POLYNUCLEOTIDE AGENTS TARGETING PROGRAMMED CELL DEATH 1 LIGAND 1 (PD-L1) AND METHODS OF USE THEREOF
The invention relates to polynucleotide agents targeting programmed cell death 1 ligand 1 (PD-L1) gene, and methods of using such polynucleotide agents to inhibit expression of PD-L1 and to treat subjects having a PD-L1-associated disorder.
1 . An antisense polynucleotide agent for inhibiting expression of a Programmed cell death 1 ligand 1 (PD-L1) gene, wherein the agent comprises 4 to 50 contiguous nucleotides, wherein at least one of the contiguous nucleotides comprises a nucleotide modification, and wherein the nucleotide sequence of the agent is 80% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of SEQ ID NOs:1-5.
2 . The antisense polynucleotide agent of claim 1 , wherein the agent comprises 4 to 50 contiguous nucleotides, wherein at least one of the contiguous nucleotides comprises a nucleotide modification, and wherein the nucleotide sequence of the agent is 80% complementary over its entire length to the equivalent region of the nucleotide sequence of any one of residues 10-29; 44-75; 44-141; 78-141; 187-305; 309-349; 351-467; 309-467; 472-503; 505-570; 472-570; 571-590; 505-590; 606-647; 681-755; 769-788; 793-811; 815-834; 859-911; 1000-1019; 1044-1076; 1100-1152; 1177-1196; 1211-1241; 1242-1261; 1211-1261; 1277-1307; 1309-1328; 1277-1328; 1342-1373; 1374-1407; 1418-1450; 1462-1493; 1497-1582; 1650-1713; 1650-1756; 1716-1756; 1781-1879; 1915-1945; 1958-1977; 1990-2009; 2112-2131; 2167-2186; 2211-2230; 2288-2307; 2332-2351; 2364-2383; 2552-2571; 2575-2594; 2552-2594; 2652-2671; 2739-2801; 2826-2878; 2904-2933; 2970-2989; 3013-3032; 3035-3054; 3113-3142; 3158-3199 of SEQ ID NO:1
3 .- 7 . (canceled)
8 . The agent of claim 1 , wherein all of the nucleotides comprise a nucleotide modification.
9 .- 16 . (canceled)
17 . The agent of claim 1 , wherein the nucleotide modification comprises a sugar moiety modification selected from the group consisting of: a 2′-O-methoxyethyl sugar moiety modification, a 2′-methoxy sugar moiety modification, a 2′-O-alkyl sugar moiety modification, and a bicyclic sugar moiety modification.
18 . The agent of claim 17 , wherein the bicyclic sugar moiety modification has a (—CRH—)n group forming a bridge between the 2′ oxygen and the 4′ carbon atoms of the sugar ring, wherein n is 1 or 2 and wherein R is H, CH 3 or CH 3 OCH 3 .
19 .- 20 . (canceled)
21 . The agent of claim 1 , wherein the nucleotide modification comprises an internucleoside linkage modification, wherein the internucleoside linkage modification is a phosphorothioate internucleoside linkage modification.
22 . (canceled)
23 . The agent of claim 1 , wherein the agent is a gapmer comprising a plurality of 2′-deoxynucleotides flanked on each of a 5′ and a 3′ side by a wing segment comprising at least one nucleotide having a sugar moiety modification.
24 . (canceled)
25 . The agent of claim 23 , wherein the sugar moiety modification is selected from the group consisting of a 2′-O-methoxyethyl sugar moiety modification, a 2′-methoxy sugar moiety modification, a 2′-O-alkyl sugar moiety modification, and a bicyclic sugar moiety modification.
26 . The agent of claim 24 , wherein the 5′-wing segment is 1 to 6 nucleotides in length: 2 nucleotides in length: 3 nucleotides in length: 4 nucleotides in length; or 5 nucleotides in length.
27 . The agent of claim 24 , wherein the 3′-wing segment is 1 to 6 nucleotides in length: 2 nucleotides in length: 3 nucleotides in length: 4 nucleotides in length; or 5 nucleotides in length.
28 . The agent of claim 24 , wherein the gap segment is 5 to 14 nucleotides in length; or 10 nucleotides in length.
29 .- 37 . (canceled)
38 . An antisense polynucleotide agent for inhibiting expression of a Programmed cell death 1 ligand 1 (PD-L1) gene, comprising
a gap segment consisting of linked deoxynucleotides;
a 5′-wing segment consisting of linked nucleotides;
a 3′-wing segment consisting of linked nucleotides;
wherein the gap segment is positioned between the 5′-wing segment and the 3′-wing segment and wherein each nucleotide of each wing segment comprises a sugar modification,
wherein the gap segment is ten 2′-deoxynucleotides in length and each of the wing segments is five nucleotides; four nucleotides, three nucleotides, or two nucleotides in length, and
wherein the sugar moiety modification is selected from the group consisting of a 2′-O-methoxyethyl sugar moiety modification, a 2′-methoxy sugar moiety modification, a 2′-O-alkyl sugar moiety modification, and a bicyclic sugar moiety modification.
39 .- 43 . (canceled)
44 . The agent of claim 1 , wherein the agent further comprises a ligand.
45 . The agent of claim 44 , wherein the antisense polynucleotide agent is conjugated to the ligand at the 3′-terminus.
46 . The agent of claim 44 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
47 . The agent of claim 46 , wherein the ligand is
48 . A pharmaceutical composition for inhibiting expression of a Programmed cell death 1 ligand 1 gene comprising the agent of claim 1 .
49 .- 56 . (canceled)
57 . A method of inhibiting expression of a Programmed cell death 1 ligand 1 (PD-L1) gene in a cell, the method comprising:
(a) contacting the cell with the agent of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of a PD-L1_gene, thereby inhibiting expression of the PD-L1_gene in the cell.
58 .- 60 . (canceled)
61 . A method of treating a subject having a disease or disorder that would benefit from reduction in expression of a Programmed cell death 1 ligand 1 (PD-L1) gene, the method comprising administering to the subject a therapeutically effective amount of the agent of claim 1 , thereby treating the subject.
62 .- 75 . (canceled)