Aptamers to PDGF and VEGF and their use in treating PDGF and VEGF mediated conditions
Aptamers 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. An aptamer comprising the sequence:
(SEQ ID NO. 512)
5′-ACAL n ZGZAZGL m ZLZ-3′
wherein;
each Z is, independently, a modified pyrimidine;
each L is independently selected from a substituted or unsubstituted C 2 -C 50 linker, a polyethylene glycol linker, and a modified or unmodified nucleotide;
at least one, at least two, at least three, at least four, or at least five nucleosides comprise a 2′-OMe;
n is 1 to 5; and
m is 1 to 10.
2. The aptamer of claim 1 , wherein each Z is independently selected from 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-naphthylmethylcarboxyamide)-2′-fluorouridine, and 5-(N-[1-(2,3-dihydroxypropyl)]carboxyamide)-2′-deoxyuridine).
3. The aptamer of claim 1 , wherein n is 1, 2, 3, or 4; and wherein m is 1, 2, 3, 4, 5, 6, 7, 8, or 9.
4. The aptamer of 1 further comprising at least one, at least two, at least three, at least four, or at least five internucleoside linkages are phosphorothioate linkages.
5. The aptamer of 1 , wherein the aptamer binds PDGF with an affinity of less than 10 nM, less than 5 nM, less than 2 nM, or less than 1 nM.
6. The aptamer of claim 1 , wherein the aptamer inhibits PDGF-mediated phosphorylation of a PDGF receptor.
7. The aptamer of claim 1 , wherein L is independently, for each occurrence, selected from the group consisting of a substituted or unsubstituted C 2 -C 10 linker, a substituted or unsubstituted C 2 -C 8 linker, a substituted or unsubstituted C 2 -C 6 linker, a substituted or unsubstituted C 2 -C 5 linker, a substituted or unsubstituted C 2 -C 4 linker, a substituted or unsubstituted C 3 linker, a G, a C and an A.
8. A method comprising administering to a subject in need a therapeutically effective amount of the aptamer of claim 1 , wherein the method treats and/or prevents a conditions selected from the group consisting of macular degeneration, an ophthalmic condition, fibrosis, a cardiovascular disease and a cancer.
9. An aptamer comprising the sequence:
(SEQ ID NO. 512)
5′-ACAL n ZGZAZGL m ZLZ-3′
wherein;
each Z is, independently, selected from the group consisting of 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-naphthylmethylcarboxyamide)-2′-fluorouridine, and 5-(N-[1-(2,3-dihydroxypropyl)]carboxyamide)-2′-deoxyuridine);
each L is independently selected from a substituted or unsubstituted C 2 -C 50 linker, a polyethylene glycol linker, and a modified or unmodified nucleotide;
n is 1 to 5; and
m is 1 to 10.
10. The aptamer of claim 9 further comprising at least one, at least two, at least three, at least four, or at least five nucleosides comprise a 2′-OMe.
11. The aptamer of claim 9 , wherein n is 1, 2, 3, or 4; and wherein m is 1, 2, 3, 4, 5, 6, 7, 8, or 9.
12. The aptamer of 9 further comprising at least one, at least two, at least three, at least four, or at least five internucleoside linkages are phosphorothioate linkages.
13. The aptamer of 9 , wherein the aptamer binds PDGF with an affinity of less than 10 nM, less than 5 nM, less than 2 nM, or less than 1 nM.
14. The aptamer of claim 9 , wherein the aptamer inhibits PDGF-mediated phosphorylation of a PDGF receptor.
15. The aptamer of claim 9 , wherein L is independently, for each occurrence, selected from the group consisting of a substituted or unsubstituted C 2 -C 10 linker, a substituted or unsubstituted C 2 -C 8 linker, a substituted or unsubstituted C 2 -C 6 linker, a substituted or unsubstituted C 2 -C 5 linker, a substituted or unsubstituted C 2 -C 4 linker, a substituted or unsubstituted C 3 linker, a G, a C and an A.
16. A method comprising administering to a subject in need a therapeutically effective amount of the aptamer of claim 9 , wherein the method treats and/or prevents a conditions selected from the group consisting of macular degeneration, an ophthalmic condition, fibrosis, a cardiovascular disease and a cancer.
17. An aptamer comprising the sequence:
(SEQ ID NO. 512)
5′-ACAL n ZGZAZGL m ZLZ-3′
wherein;
each Z is, independently, a modified pyrimidine;
each L is independently, for each occurrence, selected from the group consisting of a substituted or unsubstituted C 2 -C 10 linker, a substituted or unsubstituted C 2 -C 8 linker, a substituted or unsubstituted C 2 -C 6 linker, a substituted or unsubstituted C 2 -C 5 linker, a substituted or unsubstituted C 2 -C 4 linker, a substituted or unsubstituted C 3 linker, a G, a C and an A;
n is 1 to 5; and
m is 1 to 10.
18. The aptamer of claim 17 , wherein each Z is independently selected from 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-naphthylmethylcarboxyamide)-2′-fluorouridine, and 5-(N-[1-(2,3-dihydroxypropyl)]carboxyamide)-2′-deoxyuridine).
19. The aptamer of claim 17 , wherein n is 1, 2, 3, or 4; and wherein m is 1, 2, 3, 4, 5, 6, 7, 8, or 9.
20. The aptamer of claim 17 further comprising at least one, at least two, at least three, at least four, or at least five nucleosides comprise a 2′-OMe.
21. The aptamer of 17 further comprising at least one, at least two, at least three, at least four, or at least five internucleoside linkages are phosphorothioate linkages.
22. The aptamer of 17 , wherein the aptamer binds PDGF with an affinity of less than 10 nM, less than 5 nM, less than 2 nM, or less than 1 nM.
23. The aptamer of claim 17 , wherein the aptamer inhibits PDGF-mediated phosphorylation of a PDGF receptor.
24. A method comprising administering to a subject in need a therapeutically effective amount of the aptamer of claim 17 , wherein the method treats and/or prevents a conditions selected from the group consisting of macular degeneration, an ophthalmic condition, fibrosis, a cardiovascular disease and a cancer.