Von Willebrand Factor (VWF)—targeting agents and methods of using the same

1. An aptamer comprising

a polynucleotide comprising from 5′ to 3′ (a) a polynucleotide having at least 70% sequence identity to SEQ ID NO: 1 comprising a first stem forming region comprising 3 nucleotides, a first loop region comprising the nucleotide sequence AAC, a second stem forming region comprising 3 nucleotides, a second loop region comprising the nucleotide sequence CC and a third stem forming region consisting of 2-8 nucleotides, (b) a third loop region consisting of 1-12 nucleotides or a spacer sequence, and (c) a polynucleotide having at least 70% sequence identity to SEQ ID NO:2 comprising a fourth stem forming region consisting of 2-8 nucleotides and forming a stem with the third stem forming region, a fourth loop region comprising the nucleotide C, a fifth stem forming region comprising 3 nucleotides and forming a stem with the second stem forming region, a fifth loop region comprising the nucleotide sequence CAGA, and a sixth stem forming region comprising 3 nucleotides and forming a stem with the first stem forming region,

wherein the polynucleotide comprises an unmodified form or comprises a modified form comprising at least one nucleotide base modification, and

wherein the aptamer is no more than 53 nucleotides in length.

2. The aptamer of claim 1 , wherein the aptamer comprises from 5′ to 3′ SEQ ID NO: 1, a variable nucleotide sequence consisting of 1-12 nucleotides or a spacer sequence, and SEQ ID NO: 2.

3. The aptamer of claim 1 , wherein the aptamer comprises a polynucleotide having at least 90% sequence identity to any one of SEQ ID NOS: 3 or 4.

4. The aptamer of claim 1 , wherein the dissociation constant (K D ) of the aptamer for the human VWF protein is less than 100 nanomolar (nM).

5. The aptamer of claim 1 , further comprising a tail nucleotide sequence at the 5′ end or the 3′ end of the polynucleotide which is not capable of base pairing with 3 or more consecutive nucleotides in the polynucleotide, wherein the tail nucleotide sequence consists of 2-12 nucleotides.

6. The aptamer of claim 5 , wherein the tail nucleotide sequence is at the 3′ end of the polynucleotide and consists of the nucleotide sequence (U/T)(U/T)(U/T)(U/T)(U/T).

7. The aptamer of claim 1 , wherein the aptamer is no more than 39 nucleotides in length.

8. The aptamer of claim 1 , wherein the polynucleotide comprises a modified form comprising at least one nucleotide base modification selected from the group consisting of a 2′fluoro modification, a 2′O-methyl modification, a 5′ modification, and a 3′modification.

9. The aptamer of claim 1 , wherein the aptamer comprises SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.

10. The aptamer of claim 1 , wherein the polynucleotide comprises a 5′ linker and/or a 3′ linker.

11. The aptamer of claim 1 , wherein the polynucleotide further comprises a stability agent.

12. The aptamer of claim 11 , wherein the stability agent is selected from the group consisting of polyethylene glycol (PEG), cholesterol, albumin, and Elastin-like polypeptide.

13. The aptamer of claim 11 , wherein the polynucleotide and the stability agent are linked by a covalent bond or via a tag system.

14. A dimer, trimer, or tetramer comprising the aptamers of claim 1 .

15. A pharmaceutical composition comprising a pharmaceutical carrier and the composition of claim 1 .

16. A method for preventing blood clot formation in a subject comprising administering to the subject the composition of claim 1 in a therapeutically effective amount to prevent blood clot formation in the subject.

17. The method of claim 16 , wherein the subject suffers from atrial fibrillation or is at risk of having a Deep Vein Thrombosis, a stroke, a heart attack, or a pulmonary embolism.

18. The method of claim 16 , further comprising administering to the subject an antidote in a therapeutically effective amount to neutralize the aptamer or the VWF-targeting agent.

19. The method of claim 16 , wherein the subject is a mammal.