XANTHINE DEHYDROGENASE (XDH) IRNA COMPOSITIONS AND METHODS OF USE THEREOF
The present invention relates to RNAi agents, e.g., double stranded RNAi agents, targeting a xanthine dehydrogenase (XDH) gene, and methods of using such double stranded RNAi agents to inhibit expression of an XDH gene and methods of treating subjects having an XDH-associated disease.
1 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a xanthine dehydrogenase (XDH) gene, wherein said dsRNA agent comprises a sense strand and an antisense strand, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:1 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:2.
2 . The dsRNA of claim 1 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of any one of nucleotides 490-512, 86-104, 273-291, 339-358, 410-428, 444-462, 490-509, 493-512, 888-907, 936-954, 969-987, 1105-1123, 1158-1176, 1242-1260, 1326-1344, 1357-1412, 1357-1379, 1357-1376, 1360-1379, 1378-1412, 1378-1396, 1394-1412, 1481-1499, 1515-1548, 1515-1533, 1530-1548, 1718-1736, 1783-1802, 1854-1890, 1854-1872, 1872-1890, 2053-2072, 2077-2096, 2137-2160, 2137-2156, 2142-2160, 2173-2206, 2173-2192, 2177-2195, 2184-2203, 2187-2206, 2314-2332, 2567-2604, 2567-2585, 2585-2604, 2620-2640, 2620-2639, 2621-2640, 2722-2740, 2891-2909, 2941-2975, 2941-2959, 2956-2975, 2993-3011, 3025-3061, 3025-3044, 3042-3061, 3062-3110, 3062-3080, 3079-3097, 3091-3110, 3112-3147, 3112-3130, 3129-3147, 3197-3215, 3247-3265, 3316-3334, 3366-3384, 3487-3520, 3487-3505, 3502-3520, 3606-3624, 3672-3690, 3891-3930, 3891-3910, 3893-3912, 3912-3930, 4063-4081, 4114-4132, 4152-4171, 4200-4218, 4300-4337, 4300-4319, 4303-4321, 4319-4337, 4386-4404, 4519-4538, 4541-4559, 4618-4637, or 4703-4722 of the nucleotide sequence of SEQ ID NO:1.
3 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a xanthine dehydrogenase (XDH) gene, wherein said dsRNA agent comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity which comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences listed in any one of Tables 3, 4, 6, and 7.
4 . The dsRNA agent of claim 3 , wherein the region of complementarity of the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences of any one of duplexes AD-70016, AD-70033, AD-70053, AD-70050, AD-70051, AD-72007, AD-71990, AD-70049, AD-71993, AD-70042, AD-70052, AD-70018, AD-70055, AD-71831, AD-71810, AD-70023, AD-70035, AD-70044, AD-71820, AD-70027, AD-70034, AD-71837, AD-71861, AD-71998, AD-70020, AD-70026, AD-70030, AD-71801, AD-72003, AD-70025, AD-70021, AD-70029, AD-71834, AD-71765, AD-70043, AD-71840, AD-72006, AD-71844, AD-71779, AD-71830, AD-71952, AD-71766, AD-71950, AD-70022, AD-71833, AD-71823, AD-71847, AD-71900, AD-72014, AD-72015, AD-70037, AD-71982, AD-71787, AD-71894, AD-70048, AD-71887, AD-70028, AD-71980, AD-71826, AD-71855, AD-71778, AD-71757, AD-72012, AD-71854, AD-71890, AD-70038, AD-71865, AD-71933, AD-71942, AD-71901, AD-71878, AD-71905, or AD-71914.
5 . The dsRNA agent of any one of claims 1 - 4 , wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of of any of the nucleotide sequences listed in any one of Tables 3, 4, 6, and 7.
6 . The dsRNA agent of any one of claims 1 - 4 , wherein said dsRNA agent comprises at least one nucleotide comprising a nucleotide modification.
7 . The dsRNA agent of any one of claims 1 - 4 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification.
