Methods and compositions for 18F-radiolabeling of biologics
The invention relates to water soluble 18 F-prosthetic groups and the synthesis and use of 18 F-labeled biological molecules containing the 18 F-prosthetic groups for imaging various processes within the body, for detecting the location of molecules associated with disease pathology, and for monitoring disease progression are disclosed.
1. An 18 F-radiolabeled protein-based probe comprising 18 F-radiolabeled prosthetic group, a bifunctional conjugating (BFC) moiety and a protein with the following structure,
wherein the 18 F is ortho to the N atom, x is an integer from 1 to 8, and the BFC comprises a cyclooctyne moiety and a reactive group that forms a covalent bond with an amine, carboxyl, carbonyl or thiol functional group on the protein, or pharmaceutically acceptable salt thereof.
2. The 18 F-radiolabeled protein-based probe of claim 1 , wherein x is 4.
3. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the BFC further comprises a polyethylene glycol (PEG) y spacer arm, wherein y is an integer from 1 to 8.
4. The 18 F-radiolabeled protein-based probe of claim 3 , wherein the BFC is DBCO-PEG4-NHS-Ester, DBCO-PEG4-Acid, DBCO-PEG4-Amine or DBCO-PEG4-Maleimide.
5. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the protein portion of the probe comprises a fibronectin type III (Fn3) domain.
6. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the protein portion of the probe comprises an antibody or antibody fragment.
7. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the [O(CH 2 ) 2 ] x moiety is present in the 1-3 configuration relative to the nitrogen on the pyridine ring.
8. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the [O(CH 2 ) 2 ] x moiety is present in the 1-2 configuration relative to the nitrogen on the pyridine ring.
9. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the [O(CH 2 ) 2 ] x moiety is present in the 1-4 configuration relative to the nitrogen on the pyridine ring.
10. The 18 F-radiolabeled protein-based probe of claim 1 , wherein the BFC is covalently linked to a thiol group on a cysteine residue of the protein.
11. The 18 F-radiolabeled protein-based probe of claim 10 , wherein the cysteine residue is at the C-terminus of the protein.
12. The 18 F-radiolabeled protein-based probe of claim 1 , having the structure
13. The 18 F-radiolabeled protein-based probe of claim 12 , wherein the BFC further comprises a polyethylene glycol (PEG) y spacer arm, wherein y is an integer from 1 to 8.
14. The 18 F-radiolabeled protein-based probe of claim 13 , wherein the BFC is DBCO-PEG4-NHS-Ester, DBCO-PEG4-Acid, DBCO-PEG4-Amine or DBCO-PEG4-Maleimide.
15. The 18 F-radiolabeled protein-based probe of claim 14 , wherein the BFC is DBCO-PEG4-Maleimide.
16. The 18 F-radiolabeled protein-based probe of claim 12 , wherein the protein portion of the probe comprises a fibronectin type III (Fn3) domain.
17. The 18 F-radiolabeled protein-based probe of claim 12 , wherein the BFC is covalently linked to a thiol group on a cysteine residue of the protein.
18. The 18 F-radiolabeled protein-based probe of claim 17 , wherein the cysteine residue is at the C-terminus of the protein.
19. The 18 F-radiolabeled protein-based probe of claim 12 , wherein the protein portion of the probe comprises an antibody or antibody fragment.
20. The 18 F-radiolabeled protein-based probe of claim 1 , having the following structure,
wherein the maleimide group of the BFC is covalently linked to a thiol group on a cysteine residue of the protein.
21. The 18 F-radiolabeled protein-based probe of claim 20 , wherein the cysteine residue is at the C-terminus of the protein.
22. The 18 F-radiolabeled protein-based probe of claim 20 , wherein the protein portion of the probe comprises a fibronectin type III (Fn3) domain.
23. The 18 F-radiolabeled protein-based probe of claim 20 , wherein the protein portion of the probe comprises an antibody or antibody fragment.
24. The 18 F-radiolabeled protein-based probe of claim 1 , having the following structure,
25. The 18 F-radiolabeled protein-based probe of claim 24 , wherein the protein portion of the probe comprises a fibronectin type III (Fn3) domain, an antibody or an antibody fragment.
26. The 18 F-radiolabeled protein-based probe of claim 24 , wherein the BFC is covalently linked to a thiol group on a cysteine residue of the protein.
27. The 18 F-radiolabeled protein-based probe of claim 26 , wherein the cysteine residue is at the C-terminus of the protein.
28. The 18 F-radiolabeled protein-based probe of claim 24 , wherein the BFC further comprises a polyethylene glycol (PEG) y spacer arm, wherein y is an integer from 1 to 8.
29. The 18 F-radiolabeled protein-based probe of claim 28 , wherein the BFC is DBCO-PEG4-NHS-Ester, DBCO-PEG4-Acid, DBCO-PEG4-Amine or DBCO-PEG4-Maleimide.
30. The 18 F-radiolabeled protein-based probe of claim 1 , having the following structure,
31. The 18 F-radiolabeled protein-based probe of claim 30 , wherein the protein portion of the probe comprises a fibronectin type III (Fn3) domain, an antibody or an antibody fragment.
32. The 18 F-radiolabeled protein-based probe of claim 30 , wherein the BFC is covalently linked to a thiol group on a cysteine residue of the protein.
33. The 18 F-radiolabeled protein-based probe of claim 32 , wherein the cysteine residue is at the C-terminus of the protein.
34. The 18 F-radiolabeled protein-based probe of claim 30 , wherein the BFC further comprises a polyethylene glycol (PEG) y spacer arm, wherein y is an integer from 1 to 8.
35. The 18 F-radiolabeled protein-based probe of claim 34 , wherein the BFC is DBCO-PEG4-NHS-Ester, DBCO-PEG4-Acid, DBCO-PEG4-Amine or DBCO-PEG4-Maleimide.
36. A pharmaceutical composition comprising the 18 F-radiolabeled protein-based probe of claim 1 , and a pharmaceutically acceptable carrier.
37. A pharmaceutical composition comprising the 18 F-radiolabeled protein-based probe of claim 12 , and a pharmaceutically acceptable carrier.
38. A pharmaceutical composition comprising the 18 F-radiolabeled protein-based probe of claim 20 , and a pharmaceutically acceptable carrier.
39. A pharmaceutical composition comprising the 18 F-radiolabeled protein-based probe of claim 24 , and a pharmaceutically acceptable carrier.
40. A pharmaceutical composition comprising the 18 F-radiolabeled protein-based probe of claim 30 , and a pharmaceutically acceptable carrier.