PDGF and VEGF aptamers having improved stability and their use in treating PDGF and VEGF mediated diseases and disorders

• US 9,695,424 B2
• Filed: 09/08/2014
• Issued: 07/04/2017
• Est. Priority Date: 09/09/2013
• Status: Active Grant

• Current Assignee: SomaLogic Operating Co., Inc. (Somalogic Incorporated)
• Original Assignee: Somalogic Incorporated
• Inventors: Drolet, Daniel W., Gelinas, Amy D., Zhang, Chi, Vrkljan, Michael, Janjic, Nebojsa
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US14/909,305
• Publication Number:
  US 20160177307A1
• Time in Patent Office: 1,030 Days
• Field of Search: 536 231, 514 44 R
• 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 27/02 Ophthalmic agents
• 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 7/10 Antioedematous agents; Diuretics
• 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: 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 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance

Abstract

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.

Claims

1. A nucleic acid molecule comprising the nucleic sequence

5′

-C-G-A-C-A-G-C-A-Z-G-Z-A-Z-G-C-A-C-A-Z-C-Z-3′

(SEQ ID NO;

     830),

wherein z is a C-5 modified pyrimidine, and at least one of positions 4, 9, 10, 11 and 12 of the nucleic acid sequence comprise a phosphorothioate linkage or moiety, wherein position 1 is the first nucleoside starting from the 5′

-terminus of the nucleic acid sequence.

2. The nucleic acid molecule of claim 1, wherein at least two, three, four or five of positions 4, 9, 10, 11 and 12 of the nucleic acid sequence comprise a phosphorothioate linkage or moiety, and wherein position 1 is the first nucleoside starting from the 5′

-terminus of the nucleic acid sequence.

3. The nucleic acid molecule of claim 1, wherein at least one of positions 1, 4, 6, 7, 14, 15, 16, 17, 18, 20 and 21 of the nucleic acid sequence comprise a 2′

-O-methyl modification, and wherein position 1 is the first nucleoside starting from the 5′

-terminus of the nucleic acid sequence.

4. The nucleic acid molecule of claim 1, wherein at least two, three, four, five, six, seven, eight, nine, ten or eleven of positions 1, 4, 6, 7, 14, 15, 16, 17, 18, 20 and 21 of the nucleic acid sequence comprise a 2′

-O-methyl modification, and wherein position 1 is the first nucleoside starting from the 5′

-terminus of the nucleic acid sequence.

5. The nucleic acid molecule of claim 1, wherein the nucleic acid molecule is selected from the group consisting of SEQ ID NOS:

545 and 800-821.

6. The nucleic acid molecule of claim 1, wherein the nucleic acid molecule comprises eleven 2′

-O-methyl nucleosides and four or five phosphorothioate linkages or moieties.

7. 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).

8. The nucleic acid molecule of claim 1, wherein the C-5 modified pyrimidine is a 5-(N-naphthylmethylcarboxyamide)-2′

-deoxyuridine (NapdU).

9. The nucleic acid molecule of claim 3, wherein Z is a 5-(N-naphthylmethylcarboxyamide)-2′

-deoxyuridine (NapdU).