8 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a xanthine dehydrogenase (XDH) gene, wherein said dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:1 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:2,
wherein substantially all of the nucleotides of said sense strand and substantially 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.
9 . The dsRNA agent of claim 8 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of any one of nucleotides 490-512, 86-104, 273-291, 339-358, 410-428, 444-462, 490-509, 493-512, 888-907, 936-954, 969-987, 1105-1123, 1158-1176, 1242-1260, 1326-1344, 1357-1412, 1357-1379, 1357-1376, 1360-1379, 1378-1412, 1378-1396, 1394-1412, 1481-1499, 1515-1548, 1515-1533, 1530-1548, 1718-1736, 1783-1802, 1854-1890, 1854-1872, 1872-1890, 2053-2072, 2077-2096, 2137-2160, 2137-2156, 2142-2160, 2173-2206, 2173-2192, 2177-2195, 2184-2203, 2187-2206, 2314-2332, 2567-2604, 2567-2585, 2585-2604, 2620-2640, 2620-2639, 2621-2640, 2722-2740, 2891-2909, 2941-2975, 2941-2959, 2956-2975, 2993-3011, 3025-3061, 3025-3044, 3042-3061, 3062-3110, 3062-3080, 3079-3097, 3091-3110, 3112-3147, 3112-3130, 3129-3147, 3197-3215, 3247-3265, 3316-3334, 3366-3384, 3487-3520, 3487-3505, 3502-3520, 3606-3624, 3672-3690, 3891-3930, 3891-3910, 3893-3912, 3912-3930, 4063-4081, 4114-4132, 4152-4171, 4200-4218, 4300-4337, 4300-4319, 4303-4321, 4319-4337, 4386-4404, 4519-4538, 4541-4559, 4618-4637, or 4703-4722 of the nucleotide sequence of SEQ ID NO:1.
10 . The dsRNA agent of claim 8 or 9 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification.
11 . The dsRNA agent of any one of claims 6 - 10 , wherein the modified nucleotides are independently selected from the group consisting of a deoxy-nucleotide, a 3′-terminal deoxy-thymine (dT) nucleotide, a 2′-O-methyl modified nucleotide, a 2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an unlocked nucleotide, a conformationally restricted nucleotide, a constrained ethyl nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2′-O-allyl-modified nucleotide, 2′-C-alkyl-modified nucleotide, 2′-hydroxly-modified nucleotide, a 2′-methoxyethyl modified nucleotide, a 2′-O-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, a non-natural base comprising nucleotide, a tetrahydropyran modified nucleotide, a 1,5-anhydrohexitol modified nucleotide, a cyclohexenyl modified nucleotide, a nucleotide comprising a phosphorothioate group, a nucleotide comprising a methylphosphonate group, a nucleotide comprising a 5′-phosphate, and a nucleotide comprising a 5′-phosphate mimic.
12 . The dsRNA agent of claim 11 , wherein said modified nucleotides comprise a short sequence of 3′-terminal deoxy-thymine nucleotides (dT).
13 . The dsRNA agent of claim 3 or 4 , wherein the region of complementarity is at least 17 nucleotides in length.
14 . The dsRNA agent of claim 3 or 4 , wherein the region of complementarity is 19 to 21 nucleotides in length.
15 . The dsRNA agent of claim 14 , wherein the region of complementarity is 19 nucleotides in length.
16 . The dsRNA agent of any one of claims 1 - 4 and 8 , wherein each strand is no more than 30 nucleotides in length.
17 . The dsRNA agent of any one of claims 1 - 4 and 8 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.
18 . The dsRNA agent of any one of claims 1 - 4 and 8 , wherein at least one strand comprises a 3′ overhang of at least 2 nucleotides.
19 . The dsRNA agent of any one of claims 1 - 4 further comprising a ligand.
20 . The dsRNA agent of claim 19 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA agent.
21 . The dsRNA agent of claim 8 or 19 , wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.
22 . The dsRNA agent of claim 21 , wherein the ligand is
23 . The dsRNA agent of claim 22 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
and, wherein X is O or S.
