Nucleic acids specifically binding CGRP
The present invention is related to a nucleic acid molecule capable of binding to CGRP, wherein the nucleic acid molecule comprises a central stretch of nucleotides, wherein the central stretch of nucleotides comprises a nucleotide sequence of 5′ HWn 1 n 2 YGGAn 3 An 4 UMn 5 n 6 Yn 7 n 8 n 9 n 10 n 11 Kn 12 Rn 13 ADn 14 n 15 ARn 16 Un 17 Cn 18 n 19 Un 20 n 21 3′ [SEQ ID NO: 99], wherein H, W, Y, G, A, U, M, B, K, R, D, C are ribonucleotides, and n 1 is R or dG, n 2 is U or dT, n 3 is K or dG, n 4 is C or dC, n 5 is M or dC, n 6 is B or dU, n 7 is N or dG, n 8 is Y or dT, n 9 is N or dC, n 10 is R or dG, n 11 is V or dA, n 12 is K or dT or dU, n 13 is G or dG, n 14 is A or dA, n 15 is U or dT, n 16 is R or dG, n 17 is Y or dC, n 18 is C or dC, n 19 is B or dC, n 20 is C or dC, n 21 is C or dC, and dG, dT, dC, dA and dU are 2′-deoxyribonucleotides.
1. An L-nucleic acid that binds CGRP, or a homolog of said L-nucleic acid, wherein said L-nucleic acid comprises SEQ ID NO:45, and the homolog has at least 85% homology to SEQ ID NO:45.
2. The L-nuc eic acid according to claim 1 , wherein the L-nucleic acid is an antagonist of an activity mediated by CGRP.
3. The L-nucleic acid according to claim 1 , wherein the CGRP is human CGRP, mouse CGRP, rat CGRP or CGRP from Maca mulatta.
4. The L-nucleic acid according to claim 1 , wherein the CGRP is human CGRP.
5. The L-nucleic acid according to claim 1 , wherein the CGRP is α-CGRP or β-CGRP.
6. The L-nucleic acid according to claim 1 , wherein the CGRP is a human α-CGRP or a rat α-CGRP.
7. The L-nucleic acid according tea claim 1 , wherein the L-nucleic acid has binding affinity to human α-CGRP, expressed as K D , of 10 nM or below.
8. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid has binding affinity to human α-CGRP, expressed as K D , of 1 nM or below.
9. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid has binding affinity to human α-CGRP, expressed as IC50, of 10 nM or below.
10. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid has binding affinity to human α-CGRP, expressed as IC50, of 1 nM or below.
11. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid has binding affinity to human amylin, expressed as K D , of 100 nM or more.
12. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid has binding affinity to human amylin, expressed as IC50, of 100 nM or more.
13. The L-nucleic acid according to claim 1 , wherein
the L-nucleic acid has binding affinity to human α-CGRP, expressed as K D , of 10 nM or below, and to human amylin of 100 nM or more, or
the L-nucleic acid has binding affinity to human α-CGRP, expressed as IC50, of 10 nM or below, and to human amylin of 100 nM or more.
14. The L-nucleic acid according to claim 1 , wherein
the L-nucleic acid has binding affinity to human α-CGRP, expressed as K D , of 1 nM or below, and to human amylin of 100 nM or more, or
the L-nucleic acid has binding affinity to human α-CGRP, expressed as IC50, 1 nk or below, and to human amylin of 100 nM or more.
15. The L-nucleic acid according to claim 1 , wherein the L-nucleic acid comprises a modification group.
16. The L-nucleic acid according to claim 15 , wherein the modification group is selected from the group consisting of polyethylene glycol, linear polyethylene glycol, branched polyethylene glycol, hydroxyethyl starch, a peptide, a protein, a polysaccharide, polyoxypropylene, polyoxyamidate, poly (2-hydroxyethyl)-L-glutamine and a sterol.
17. The L-nucleic acid according to claim 16 , wherein the linear or branched polyethylene glycol comprises a molecular weight of from about 20,000 to about 120,000 Da.
18. The L-nucleic acid according to claim 15 , wherein the hydroxyethyl starch comprises a molecular weight of from about 100 to about 700 kDa.
19. The L-nucleic acid according to claim 15 , wherein the modification group is coupled to the L-nucleic acid by a linker.
20. The L-nucleic acid according to claim 15 , wherein the modification group is at a terminus of the L-nucleic acid.
21. The L-nucleic acid according to claim 15 , wherein the modification group is at the 5′ terminus of the L-nucleic acid.
22. A pharmaceutical composition comprising the L-nucleic acid according to claim 1 and a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier, a pharmaceutically active agent or a combination thereof.
23. A complex comprising the L-nucleic acid according to claim 1 and CGRP.
24. A method for the detection of the L-nucleic acid according to claim 1 in a sample, wherein the method comprises the steps of:
a) providing a capture probe, wherein the capture probe is at least partially complementary to a first part of the L-nucleic acid according to claim 1 , and a detection probe, wherein the detection probe is at least partially complementary to a second part of the L-nucleic acid according to claim 1 , or, alternatively, the capture probe is at least partially complementary to a second part of the L-nucleic acid according to claim 1 and the detection probe is at least partially complementary to the first part of the L-nucleic acid according to claim 1 ;
b) adding the capture probe and the detection probe separately or combined to a sample containing the L-nucleic acid according to claim 1 or presumed to contain the L-nucleic acid according to claim 1 ;
c) allowing the capture probe and the detection probe to react either simultaneously or in any order sequentially with the L-nucleic acid according to claim 1 or part thereof;
d) optionally detecting whether or not the capture probe is hybridized to the L-nucleic acid according to claim 1 provided in step a); and
e) detecting a complex formed in step c) consisting of the L-nucleic acid according to claim 1 and the capture probe and the detection probe.