NUCLEIC ACID COMPOUNDS FOR INHIBITING MYC GENE EXPRESSION AND USES THEREOF
The present disclosure provides meroduplex ribonucleic acid molecules (mdRNA) capable of decreasing or silencing MYC gene expression. An mdRNA of this disclosure comprises at least three strands that combine to form at least two non-over-lapping double-stranded regions separated by a nick or gap wherein one strand is complementary to a MYC mRNA. In addition, the meroduplex may have at least one uridine substituted with a 5-methyluridine, a nucleoside replaced with a locked nucleic acid, or optionally other modifications, and any combination thereof. Also provided are methods of decreasing expression of a MYC gene in a cell or in a subject to treat a MYC-related disease.
1 - 24 . (canceled)
25 . A meroduplex ribonucleic acid (mdRNA) molecule that down regulates the expression of any one of a human v-myc myelocytomatosis viral oncogene homolog (avian) (MYC) mRNA, comprising a first strand of 15 to 40 nucleotides in length that is complementary to a portion of a human MYC mRNA as set forth in SEQ ID NO:1158, and a second strand and a third strand that is each complementary to non-overlapping regions of the first strand, wherein the second strand and third strand can anneal with the first strand to form at least two double-stranded regions spaced apart by a nick or a gap.
26 . The mdRNA molecule of claim 25 wherein the first strand is 15 to 25 nucleotides in length or 26 to 40 nucleotides in length.
27 . The mdRNA molecule of claim 25 wherein the gap comprises from 1 to 10 unpaired nucleotides.
28 . The mdRNA molecule of claim 25 wherein the double-stranded regions have a combined length of about 15 base pairs to about 40 base pairs.
29 . The mdRNA molecule of claim 25 wherein the mdRNA molecule comprises at least one 5-methyluridine, 2-thioribothymidine, or 2′-O-methyl-5-methyluridine.
30 . The mdRNA molecule of claim 25 wherein the mdRNA molecule comprises at least one locked nucleic acid (LNA) molecule, deoxy nucleotide, G clamp, 2′-sugar modification, modified internucleoside linkage, or any combination thereof.
31 . The mdRNA molecule of claim 25 wherein the mdRNA contains an overhang of one to four nucleotides on at least one 3′-end that is not part of the gap or has a blunt end at one or both ends of the mdRNA.
32 . The mdRNA molecule of claim 25 wherein at least one pyrimidine of the mdRNA molecule is a pyrimidine nucleoside according to Formula I or II:
wherein:
R 1 and R 2 are each independently a —H, —OH, —OCH 3 , —OCH 2 OCH 2 CH 3 , —OCH 2 CH 2 OCH 3 , halogen, substituted or unsubstituted C 1 -C 10 alkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, carboxyalkyl, alkylsulfonylamino, aminoalkyl, dialkylamino, alkylaminoalkyl, dialkylaminoalkyl, haloalkyl, trifluoromethyl, cycloalkyl, (cycloalkyl)alkyl, substituted or unsubstituted C 2 -C 10 alkenyl, substituted or unsubstituted —O-allyl, —O—CH 2 CH═CH 2 , —O—CH═CHCH 3 , substituted or unsubstituted C 2 -C 10 alkynyl, carbamoyl, carbamyl, carboxy, carbonylamino, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, —NH 2 , —NO 2 , —C═, or heterocyclo group,
R 3 and R 4 are each independently a hydroxyl, a protected hydroxyl, a phosphate, or an internucleoside linking group, and
R 5 and R 8 are each independently O or S.
33 . The mdRNA molecule of claim 34 wherein at least one nucleoside is according to Formula I and in which R 1 is methyl and R 2 is —OH or —O-methyl.
34 . The mdRNA molecule of claim 34 wherein at least one R 2 is selected from the group consisting of 2′-O—(C 1 -C 5 ) alkyl, 2′-O-methyl, 2′-OCH 2 OCH 2 CH 3 , 2′-OCH 2 CH 2 OCH 3 , 2′-O-allyl, and fluoro.
35 . The mdRNA molecule of claim 25 wherein the first strand is 19 to 23 nucleotides in length and is complementary to a human MYC nucleic acid sequence as set forth in any one of SEQ ID NOS:1159-1288.
