Methods and compositions for the specific inhibition of myc by double-stranded rna

Abstract

This invention relates to compounds, compositions, and methods useful for reducing MYC target RNA and protein levels via use of dsRNAs, e.g., Dicer substrate siRNA (DsiRNA) agents.

We claim: 
     
         1 . A double stranded nucleic acid (dsNA) comprising first and second nucleic acid strands, wherein said first strand and said second strand of said dsNA form a duplex region of 15-35 base pairs and wherein said second oligonucleotide strand comprises the complement of SEQ ID NO: 1587 and wherein said dsNA reduces MYC target mRNA expression when said double stranded nucleic acid is introduced into a mammalian cell. 
     
     
         2 . The dsNA of  claim 1 , wherein said second strand of said dsNA comprises SEQ ID NO: 606. 
     
     
         3 . The dsNA of  claim 1 , wherein said dsNA comprises a modified nucleotide. 
     
     
         4 . The dsNA of  claim 1 , wherein said modified nucleotide residue is selected from the group consisting of 2′-O-methyl, 2′-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH2-O-2′-bridge, 4′-(CH2)2-O-2′-bridge, 2′-LNA, 2′-amino and 2′-O—(N-methlycarbamate). 
     
     
         5 . The dsNA of  claim 1 , wherein said dsNA comprises a tetraloop. 
     
     
         6 . The dsNA of  claim 1  comprising a modified nucleotide selected from the group consisting of a dideoxyribonucleotide, an acyclonucleotide, a 3′-deoxyadenosine (cordycepin), a 3′-azido-3′-deoxythymidine (AZT), a 2′,3′-dideoxyinosine (ddI), a 2′,3′-dideoxy-3′-thiacytidine (3TC), a 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T), a monophosphate nucleotide of 3′-azido-3′-deoxythymidine (AZT), a 2′,3′-dideoxy-3′-thiacytidine (3TC) and a monophosphate nucleotide of 2′,3′-didehydro-2′,3′-dideoxythymidine (d4T), a 4-thiouracil, a 5-bromouracil, a 5-iodouracil, a 5-(3-aminoallyl)-uracil, a 2′-O-alkyl ribonucleotide, a 2′-O-methyl ribonucleotide, a 2′-amino ribonucleotide, a 2′-fluoro ribonucleotide, and a locked nucleic acid. 
     
     
         7 . The dsNA of  claim 1  comprising a phosphate backbone modification selected from the group consisting of a phosphonate, a phosphorothioate and a phosphotriester. 
     
     
         8 . The dsNA of  claim 1  comprising a modification selected from the group consisting of a morpholino nucleic acid and a peptide nucleic acid (PNA). 
     
     
         9 . The dsNA of  claim 1  comprising a duplex region of 19-35 base pairs. 
     
     
         10 . The dsNA of  claim 1  comprising a duplex region of 19-25 base pairs. 
     
     
         11 . The dsNA of  claim 1 , wherein said first strand is 25-50 nucleotides in length. 
     
     
         12 . The dsNA of  claim 1 , wherein said second strand is 25-50 nucleotides in length. 
     
     
         13 . The dsNA of  claim 1 , wherein said first strand comprises SEQ ID NO: 2568. 
     
     
         14 . A nucleic acid molecule comprising a sense region and an antisense region, wherein the nucleic acid molecule comprises RNA, the sense region and the antisense region together form a duplex region of 25-35 base pairs and the antisense region comprises the complement of SEQ ID NO: 1587. 
     
     
         15 . The nucleic acid molecule of  claim 14 , wherein said antisense region comprises SEQ ID NO: 606. 
     
     
         16 . The nucleic acid molecule of  claim 14 , wherein said sense region comprises SEQ ID NO: 2568. 
     
     
         17 . The nucleic acid molecule of  claim 14 , wherein the nucleic acid molecule is a double stranded nucleic acid (dsNA) comprising first and second nucleic acid strands. 
     
     
         18 . The dsNA of  claim 14 , wherein said first strand is 25-50 nucleotides in length. 
     
     
         19 . The dsNA of  claim 14 , wherein said second strand is 25-50 nucleotides in length. 
     
     
         20 . The dsNA of  claim 14 , wherein said first strand comprises SEQ ID NO: 2568. 
     
     
         21 . A dsNA comprising first and second nucleic acid strands, wherein said first strand and said second strand of said dsNA form a duplex region of at least 25 base pairs and wherein said second oligonucleotide strand comprises the complement of SEQ ID NO: 1587 and wherein said dsNA reduces MYC target mRNA expression when said double stranded nucleic acid is introduced into a mammalian cell. 
     
     
         22 . A method for reducing expression of a target MYC gene in a mammalian cell comprising contacting a mammalian cell in vitro with the dsNA of  claim 1  in an amount sufficient to reduce expression of a target MYC mRNA in said cell. 
     
     
         23 . A method for reducing expression of a target MYC mRNA in a mammal comprising administering the dsNA of  claim 1  to a mammal in an amount sufficient to reduce expression of a target MYC mRNA in the mammal. 
     
     
         24 . A method for reducing tumor burden in a mammal comprising administering the dsNA of  claim 1  to a mammal in an amount sufficient to reduce tumor burden in said mammal. 
     
     
         25 . The method of  claim 1 , wherein said tumor is a hepatocellular carcinoma. 
     
     
         26 . The method of  claim 1 , wherein said tumor burden is reduced by 50-80%, as compared to a suitable control. 
     
     
         27 . The method of  claim 1 , wherein said dsNA is formulated in a lipid nanoparticle (LNP). 
     
     
         28 . A formulation comprising the dsNA of  claim 1 , wherein said dsNA is present in an amount effective to reduce target MYC mRNA levels when said dsNA is introduced into a cell of a mammalian subject by an amount (expressed by %) selected from the group consisting of at least 10%, at least 50% and at least 80-90%. 
     
     
         29 . The formulation of  claim 1 , wherein said effective amount is a dosage selected from the group consisting of 1 microgram to 5 milligrams per kilogram of said subject per day, 100 micrograms to 0.5 milligrams per kilogram, 0.001 to 0.25 milligrams per kilogram, 0.01 to 20 micrograms per kilogram, 0.01 to 10 micrograms per kilogram, 0.10 to 5 micrograms per kilogram, and 0.1 to 2.5 micrograms per kilogram. 
     
     
         30 . An isolated mammalian cell containing the dsNA of  claim 1 . 
     
     
         31 . A pharmaceutical composition comprising the dsNA of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         32 . A kit comprising the dsNA of  claim 1  and instructions for its use.