24 . The dsRNA agent of claim 23 , wherein the X is O.
25 . The dsRNA agent of claim 3 or 4 , wherein the region of complementarity comprises any one of the antisense nucleotide sequences of any one of Tables 3, 4, 6 and 7.
26 . The dsRNA agent of claim 3 or 4 , wherein the region of complementarity consists of any one of the antisense nucleotide sequences of any one of Tables 3, 4, 6 and 7.
27 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of a xanthine dehydrogenase (XDH) gene, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
antisense: 3′n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) i -N a ′-n q ′5′ (III)
wherein:
i, j, k, and 1 are each independently 0 or 1;
p, p′, q, and q′ are each independently 0-6;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof;
each n p , n p ′, n q , and n q ′, each of which may or may not be present, independently represents an overhang nucleotide;
XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides;
modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′; and
wherein the sense strand is conjugated to at least one ligand.
28 . The dsRNA agent of claim 27 , wherein i is 0; j is 0; i is 1; j is 1; both i and j are 0; or both i and j are 1.
29 . The dsRNA agent of claim 27 , wherein k is 0; l is 0; k is 1; l is 1; both k and l are 0; or both k and l are 1.
30 . The dsRNA agent of claim 27 , wherein XXX is complementary to X′X′X′, YYY is complementary to Y′Y′Y′, and ZZZ is complementary to Z′Z′Z′.
31 . The dsRNA agent of claim 27 , wherein the YYY motif occurs at or near the cleavage site of the sense strand.
32 . The dsRNA agent of claim 27 , wherein the Y′Y′Y′ motif occurs at the 11, 12 and 13 positions of the antisense strand from the 5′-end.
33 . The dsRNA agent of claim 32 , wherein the Y′ is 2′-O-methyl.
34 . The dsRNA agent of claim 27 , wherein formula (III) is represented by formula (IIIa):
sense: 5′n p -N a -YYY-N a -n q 3′
antisense: 3′n p′ -N a′ -Y′Y′Y′-N a′ -n q′ 5′ (IIIa).
35 . The dsRNA agent of claim 27 , wherein formula (III) is represented by formula (IIIb):
sense: 5′n p -N a -YYY-N b -ZZZ-N a -n q 3′
antisense: 3′n p′ -N a′ -Y′Y′Y′-N b′ -Z′Z′Z′-N a′ -n q′ 5′ (IIIb)
wherein each N b and N b ′ independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides.
36 . The dsRNA agent of claim 27 , wherein formula (III) is represented by formula (IIIc):
sense: 5′n p -N a -XXX-N b -YYY-n q 3′
antisense: 3′n p′ -N a′ -X′X′X′-N b′ -Y′Y′Y′-N a′ -n q′ 5′ (IIIc)
wherein each N b and N b ′ independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides.
37 . The dsRNA agent of claim 27 , wherein formula (III) is represented by formula (IIId):
sense: 5′n p -N a -XXX-N b -YYY-N b -ZZZ-N a -n q 3′
antisense: 3′n p′ -N a′ -X′X′X′-N b′ -Y′Y′Y′-N b′ -Z′Z′Z′-N a′ -n q′ 5′ (IIId)
wherein each N b and N b ′ independently represents an oligonucleotide sequence comprising 1-5 modified nucleotides and each N a and N a ′ independently represents an oligonucleotide sequence comprising 2-10 modified nucleotides.
38 . The dsRNA agent of claim 8 or 27 , wherein the double stranded region is 15-30 nucleotide pairs in length.
39 . The dsRNA agent of claim 38 , wherein the double stranded region is 17-23 nucleotide pairs in length.
40 . The dsRNA agent of claim 38 , wherein the double stranded region is 17-25 nucleotide pairs in length.
41 . The dsRNA agent of claim 38 , wherein the double stranded region is 23-27 nucleotide pairs in length.
42 . The dsRNA agent of claim 38 , wherein the double stranded region is 19-21 nucleotide pairs in length.
