In vivo copper-free click chemistry for delivery of therapeutic and/or diagnostic agents
The present application discloses compositions and methods of synthesis and use involving click chemistry reactions for in vivo or in vitro formation of therapeutic and/or diagnostic complexes. Preferably, the diagnostic complex is of use for 18 F imaging, while the therapeutic complex is of use for targeted delivery of chemotherapeutic drugs or toxins. More preferably, a chelating moiety or targetable construct may be conjugated to a targeting molecule, such as an antibody or antibody fragment, using a click chemistry reaction involving cyclooctyne, nitrone or azide reactive moieties. In most preferred embodiments, the click chemistry reaction occurs in vivo. In vivo click chemistry is not limited to 18 F labeling but can be used for delivering a variety of therapeutic and/or diagnostic agents.
1. A method of PET imaging comprising:
a) attaching a metal- 18 F complex to a chelating moiety;
b) attaching the chelating moiety to a molecule by a click chemistry reaction in vitro to form an 18 F-labeled molecule;
c) administering the molecule to a subject; and
d) imaging the distribution of the 18 F-labeled molecule by PET (positron emission tomography).
2. The method of claim 1 , wherein the click chemistry reaction is selected from the group consisting of: (i) a nitrone with a cycloalkyne; and (ii) an azide with a cycloalkyne.
3. The method of claim 2 , wherein the cycloalkyne is cyclooctyne.
4. The method of claim 1 , wherein the molecule is a protein or peptide.
5. The method of claim 1 , wherein the molecule is a targeting molecule selected from the group consisting of an antibody, a monoclonal antibody, a bispecific antibody, a multispecific antibody, an antibody fusion protein, an antigen-binding antibody fragment and an affibody.
6. The method of claim 1 , wherein the molecule is a targetable construct.
7. The method of claim 6 , further comprising:
e) administering a targeting molecule to a subject;
f) allowing sufficient time for the targeting molecule to bind to a target antigen; and
g) subsequently administering the targetable construct to the subject, wherein the targetable construct binds to the targeting molecule.
8. The method of claim 1 , wherein the metal is selected from the group consisting of aluminum, gallium, indium, lutetium, and thallium.
9. The method of claim 8 , wherein the metal is aluminum.
10. The method of claim 1 , wherein the chelating moiety is selected from the group consisting of DOTA, TETA, NOTA, NODA, (tert-Butyl) 2 NODA, NETA, C-NETA, L-NETA, S-NETA, NODA-MPAA, and NODA-MPAEM.
11. The method of claim 1 , wherein an organic solvent is added to the medium to attach the metal- 18 F complex to the chelating moiety.
12. The method of claim 1 , wherein the metal- 18 F complex is attached to the chelating moiety by heating or microwave irradiation.
13. A method of PET imaging comprising:
a) attaching a metal- 18 F complex to a chelating moiety;
b) administering a targeting molecule to a subject;
c) allowing the targeting molecule to localize to a target cell, tissue, organ or pathogen; and
d) administering the chelating moiety bound to the metal- 18 F complex to the subject;
wherein the chelating moiety binds to the targeting molecule by a click chemistry reaction in vivo.
14. The method of claim 13 , wherein the click chemistry reaction is selected from the group consisting of: (i) a nitrone with a cycloalkyne; and (ii) an azide with a cycloalkyne.
15. The method of claim 14 , wherein the cycloalkyne is cyclooctyne.
16. The method of claim 13 , wherein the targeting molecule is selected from the group consisting of an antibody, a monoclonal antibody, a bispecific antibody, a multispecific antibody, an antibody fusion protein, an antigen-binding antibody fragment and an affibody.
17. The method of claim 13 , wherein the metal is selected from the group consisting of aluminum, gallium, indium, lutetium, and thallium.
18. The method of claim 17 , wherein the metal is aluminum.
19. The method of claim 13 , wherein the chelating moiety is selected from the group consisting of DOTA, TETA, NOTA, NODA, (tert-Butyl) 2 NODA, NETA, C-NETA, L-NETA, S-NETA, NODA-MPAA, and NODA-MPAEM.
20. A method of PET imaging comprising:
a) attaching a metal- 18 F complex to a chelating moiety;
b) attaching the chelating moiety to a targetable construct to form an 18 F-labeled targetable construct;
c) administering a targeting molecule to a subject, wherein the targeting molecule localizes to a target cell, tissue, organ or pathogen; and
d) administering the targetable construct to the subject, wherein the targetable construct binds to the targeting molecule by a click chemistry reaction in vivo.
21. The method of claim 20 , wherein the click chemistry reaction is selected from the group consisting of: (i) a nitron with a cycloalkyne; and (ii) an azide with a cycloalkyne.
22. The method of claim 21 , wherein the cycloalkyne is cyclooctyne.
23. The method of claim 21 , wherein the targeting molecule is selected from the group consisting of an antibody, a monoclonal antibody, a bispecific antibody, a multispecific antibody, an antibody fusion protein, an antigen-binding antibody fragment and an affibody.
24. The method of claim 21 , wherein the metal is selected from the group consisting of aluminum, gallium, indium, lutetium, and thallium.
25. The method of claim 24 , wherein the metal is aluminum.
26. The method of claim 21 , wherein the chelating moiety is selected from the group consisting of DOTA, TETA, NOTA, NODA, (tert-Butyl) 2 NODA, NETA, C-NETA, L-NETA, S-NETA, NODA-MPAA, and NODA-MPAEM.