in a second 3′ wing.
14. The method of claim 1 , wherein the ApoB nucleic acid is in a cell.
15. The method of claim 1 , wherein the ApoB nucleic acid is in an animal.
16. The method of claim 15 , wherein the animal is a human.
17. A method of inhibiting expression of ApoB RNA in an animal, comprising administering to said animal the short antisense compound 10 to 14 monomers in length, comprising a 2′-deoxyribonucleotide gap region flanked on each side by at least one wing, wherein each wing independently comprises 1 to 3 high-affinity modified monomers, which are sugar-modified nucleotides that comprise a bridge between the 4′ and the 2′ position of the sugar; and wherein the short antisense compound is targeted to a nucleic acid encoding ApoB.
18. A method of treating a cardiovascular disorder in an animal, comprising administering to an animal in need of such therapy a short antisense compound 10 to 14 monomers in length, comprising a 2′-deoxyribonucleotide gap region flanked on each side by at least one wing, wherein each wing independently comprises 1 to 3 high-affinity modified monomers, which are sugar-modified nucleotides that comprise a bridge between the 4′ and the 2′ position of the sugar; and wherein the short antisense compound is targeted to a nucleic acid encoding ApoB.
19. The method of claim 17 , wherein said administering results in decreased total serum cholesterol, serum LDL, serum VLDL, serum HDL, serum triglycerides, serum apolipoprotein(a) and/or free fatty acids in the animal.