36 . The mdRNA molecule of claim 25 wherein the first strand is 25 to 29 nucleotides in length and is complementary to a human MYC nucleic acid sequence as set forth in any one of SEQ ID NOS:1289-1385.
37 . A method for reducing the expression of a human MYC gene, comprising administering an mdRNA molecule of claim 25 to a cell expressing a human MYC gene, wherein the mdRNA molecule reduces the expression of the human MYC gene in the cell.
38 . The method according to claim 37 wherein the cell is a human cell.
39 . A double-stranded ribonucleic acid (dsRNA) molecule that down regulates the expression of any one of a human v-myc myelocytomatosis viral oncogene homolog (avian) (MYC) mRNA, comprising a first strand of 15 to 40 nucleotides in length that is complementary to a portion of a human MYC mRNA as set forth in SEQ ID NO:1158 and a second strand that is complementary to the first strand.
40 . The dsRNA molecule of claim 39 wherein the first strand is from 15 to 25 nucleotides in length or 26 to 40 nucleotides in length.
41 . The dsRNA molecule of claim 39 wherein the dsRNA molecule has a blunt end at one or both ends of the dsRNA.
42 . The dsRNA molecule of claim 39 wherein the dsRNA molecule has a 3′-end overhang of one to four nucleotides at one or both ends of the dsRNA.
43 . The dsRNA molecule of claim 39 wherein the dsRNA molecule comprises at least one 5-methyluridine, 2-thioribothymidine, or 2′-O-methyl-5-methyluridine.
44 . The dsRNA molecule of claim 39 wherein the dsRNA molecule comprises at least one locked nucleic acid (LNA) molecule, deoxy nucleotide, G clamp, 2′-sugar modification, modified internucleoside linkage, or any combination thereof.
45 . The dsRNA molecule of claim 39 wherein the dsRNA molecule has a 5′-terminal end comprising a hydroxyl or a phosphate.
46 . The dsRNA molecule of claim 39 wherein at least one pyrimidine of the dsRNA molecule comprises a pyrimidine nucleoside according to Formula I or II:
wherein:
R 1 and R 2 are each independently a —H, —OH, —OCH 3 , —OCH 2 OCH 2 CH 3 , —OCH 2 CH 2 OCH 3 , halogen, substituted or unsubstituted C 1 -C 10 alkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, carboxyalkyl, alkylsulfonylamino, aminoalkyl, dialkylamino, alkylaminoalkyl, dialkylaminoalkyl, haloalkyl, trifluoromethyl, cycloalkyl, (cycloalkyl)alkyl, substituted or unsubstituted C 2 -C 10 alkenyl, substituted or unsubstituted —O-allyl, —O—CH 2 CH═CH 2 , —O—CH═CHCH 3 , substituted or unsubstituted C 2 -C 10 alkynyl, carbamoyl, carbamyl, carboxy, carbonylamino, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, —NH 2 , —NO 2 , —C≡N, or heterocyclo group,
R 3 and R 4 are each independently a hydroxyl, a protected hydroxyl, a phosphate, or an internucleoside linking group, and
R 5 and R 8 are each independently O or S.
47 . The dsRNA molecule of claim 46 wherein at least one nucleoside is according to Formula I and in which R 1 is methyl and R 2 is —OH or —O-methyl.
48 . The dsRNA molecule of claim 46 wherein at least one R 2 is selected from the group consisting of 2′-O—(C 1 -C 5 ) alkyl, 2′-O-methyl, 2′-OCH 2 OCH 2 CH 3 , 2′-OCH 2 CH 2 OCH 3 , 2′-O-allyl, and 2′-fluoro.
49 . A method for reducing the expression of a human MYC gene, comprising administering a dsRNA molecule of claim 39 to a cell expressing a human MYC gene, wherein the dsRNA molecule reduces the expression of the human MYC gene in the cell.
50 . The method according to claim 49 wherein the cell is a human cell.
51 . The dsRNA molecule of claim 39 wherein the first strand is 19 to 23 nucleotides in length and is complementary to a human MYC nucleic acid sequence as set forth in any one of SEQ ID NOS:1159-1288.
52 . The dsRNA molecule of claim 39 wherein the first strand is 25 to 29 nucleotides in length and is complementary to a human MYC nucleic acid sequence as set forth in any one of SEQ ID NOS:1289-1385.