RNAI constructs for inhibiting PNPLA3 expression and methods of use thereof
The present invention relates to RNAi constructs for reducing expression of the PNPLA3 gene. Methods of using such RNAi constructs to treat or prevent liver disease, nonalcoholic fatty liver disease (NAFLD) are also described.
1 . An RNAi construct comprising a sense strand and an antisense strand, wherein the antisense strand comprises an antisense sequence comprising asGfscca[Ab]UfguagAfaAfggcaugasusu (SEQ ID NO: 3004),
wherein (i) a, u, g, and c are 2′-O-methyl ribonucleotides, (ii) [Ab] indicates an abasic nucleotide, (iii) Af, Uf, and Gf are 2′-deoxy-2′-fluoro ribonucleotides, and (iv) “s” indicates a phosphorothioate internucleotide linkage, and wherein the RNAi construct inhibits the expression of Patatin-Like Phospholipase Domain Containing 3 (PNPLA3).
2 . The RNAi construct of claim 1 , wherein the sense strand comprises a sense sequence comprising
{sGalNAc3K2AhxC6}ucaugccuUfuCfUfAfCf[LNA-A]guggcs{invAb}(SEQ ID NO: 3005);
wherein (i) a, u, g, and c are 2′-O-methyl ribonucleotides, (ii) {invAb} indicates an inverted abasic nucleotide, (iii) Af, Uf, and Gf are 2′-deoxy-2′-fluoro ribonucleotides, (iv) “s” indicates a phosphorothioate internucleotide linkage, (v) GalNAc3K2AhxC6 is an N-acetyl-galactosamine-containing ligand; and (vi) LNA indicates a locked nucleic acid.
3 . The RNAi construct of claim 1 , wherein the RNAi construct comprises at least one blunt end.
4 . The RNAi construct of claim 1 , wherein the RNAi construct comprises at least one nucleotide overhang of 1 to 4 unpaired nucleotides.
5 . The RNAi construct of claim 4 , wherein the nucleotide overhang has 2 unpaired nucleotides.
6 . The RNAi construct of claim 1 , wherein the RNAi construct reduces the expression level of PNPLA3 in liver cells following incubation with the RNAi construct as compared to the PNPLA3 expression level in liver cells that have been incubated with a control RNAi construct.
7 . A pharmaceutical composition comprising the RNAi construct of claim 1 and a pharmaceutically acceptable carrier, excipient, or diluent.
8 . A method for reducing the expression of PNPLA3 in a patient in need thereof comprising administering to the patient the RNAi construct of claim 1 .
9 . An RNAi construct comprising a sense sequence comprising {sGalNAc3K2AhxC6}ucaugccuUfuCfUfAfCfa[sLNA-G]uggcs{invAb}(SEQ ID NO: 3003) and antisense sequence comprising asGfscca[Ab]UfguagAfaAfggcaugasusu (SEQ ID NO: 3004): wherein (i) a, u, g, and c are 2′-O-methyl ribonucleotides, (ii) {invAb} indicates an inverted abasic nucleotide, (iii) Af, Uf, and Gf are 2′-deoxy-2′-fluoro ribonucleotides, (iv) “s” indicates a phosphorothioate internucleotide linkage, (v) GalNAc3K2AhxC6 is an N-acetyl-galactosamine-containing ligand; and (vi) LNA indicates a locked nucleic acid.
10 . A pharmaceutical composition comprising the RNAi construct of claim 9 and a pharmaceutically acceptable carrier, excipient, or diluent.
11 . A method for reducing the expression of PNPLA3 in a patient in need thereof comprising administering to the patient the RNAi construct of claim 9 .
12 . A pharmaceutical composition comprising the RNAi construct of claim 2 and a pharmaceutically acceptable carrier, excipient, or diluent.
13 . A method for reducing the expression of PNPLA3 in a patient in need thereof comprising administering to the patient the RNAi construct of claim 2 .