43 . The dsRNA agent of claim 8 or 27 , wherein the double stranded region is 21-23 nucleotide pairs in length.
44 . The dsRNA agent of claim 27 , wherein each strand has 15-30 nucleotides.
45 . The dsRNA agent of claim 8 or 27 , wherein each strand has 19-30 nucleotides.
46 . The dsRNA agent of claim 8 or 27 , wherein the modifications on the nucleotides are selected from the group consisting of LNA, HNA, CeNA, 2′-methoxyethyl, 2′-O-alkyl, 2′-O-allyl, 2′-C-allyl, 2′-fluoro, 2′-deoxy, 2′-hydroxyl, and combinations thereof.
47 . The dsRNA agent of claim 46 , wherein the modifications on the nucleotides are 2′-O-methyl or 2′-fluoro modifications.
48 . The dsRNA agent of claim 8 or 27 , wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
49 . The dsRNA agent of claim 27 , the ligand is
50 . The dsRNA agent of claim 27 , wherein the ligand is attached to the 3′ end of the sense strand.
51 . The dsRNA agent of claim 50 , wherein the RNAi agent is conjugated to the ligand as shown in the following schematic
52 . The dsRNA agent of claim 8 or 27 , wherein said agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.
53 . The dsRNA agent of claim 52 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 3′-terminus of one strand.
54 . The dsRNA agent of claim 53 , wherein said strand is the antisense strand.
55 . The dsRNA agent of claim 53 , wherein said strand is the sense strand.
56 . The dsRNA agent of claim 52 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the 5′-terminus of one strand.
57 . The dsRNA agent of claim 56 , wherein said strand is the antisense strand.
58 . The dsRNA agent of claim 56 , wherein said strand is the sense strand.
59 . The dsRNA agent of claim 52 , wherein the phosphorothioate or methylphosphonate internucleotide linkage is at the both the 5′- and 3′-terminus of one strand.
60 . The dsRNA agent of claim 59 , wherein said strand is the antisense strand.
61 . The dsRNA agent of claim 8 or 27 , wherein the base pair at the 1 position of the 5′-end of the antisense strand of the duplex is an AU base pair.
62 . The dsRNA agent of claim 27 , wherein the Y nucleotides contain a 2′-fluoro modification.
63 . The dsRNA agent of claim 27 , wherein the Y′ nucleotides contain a 2′-O-methyl modification.
64 . The dsRNA agent of claim 27 , wherein p′>0.
65 . The dsRNA agent of claim 27 , wherein p′=2.
66 . The dsRNA agent of claim 65 , wherein q′=0, p=0, q=0, and p′ overhang nucleotides are complementary to the target mRNA.
67 . The dsRNA agent of claim 65 , wherein q′=0, p=0, q=0, and p′ overhang nucleotides are non-complementary to the target mRNA.
68 . The dsRNA agent of claim 59 , wherein the sense strand has a total of 21 nucleotides and the antisense strand has a total of 23 nucleotides.
69 . The dsRNA agent of any one of claims 64 - 68 , wherein at least one n p ′ is linked to a neighboring nucleotide via a phosphorothioate linkage.
70 . The dsRNA agent of claim 69 , wherein all n p ′ are linked to neighboring nucleotides via phosphorothioate linkages.
71 . The dsRNA agent of claim 27 , wherein said dsRNA agent is selected from the group of dsRNA agents listed in any one of Tables 3, 4, 6, and 7.
72 . The dsRNA agent of claim 24 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification.
73 . A double stranded ribonucleic acid (dsRNA) agent capable for inhibiting the expression of a xanthine dehydrogenase (XDH) gene in a cell, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
antisense: 3′n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) i -N a ′-n q ′5′ (III)
wherein:
i, j, k, and 1 are each independently 0 or 1;
p, p′, q, and q′ are each independently 0-6;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof;
each n p , n p ′, n q , and n q ′, each of which may or may not be present independently represents an overhang nucleotide;
XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2′-O-methyl or 2′-fluoro modifications;
modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′; and
wherein the sense strand is conjugated to at least one ligand.
