Aptamers to PDGF and VEGF and their use in treating PDGF and VEGF mediated conditions

• US 9,410,156 B2
• Filed: 03/28/2013
• Issued: 08/09/2016
• Est. Priority Date: 03/28/2012
• Status: Active Grant

• Current Assignee: SomaLogic Operating Co., Inc. (Somalogic Incorporated)
• Original Assignee: Somalogic Incorporated
• Inventors: Jarvis, Thale C., Rohloff, John C., Gelinas, Amy D., Zhang, Chi, Drolet, Daniel W., Waugh, Sheela M., Janjic, Nebojsa
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US14/381,679
• Publication Number:
  US 20150105452A1
• Time in Patent Office: 1,230 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/04 Artificial tears; Irrigation solutions
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 27/12 for cataracts
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/16 Aptamers
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/318 where the PO2 is completely replaced, e.g. MMI or formacetal
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2310/3533 : Halogen
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications

Abstract

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.

Claims

1. An aptamer comprising the sequence:

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.

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 C₂-C₁₀ linker, a substituted or unsubstituted C₂-C₈ linker, a substituted or unsubstituted C₂-C₆ linker, a substituted or unsubstituted C₂-C₅ linker, a substituted or unsubstituted C₂-C₄ linker, a substituted or unsubstituted C₃ 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.

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.

9. An aptamer comprising the sequence:

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.

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 C₂-C₁₀ linker, a substituted or unsubstituted C₂-C₈ linker, a substituted or unsubstituted C₂-C₆ linker, a substituted or unsubstituted C₂-C₅ linker, a substituted or unsubstituted C₂-C₄ linker, a substituted or unsubstituted C₃ 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.

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.

17. An aptamer comprising the sequence:

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.

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.

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.