COMPOSITIONS AND METHODS OF USING PRKAG2-TARGETING ANTIBODY-OLIGONUCLEOTIDE CONJUGATES
Disclosed herein are polynucleic acid molecules, pharmaceutical compositions, and methods of use for PRKAG2 antibody oligonucleotide conjugates (AOC).
1 . A polynucleotide conjugate comprising an anti-transferrin receptor antibody or antigen-binding fragment thereof conjugated to a polynucleotide, wherein the polynucleotide is at least 90% complementary to a target sequence of a human PRKAG2 mRNA and mediates downregulation of the human PRKAG2 mRNA in a cardiac muscle cell, wherein the target sequence corresponds to positions 1897-1917 or positions 1927-1947 of the human PRKAG2 mRNA (SEQ ID NO: 237) from the 5′ end.
2 . The polynucleotide conjugate of claim 1 , wherein the human PRKAG2 mRNA comprises a gain of function mutation.
3 . The polynucleotide conjugate of claim 1 , wherein the polynucleotide is a single-stranded antisense oligonucleotide (ASO) or a double-stranded small interfering RNA (siRNA) comprising a guide strand and a passenger strand.
4 . The polynucleotide conjugate of claim 3 , wherein the guide strand comprises a nucleic acid sequence that is at least 95% complementary to the target sequence.
5 . The polynucleotide conjugate of claim 4 , wherein the passenger strand comprises a nucleic acid sequence that is at least 95% complementary to the guide strand.
6 . The polynucleotide conjugate of claim 1 , wherein the polynucleotide comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety.
7 . The polynucleotide conjugate of claim 6 , wherein the at least one 2′ modified nucleotide comprises:
2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O-N-methylacetamido (2′-O-NMA) modified nucleotide, a locked nucleic acid (LNA), an ethylene nucleic acid (ENA), or a combination thereof.
8 . The polynucleotide conjugate of claim 6 , wherein the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage.
9 . The polynucleotide conjugate of claim 1 , wherein the polynucleotide comprises a 5′-terminal vinylphosphonate modified nucleotide.
10 . The polynucleotide conjugate of claim 1 , wherein the guide strand comprises the nucleic acid sequence of SEQ ID NOs: 211 or 212 and the passenger strand comprises the nucleic acid sequence of SEQ ID NOs: 223 or 224.
11 . The polynucleotide conjugate of claim 1 , wherein the polynucleotide conjugate has a polynucleotide to antibody ratio of from about 1 to about 4.
12 . The polynucleotide conjugate of claim 1 , wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a non-human antibody or antigen binding fragment thereof, a human antibody or antigen binding fragment thereof, a humanized antibody or antigen binding fragment thereof, a chimeric antibody or antigen binding fragment thereof, a monoclonal antibody or antigen binding fragment thereof, a monovalent Fab′, a divalent Fab2, a single-chain variable fragment (scFv), a diabody, a minibody, a nanobody, a single-domain antibody (sdAb), or a camelid antibody or antigen binding fragment thereof.
13 . The polynucleotide conjugate of claim 1 , wherein the anti-transferrin receptor antibody or antigen-binding fragment thereof comprises the VH sequence of SEQ ID NOs: 256, 257, 258, 259, or 260, and the VL sequence of SEQ ID NOs: 261, 262, 263, 264, or 265.
14 . The polynucleotide conjugate of claim 1 , wherein the polynucleotide conjugate comprises a linker connecting the anti-transferrin receptor antibody or antigen-binding fragment thereof to the polynucleotide.
15 . The polynucleotide conjugate of claim 14 , wherein the linker comprises a maleimide group.
16 . A polynucleotide molecule for modulating human PRKAG2 mRNA expression, wherein the polynucleotide molecule is a double-stranded siRNA comprising a guide strand and a passenger strand, wherein the guide strand comprises a nucleic acid sequence that is at least 95% complementary to a nucleic acid sequence corresponding to positions 1897-1917 or positions 1927-1947 of the human PRKAG2 mRNA (SEQ ID NO: 237) from the 5′ end.
17 . The polynucleotide molecule of claim 16 , wherein the passenger strand comprises a nucleic acid sequence that is at least 95% complementary to the guide strand.
18 . The polynucleotide molecule of claim 16 , wherein the polynucleotide molecule comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety.
19 . The polynucleotide molecule of claim 18 , wherein the at least one 2′ modified nucleotide comprises:
2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O-N-methylacetamido (2′-O-NMA) modified nucleotide, a locked nucleic acid (LNA), an ethylene nucleic acid (ENA), or a combination thereof.
20 . The polynucleotide molecule of claim 18 , wherein the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage.
21 . The polynucleotide molecule of claim 16 , wherein the polynucleotide molecule comprises a 5′-terminal vinylphosphonate modified nucleotide.
22 . The polynucleotide molecule of claim 16 , wherein the guide strand comprises the nucleic acid sequence of SEQ ID NOs: 211 or 212 and the passenger strand comprises the nucleic acid sequence of SEQ ID NOs: 223 or 224.
23 . A method of treating cardiomyopathy in a subject in need thereof comprising administering to said subject a polynucleotide conjugate comprising an anti-transferrin receptor antibody or antigen-binding fragment thereof conjugated to a polynucleotide, wherein the polynucleotide is at least 90% complementary to a target sequence of a human PRKAG2 mRNA, thereby treating cardiomyopathy in said subject, wherein the target sequence corresponds to positions 1897-1917 or positions 1927-1947 of the human PRKAG2 mRNA (SEQ ID NO: 237) from the 5′ end.
24 . The method of claim 23 , wherein the cardiomyopathy is caused by at least one of: a glycogen storage disease, PRKAG2 syndrome or PRKAG2 cardiac syndrome.
25 . The method of claim 24 , wherein the PRKAG2 syndrome or the PRKAG2 cardiac syndrome is caused by a mutated human PRKAG2 mRNA that has a gain of function.