74 . A double stranded ribonucleic acid (dsRNA) agent capable for inhibiting the expression of a xanthine dehydrogenase (XDH) gene in a cell, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
antisense: 3′n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) i -N a ′-n q ′5′ (III)
wherein:
i, j, k, and 1 are each independently 0 or 1;
each n p , n q , and n q ′, each of which may or may not be present, independently represents an overhang nucleotide;
p, q, and q′ are each independently 0-6;
n p ′>0 and at least one n p ′ is linked to a neighboring nucleotide via a phosphorothioate linkage;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof;
XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2′-O-methyl or 2′-fluoro modifications;
modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′; and
wherein the sense strand is conjugated to at least one ligand.
75 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of a xanthine dehydrogenase (XDH) gene in a cell, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
antisense: 3′n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) i -N a ′-n q ′5′ (III)
wherein:
i, j, k, and 1 are each independently 0 or 1;
each n p , n q , and n q ′, each of which may or may not be present, independently represents an overhang nucleotide;
p, q, and q′ are each independently 0-6;
n p ′>0 and at least one n p ′ is linked to a neighboring nucleotide via a phosphorothioate linkage;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof;
XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2′-O-methyl or 2′-fluoro modifications;
modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′; and
wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
76 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of a xanthine dehydrogenase (XDH) gene in a cell, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -(XXX) i -N b -YYY-N b -(ZZZ) j -N a -n q 3′
antisense: 3′n p ′-N a ′-(X′X′X′) k -N b ′-Y′Y′Y′-N b ′-(Z′Z′Z′) i -N a ′-n q ′5′ (III)
wherein:
i, j, k, and 1 are each independently 0 or 1;
each n p , n q , and n q ′, each of which may or may not be present, independently represents an overhang nucleotide;
p, q, and q′ are each independently 0-6;
n p ′>0 and at least one n p ′ is linked to a neighboring nucleotide via a phosphorothioate linkage;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
each N b and N b ′ independently represents an oligonucleotide sequence comprising 0-10 nucleotides which are either modified or unmodified or combinations thereof;
XXX, YYY, ZZZ, X′X′X′, Y′Y′Y′, and Z′Z′Z′ each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2′-O-methyl or 2′-fluoro modifications;
modifications on N b differ from the modification on Y and modifications on N b ′ differ from the modification on Y′;
wherein the sense strand comprises at least one phosphorothioate linkage; and
wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
77 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting the expression of a xanthine dehydrogenase (XDH) gene in a cell, wherein said dsRNA agent comprises a sense strand complementary to an antisense strand, wherein said antisense strand comprises a region complementary to part of an mRNA encoding XDH, wherein each strand is about 14 to about 30 nucleotides in length, wherein said dsRNA agent is represented by formula (III):
sense: 5′n p -N a -YYY-N a -n q 3′
antisense: 3′n p ′-N a ′-Y′Y′Y′-N a ′-n q ′5′ (IIIa)
wherein:
each n p , n q , and n q ′, each of which may or may not be present, independently represents an overhang nucleotide;
p, q, and q′ are each independently 0-6;
n p ′>0 and at least one n p ′ is linked to a neighboring nucleotide via a phosphorothioate linkage;
each N a and N a ′ independently represents an oligonucleotide sequence comprising 0-25 nucleotides which are either modified or unmodified or combinations thereof, each sequence comprising at least two differently modified nucleotides;
YYY and Y′Y′Y′ each independently represent one motif of three identical modifications on three consecutive nucleotides, and wherein the modifications are 2′-O-methyl or 2′-fluoro modifications;
wherein the sense strand comprises at least one phosphorothioate linkage; and
wherein the sense strand is conjugated to at least one ligand, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.
