Glucokinase (GCK) iRNA compositions and methods of use thereof
The invention relates to double stranded ribonucleic acid (dsRNA) compositions targeting a glucokinase (GCK) gene, as well as methods of inhibiting expression of a glucokinase (GCK) gene, and methods of treating subjects having a glycogen storage disease (GSD), e.g., type Ia GSD.
1. A double stranded ribonucleic acid (RNAi) agent for inhibiting expression of a glucokinase (GCK) gene,
wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein said antisense strand comprises at least 20 contiguous nucleotides differing by no more than 3 nucleotides from the complement of nucleotides 1466-1492 of the nucleotide sequence of SEQ ID NO:1,
wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification, and
wherein said sense strand is conjugated to a ligand attached at the 3′-terminus.
2. The double stranded RNAi agent of claim 1 , wherein at least one of the nucleotide modifications is selected from the group consisting of a 3′-terminal deoxy-thymine (dT) nucleotide modification, a 2′-O-methyl nucleotide modification, a 2′-fluoro nucleotide modification, a 2′-deoxy nucleotide modification, a locked nucleotide modification, an unlocked nucleotide modification, a conformationally restricted nucleotide modification, a constrained ethyl nucleotide modification, an abasic nucleotide modification, a 2′-amino nucleotide modification, a 2′-O-allyl-modified nucleotide modification, 2′-C-alkyl nucleotide modification, 2′-hydroxly nucleotide modification, a 2′-methoxyethyl nucleotide modification, a 2′-O-alkyl nucleotide modification, a morpholino nucleotide modification, a phosphoramidate nucleotide modification, a non-natural base comprising nucleotide modification, a tetrahydropyran nucleotide modification, a 1,5-anhydrohexitol nucleotide modification, a cyclohexenyl nucleotide modification, a nucleotide comprising a 5′-phosphorothioate group modification, a nucleotide comprising a 5′-methylphosphonate group modification, a nucleotide comprising a 5′ phosphate or 5′ phosphate mimic modification, a nucleotide comprising vinyl phosphate modification, a nucleotide comprising adenosine-glycol nucleic acid (GNA) modification, a nucleotide comprising thymidine-glycol nucleic acid (GNA)S-Isomer modification, a nucleotide comprising 2-hydroxymethyl-tetrahydrofurane-5-phosphate modification, a nucleotide comprising 2′-deoxythymidine-3′phosphate modification, a nucleotide comprising 2′-deoxyguanosine-3′-phosphate modification, and a terminal nucleotide linked to a cholesteryl derivative or a dodecanoic acid bisdecylamide group modification.
3. The double stranded RNAi agent of claim 1 , further comprising at least one phosphorothioate internucleotide linkage.
4. The double stranded RNAi agent of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide; or a 3′ overhang of at least 2 nucleotides.
5. The double stranded RNAi agent of claim 1 ,
wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
6. The double stranded RNAi agent of claim 5 , wherein the ligand is
7. A cell containing the double stranded RNAi agent of claim 1 .
8. A pharmaceutical composition for inhibiting expression of a glucokinase (GCK) gene comprising the double stranded RNAi agent of claim 1 .
9. A method of inhibiting expression of a glucokinase (GCK) gene in a cell, the method comprising:
(a) contacting the cell with the double stranded RNAi agent of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a GCK gene, thereby inhibiting expression of the GCK gene in the cell.
10. The method of claim 9 , wherein said cell is within a subject.
11. The method of claim 10 , wherein the subject is a human.
12. The method of claim 11 , wherein the human subject suffers from a disease or disorder that would benefit from reduction in GCK expression.
13. The method of claim 12 , wherein the disease or disorder is a glycogen storage disease (GSD).
14. A method of treating a subject having a disease or disorder that would benefit from reduction in expression of a glucokinase (GCK) gene, comprising administering to the subject a therapeutically effective amount of the double stranded RNAi agent of claim 1 , thereby treating said subject.
15. The method of claim 14 , wherein the disease or disorder is a glycogen storage disease (GSD).
16. The double stranded RNAi agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence 5′-UAUGAAGGUGAUCUCGCAGCUGG-3′ (SEQ ID NO:193).
17. The double stranded RNAi agent of claim 1 , wherein the antisense strand comprises the nucleotide sequence 5′ UAUGAAGGUGAUCUCGCAG-3′ (SEQ ID NO:194).
18. The double stranded RNAi agent of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-AGCUGCGAGAUCACCUUCAUA-3′ (SEQ ID NO:104) and the antisense strand comprises the nucleotide sequence 5′-UAUGAAGGUGAUCUCGCAGCUGG-3′ (SEQ ID NO:193).
19. The double stranded RNAi agent of claim 1 , wherein the sense strand comprises the nucleotide sequence 5′-CUGCGAGAUCACCUUCAUA-3′ (SEQ ID NO:105) and the antisense strand comprises the nucleotide sequence 5′-UAUGAAGGUGAUCUCGCAG-3′ (SEQ ID NO:194).
20. The double stranded RNAi agent of claim 1 , wherein each strand is independently 20-30 nucleotides in length.
21. The double stranded RNAi agent of claim 1 , wherein each strand is independently 20-25 nucleotides in length.
22. The double stranded RNAi agent of claim 1 , wherein each of the sense strand and the antisense strand independently is 21 to 23 nucleotides in length.
23. The double stranded RNAi agent of claim 1 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.
24. The double stranded RNAi agent of claim 1 , wherein at least one of the nucleotide modifications is selected from the group consisting of a 2′-O-methyl nucleotide modification and a 2′fluoro nucleotide modification.
25. The method of claim 14 , further comprising administering a sodium-glucose co-transporter 2 (SGLT2) inhibitor to the subject.
26. A method of inhibiting the expression of a glucokinase (GCK) gene in a subject, the method comprising administering to said subject a therapeutically effective amount of the double stranded RNAi agent of claim 1 , thereby inhibiting the expression of GCK in said subject.