PDGF and VEGF aptamers having improved stability and their use in treating PDGF and VEGF mediated diseases and disorders
Aptamers having improved stability against nucleases that bind PDGF and aptamers that bind VEGF are provided. In addition, aptamer constructs comprising a PDGF aptamer and a VEGF aptamer are provided. Pharmaceutical compositions comprising the aptamers and aptamer constructs are provided, as well as methods of treating conditions using the aptamers and aptamer constructs.
1. A nucleic acid molecule comprising the nucleic sequence
5′-C-C-G-Z-Z-C-A-A-G-Z-G-C-Z-Z-G-Z-A-G-G-A-Z-Z-Z-A-A-A-Z-G-G-3′ (SEQ ID NO: 831),
wherein Z is a C-5 modified pyrimidine, and at least one of positions 24, 25 and 26 of the nucleic acid molecule comprise a phosphorothioate linkage or moiety, and wherein position 1 is the first nucleoside starting from the 5′-terminus of the nucleic acid molecule.
2. The nucleic acid molecule of claim 1 , wherein at least two or three of positions 24, 25 and 26 comprise a phosphorothioate linkage or moiety.
3. The nucleic acid molecule of claim 1 , wherein at least one of positions 1, 6, 9, 12, 15, 17, 18, 22, 28 and 29 of the nucleic acid molecule comprise a 2′-O-methyl modification.
4. The nucleic acid molecule of claim 1 , wherein at least two, three, four, five, six, seven, eight, nine or ten of positions 1, 6, 9, 12, 15, 17, 18, 22, 28 and 29 of the nucleic acid molecule comprise a 2′-O-methyl modification.
5. The nucleic acid molecule of claim 1 , wherein the C-5 modified pyrimidine independently, and for each occurrence, is selected from the group consisting a 5-(N-benzylcarboxyamide)-2′-deoxyuridine (BndU), 5-(N-benzylcarboxyamide)-2′-O-methyluridine, 5-(N-benzylcarboxyamide)-2′-fluorouridine, 5-(N-phenethylcarboxyamide)-2′-deoxyuridine (PedU), 5-(N-thiophenylmethylcarboxyamide)-2′-deoxyuridine (ThdU), 5-(N-isobutylcarboxyamide)-2′-deoxyuridine (iBudU), 5-(N-isobutylcarboxyamide)-2′-O-methyluridine, 5-(N-isobutylcarboxyamide)-2′-fluorouridine, 5-(N-tryptaminocarboxyamide)-2′-deoxyuridine (TrpdU), 5-(N-tryptaminocarboxyamide)-2′-O-methyluridine, 5-(N-tryptaminocarboxyamide)-2′-fluorouridine, 5-(N-[1-(3-trimethylamonium) propyl]carboxyamide)-2′-deoxyuridine chloride, 5-(N-naphthylmethylcarboxyamide)-2′-deoxyuridine (NapdU), 5-(N-naphthylmethylcarboxyamide)-2′-O-methyluridine, 5-(N-2-naphthylmethylcarboxyamide)-2′-deoxyuridine (2NapdU), 5-(N-2-naphthylmethylcarboxyamide)-2′-O-methyluridine, 5-(N-naphthylmethylcarboxyamide)-2′-fluorouridine, and 5-(N-[1-(2,3-dihydroxypropyl)]carboxyamide)-2′-deoxyuridine).
6. The nucleic acid molecule of claim 1 , wherein the C-5 modified pyrimidine is a 5-(N-naphthylmethylcarboxyamide)-2′-deoxyuridine (NapdU).
7. The nucleic acid molecule of claim 1 , wherein the nucleic acid molecule is selected from the group consisting of SEQ ID NOS: 706 and 763-799.
8. The nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises nine or ten 2′-O-methylnucleosides and one, two or three phosphorothioate linkages or moieties.