78 . The dsRNA agent of any one of claims 73 - 77 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of nucleotides 490-512, 86-104, 273-291, 339-358, 410-428, 444-462, 490-509, 493-512, 888-907, 936-954, 969-987, 1105-1123, 1158-1176, 1242-1260, 1326-1344, 1357-1412, 1357-1379, 1357-1376, 1360-1379, 1378-1412, 1378-1396, 1394-1412, 1481-1499, 1515-1548, 1515-1533, 1530-1548, 1718-1736, 1783-1802, 1854-1890, 1854-1872, 1872-1890, 2053-2072, 2077-2096, 2137-2160, 2137-2156, 2142-2160, 2173-2206, 2173-2192, 2177-2195, 2184-2203, 2187-2206, 2314-2332, 2567-2604, 2567-2585, 2585-2604, 2620-2640, 2620-2639, 2621-2640, 2722-2740, 2891-2909, 2941-2975, 2941-2959, 2956-2975, 2993-3011, 3025-3061, 3025-3044, 3042-3061, 3062-3110, 3062-3080, 3079-3097, 3091-3110, 3112-3147, 3112-3130, 3129-3147, 3197-3215, 3247-3265, 3316-3334, 3366-3384, 3487-3520, 3487-3505, 3502-3520, 3606-3624, 3672-3690, 3891-3930, 3891-3910, 3893-3912, 3912-3930, 4063-4081, 4114-4132, 4152-4171, 4200-4218, 4300-4337, 4300-4319, 4303-4321, 4319-4337, 4386-4404, 4519-4538, 4541-4559, 4618-4637, or 4703-4722 of the nucleotide sequence of SEQ ID NO:1.
79 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a xanthine dehydrogenase (XDH) gene in a cell,
wherein said dsRNA agent comprises a sense strand and an antisense strand forming a double stranded region,
wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:1 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO:2,
wherein substantially all of the nucleotides of said sense strand comprise a modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said sense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus,
wherein substantially all of the nucleotides of said antisense strand comprise a nucleotide modification selected from the group consisting of a 2′-O-methyl modification and a 2′-fluoro modification,
wherein said antisense strand comprises two phosphorothioate internucleotide linkages at the 5′-terminus and two phosphorothioate internucleotide linkages at the 3′-terminus, and
wherein said sense strand is conjugated to one or more GalNAc derivatives attached through a branched bivalent or trivalent linker at the 3′-terminus.
80 . The dsRNA agent of claim 79 , wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of any one of nucleotides 490-512, 86-104, 273-291, 339-358, 410-428, 444-462, 490-509, 493-512, 888-907, 936-954, 969-987, 1105-1123, 1158-1176, 1242-1260, 1326-1344, 1357-1412, 1357-1379, 1357-1376, 1360-1379, 1378-1412, 1378-1396, 1394-1412, 1481-1499, 1515-1548, 1515-1533, 1530-1548, 1718-1736, 1783-1802, 1854-1890, 1854-1872, 1872-1890, 2053-2072, 2077-2096, 2137-2160, 2137-2156, 2142-2160, 2173-2206, 2173-2192, 2177-2195, 2184-2203, 2187-2206, 2314-2332, 2567-2604, 2567-2585, 2585-2604, 2620-2640, 2620-2639, 2621-2640, 2722-2740, 2891-2909, 2941-2975, 2941-2959, 2956-2975, 2993-3011, 3025-3061, 3025-3044, 3042-3061, 3062-3110, 3062-3080, 3079-3097, 3091-3110, 3112-3147, 3112-3130, 3129-3147, 3197-3215, 3247-3265, 3316-3334, 3366-3384, 3487-3520, 3487-3505, 3502-3520, 3606-3624, 3672-3690, 3891-3930, 3891-3910, 3893-3912, 3912-3930, 4063-4081, 4114-4132, 4152-4171, 4200-4218, 4300-4337, 4300-4319, 4303-4321, 4319-4337, 4386-4404, 4519-4538, 4541-4559, 4618-4637, or 4703-4722 of the nucleotide sequence of SEQ ID NO:1.
