Nucleic acid compounds for binding growth differentiation factor 8

1. An aptamer that binds GDF8, wherein the aptamer comprises the sequence:

5′-GWWR 1 C n R 2 ZG n GWC n X 2 CW n WWX 1 A n R 1 C n X 1 KW n AGX 2 n WCX 1 ZD-3′ (SEQ ID NO: 1),

wherein:

each W is independently, and for each occurrence, a C-5 modified pyrimidine;

X 1 and X 2 are each independently selected from W and C;

R 1 and R 2 are each independently selected from G and A;

each Z is independently selected from A and W;

K is selected from G and W;

D is selected from G, A, and W; and

each n is independently selected from 0 and 1;

or wherein the aptamer comprises the sequence:

5′-X 1 GWWR 1 C n R 2 ZG n GWC n X 2 CW n WWX 1 A n R 1 C n X 1 KW n AGX 2 n WCX 1 ZD-3′ (SEQ ID NO: 2),

wherein:

each W is independently, and for each occurrence, a C-5 modified pyrimidine;

X 1 and X 2 are each independently selected from W and C;

R 1 and R 2 are each independently selected from G and A;

each Z is independently selected from A and W;

K is selected from G and W;

D is selected from G, A, and W; and

each n is independently selected from 0 and 1;

or wherein the aptamer comprises the sequence:

5′-GWWR 1 R 2 ZGWC n X 2 CW n WWX 1 A n R 1 C n X 1 KAGWCX 1 ZD-3′ (SEQ ID NO: 49), wherein:

each W is independently, and for each occurrence, a C-5 modified pyrimidine;

X 1 and X 2 are each independently selected from W and C;

R 1 and R 2 are each independently selected from G and A;

each Z is independently selected from A and W;

K is selected from G and W;

D is selected from G, A, and W; and

each n is independently selected from 0 and 1;

or wherein the aptamer comprises the sequence:

5′-WGWWAGZGWX 2 CWWX 1 AWGAGWCWAD-3′ (SEQ ID NO: 3), wherein:

each W is independently, and for each occurrence, a C-5 modified pyrimidine;

X 1 and X 2 are each independently selected from W and C;

Z is selected from A and W; and

D is selected from G, A, and W;

or wherein the aptamer comprises the sequence:

5′-GWWAGAGWCCWWWAWGAGWCWAG-3′ (SEQ ID NO: 4), wherein each W is independently, and for each occurrence, a C-5 modified pyrimidine.

2. The aptamer of claim 1 , wherein the aptamer binds GDF8 with an affinity of less than 10 nM, wherein, under the same conditions, the aptamer binds GDF11 with an affinity that is at least 10-fold weaker than the affinity for GDF8 or does not bind GDF 11.

3. The aptamer of claim 2 , wherein the aptamer binds GDF8 with an affinity of less than 8 nM and binds GDF11 with an affinity greater than 100 nM.

4. The aptamer of claim 2 , wherein affinity is determined using a binding assay comprising a polyanionic inhibitor selected from dextran sulfate, heparin, Z-block, poly-dl/dC, sonicated or sheared salmon sperm DNA, calf thymus DNA, and dNTPs.

5. The aptamer of claim 1 , wherein at least one R 1 is A.

6. The aptamer of claim 1 , wherein at least one R 2 is G.

7. The aptamer of claim 1 , wherein K is G.

8. The aptamer of claim 1 , wherein at least 1 n is 0.

9. The aptamer of claim 1 , wherein at least one X 1 is W.

10. The aptamer of claim 1 , wherein at least one X 2 is C.

11. The aptamer of claim 1 , wherein each W is independently, and for each occurrence, 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-tyrosylcarboxyamide)-2′-deoxyuridine (TyrdU),

5-(N-3,4-methylenedioxybenzylcarboxyamide)-2′-deoxyuridine (MBndU),

5-(N-4-fluorobenzylcarboxyamide)-2′-deoxyuridine (FBndU),

5-(N-3-phenylpropylcarboxyamide)-2′-deoxyuridine (PPdU),

5-(N-imidizolylethylcarboxyamide)-2′-deoxyuridine (ImdU),

5-(N-isobutylcarboxyamide)-2′-O-methyluridine,

5-(N-isobutylcarboxyamide)-2′-fluorouridine,

5-(N-tryptaminocarboxyamide)-2′-deoxyuridine (TrpdU),

5-(N-R-threoninylcarboxyamide)-2′-deoxyuridine (ThrdU),

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,

5-(N-[1-(2,3-dihydroxypropyl)]carboxyamide)-2′-deoxyuridine),

5-(N-2-naphthylmethylcarboxyamide)-2′-deoxyuridine (2NapdU),

5-(N-2-naphthylmethylcarboxyamide)-2′-O-methyluridine,

5-(N-2-naphthylmethylcarboxyamide)-2′-fluorouridine,

5-(N-1-naphthylethylcarboxyamide)-2′-deoxyuridine (NEdU),

5-(N-1-naphthylethylcarboxyamide)-2′-O-methyluridine,

5-(N-1-naphthylethylcarboxyamide)-2′-fluorouridine,

5-(N-2-naphthylethylcarboxyamide)-2′-deoxyuridine (2NEdU),

5-(N-2-naphthylethylcarboxyamide)-2′-O-methyluridine,

5-(N-2-naphthylethylcarboxyamide)-2′-fluorouridine,

5-(N-3-benzofuranylethylcarboxyamide)-2′-deoxyuridine (BF dU),

5-(N-3-benzofuranylethylcarboxyamide)-2′-O-methyluridine,

5-(N-3-benzofuranylethylcarboxyamide)-2′-fluorouridine,

5-(N-3-benzothiophenylethylcarboxyamide)-2′-deoxyuridine (BTdU),

5-(N-3-benzothiophenylethylcarboxyamide)-2′-O-methyluridine, and

5-(N-3-benzothiophenylethylcarboxyamide)-2′-fluorouridine.

12. The aptamer of claim 1 , wherein at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6 W are each independently selected from 5-(N-tryptaminocarboxyamide)-2′-deoxyuridine (TrpdU), 5-(N-tryptaminocarboxyamide)-2′-O-methyluridine, and 5-(N-tryptaminocarboxyamide)-2′-fluorouridine.

13. The aptamer of claim 12 , wherein each W is independently selected from 5-(N-tryptaminocarboxyamide)-2′-deoxyuridine (TrpdU), 5-(N-tryptaminocarboxyamide)-2′-O-methyluridine, and 5-(N-tryptaminocarboxyamide)-2′-fluorouridine.

14. The aptamer of claim 1 , wherein the aptamer consists of 18 to 200 nucleotides, wherein each nucleotide may, independently, be a modified or unmodified nucleotide.

15. The aptamer of claim 1 , wherein the aptamer comprises a detectable label.

16. The aptamer of claim 2 , wherein GDF8 is mature human GDF8 comprising the sequence of SEQ ID NO: 63 and GDF11 is mature human GDF11 comprising the sequence of SEQ ID NO: 50.

17. A method of detecting GDF8 in a sample, comprising contacting proteins from the sample with an aptamer of claim 1 .

18. An aptamer that binds GDF 8, wherein the aptamer comprises a sequence selected from SEQ ID NOs: 5 to 48.

19. The aptamer of claim 1 , wherein each R 1 is A.

20. The aptamer of claim 1 , wherein each R 2 is G.

21. The aptamer of claim 1 , wherein each n is 0.

22. The aptamer of claim 1 , wherein each X 1 is W.

23. The aptamer of claim 1 , wherein each X 2 is C.