Aptamers to β-NGF and their use in treating β-NGF mediated diseases and disorders
The present disclosure relates generally to the field of nucleic acids and, more particularly, to aptamers capable of binding to β-NGF; pharmaceutical compositions comprising such β-NGF aptamers; and methods of making and using the same.
1. An aptamer comprised of the sequence
(SEQ ID NO: 154)
BAZGRGGRS N (0-1) ZWGGGG N (0-1) ZZWADCCGZZRZG
wherein
B is selected from a C, G or Z;
R is independently selected from an A or G;
S is selected from a C or G;
W is independently selected from a Z or T;
D is selected from an A, G or Z;
N is independently selected from any naturally occurring or modified nucleotide and
Z is independently selected from a modified pyrimidine.
2. The aptamer of claim 1 , wherein the aptamer binds to β-NGF.
3. The aptamer of claim 2 , wherein the aptamer inhibits the function of β-NGF.
4. The aptamer of claim 2 , wherein said aptamer has the ability to modulate the binding of β-NGF to its one or more of its cellular receptors.
5. The aptamer of claim 4 wherein said cellular receptor is selected from p75 or TrkA.
6. The aptamer of claim 2 , wherein the aptamer comprises a K d for β-NGF of 30 nM or less.
7. The aptamer of claim 1 , wherein said modified pyrimidine is a C-5 modified pyrimidine.
8. The aptamer of claim 7 , wherein the C-5 modified pyrimidine is selected from the group consisting of the following compounds:
wherein
R″″ is selected from the group consisting of a branched or linear lower alkyl (C1-C20); halogen (F, Cl, Br, I); nitrile (CN); boronic acid (BO 2 H 2 ); carboxylic acid (COOH); carboxylic acid ester (COOR″); primary amide (CONH 2 ); secondary amide (CONHR″); tertiary amide (CONR″R′″); sulfonamide (SO 2 NH 2 ); N-alkylsulfonamide (SONHR″);
wherein
R″, R′″ are independently selected from a group consisting of a branched or linear lower alkyl (C1-C2)); phenyl (C 6 H 5 ); an R″″ substituted phenyl ring (R″″C 6 H 4 ); wherein R″″ is defined above; a carboxylic acid (COOH); a carboxylic acid ester (COOR′″″); wherein R′″″ is a branched or linear lower alkyl (C1-C20); and cycloalkyl; wherein R″═R′″═(CH 2 )n; and
wherein n=2-10.
9. The aptamer of claim 7 , wherein the C-5 modified pyrimidine is selected from the group consisting of 5-(N-benzylcarboxyamide)-2′-deoxyuridine (BndU), 5-(N-isobutylcarboxyamide)-2′-deoxyuridine (iBudU), 5-(N-tryptaminocarboxyamide)-2′-deoxyuridine (TrpdU) and 5-(N-naphthylmethylcarboxyamide)-2′-deoxyuridine (NapdU).
10. The aptamer of claim 7 , wherein the C-5 modified pyrimidine is 5-(N-benzylcarboxyamide)-2′-deoxyuridine (BndU).
11. The aptamer of claim 1 , wherein said aptamer has the following sequence:
(SEQ ID NO: 45)
BAZGRGGRSZWGGGGZZWADCCGZZRZG.
12. An aptamer comprised of the sequence
(SEQ. ID. NO: 3)
BAZGRGGRSZZGGGGZZZADCCGZZRZG,
wherein
B is selected from a C, G or Z;
R is independently selected from an A or G;
S is selected from a C or G;
D is selected from an A, G or Z; and
Z is independently selected from a modified pyrimidine.
13. An aptamer that binds to β-NGF comprising a sequence selected from the group consisting of SEQ. ID. NOS: 1, 2, 9-44 and 149.
14. The aptamer of claim 13 , wherein the aptamer inhibits the function of β-NGF.
15. The aptamer of claim 13 , wherein said aptamer has the ability to modulate the binding of β-NGF to its one or more of its cellular receptors.
16. The aptamer of claim 15 , wherein said cellular receptor is selected from p75 or TrkA.
17. The aptamer of claim 16 , wherein the sequence has at least about 90% identity, at least about 91% identity, at least about 92% identity, at least about 93% identity, at least about 94% identity, or at least about 95% identity.