81 . The dsRNA agent of claim 79 , wherein all of the nucleotides of said sense strand and all of the nucleotides of said antisense strand comprise a nucleotide modification.
82 . The dsRNA agent of claim 79 , wherein each strand has 19-30 nucleotides.
83 . A cell containing the dsRNA agent of any one of claims 1 - 4 , 8 , 27 , 73 - 77 , and 79 .
84 . A vector encoding at least one strand of a dsRNA agent, wherein said dsRNA agent comprises a region of complementarity to at least a part of an mRNA encoding a xanthine dehydrogenase (XDH) gene, wherein said dsRNA agent is 30 base pairs or less in length, and wherein said dsRNA agent targets said mRNA for cleavage.
85 . A cell comprising the vector of claim 84 .
86 . A pharmaceutical composition for inhibiting expression of an XDH gene comprising the dsRNA agent of any one of claims 1 - 4 , 8 , 27 , 73 - 77 , and 79 or the vector of claim 84 .
87 . The pharmaceutical composition of claim 86 , wherein the dsRNA agent is administered in an unbuffered solution.
88 . The pharmaceutical composition of claim 87 , wherein said unbuffered solution is saline or water.
89 . The pharmaceutical composition of claim 86 , wherein said dsRNA agent is administered with a buffer solution.
90 . The pharmaceutical composition of claim 89 , wherein said buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
91 . The pharmaceutical composition of claim 89 , wherein said buffer solution is phosphate buffered saline (PBS).
92 . A pharmaceutical composition comprising the dsRNA agent of any one of claims 1 - 2 , and a lipid formulation.
93 . A method of inhibiting xanthine dehydrogenase (XDH) expression in a cell, the method comprising contacting the cell with the dsRNA agent of any one of claims claims 1 - 4 , 8 , 27 , 73 - 77 , and 79 or a pharmaceutical composition of any one of claims 86 - 92 , thereby inhibiting expression of the XDH gene in the cell.
94 . The method of claim 93 , wherein said cell is within a subject.
95 . The method of claim 94 , wherein the subject is a human.
96 . The method of any one of claims 93 - 95 , wherein the XDH expression is inhibited by at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 98%, or to below an assay detection level.
97 . A method of treating a subject having a disease or disorder that would benefit from reduction in XDH expression, the method comprising administering to the subject a therapeutically effective amount of the dsRNA agent of any one of claims 1 - 4 , 8 , 27 , 73 - 77 , and 79 or a pharmaceutical composition of any one of claims 86 - 92 , thereby treating said subject.
98 . A method of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in XDH expression, the method comprising administering to the subject a prophylactically effective amount of the dsRNA agent of any one of claims 1 - 4 , 8 , 27 , 73 - 77 , and 79 or a pharmaceutical composition of any one of claims 86 - 92 , thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in XDH expression.
99 . The method of claim 97 or 98 , wherein the administration of the double stranded RNAi agent to the subject decreases serum uric acid.
100 . The method of claim 97 or 98 , wherein the disorder is an XDH-associated disease.
101 . The method of claim 100 , wherein the XDH-associated disease is hyperuricemia.
102 . The method of claim 100 , wherein the XDH-associated disease is gout.
103 . The method of claim 100 , wherein the XDH-associated disease is selected from the group consisting of NAFLD, NASH, metabolic disorder, insulin resistance, cardiovascular disease, type 2 diabetes, and a condition linked to oxidative stress.
104 . The method of any of claims 97 - 103 , wherein the subject has renal insufficiency.
105 . The method of any one of claims 97 - 104 , further comprising administering an additional agent to the subject for the treatment of an XDH-associated disease.
106 . The method of any one of claims 97 - 105 , wherein the dsRNA agent is administered to the subject at a dose of about 0.01 mg/kg to about 50 mg/kg.
107 . The method of any one of claims 97 - 106 , wherein the dsRNA agent is administered to the subject subcutaneously.
108 . The method of claim 97 or 98 , further comprising measuring uric acid level in said subject.
109 . The method of any one of claims 97 - 108 , wherein the subject is human.