Targeted contrast agents and methods for targeting contrast agents
View Patent ↗A contrast agent having a contrast protein have contrast properties and at least one targeting moiety, wherein the at least one targeting moiety is operatively linked to or incorporated within the contrast protein. Methods for targeting contrast agents and for preparing such agents are included.
1. A recombinant protein comprising:
domain 1 of a CD2 protein, where the domain 1 is modified to contain a paramagnetic metal binding site, the paramagnetic metal binding site comprising:
at least three mutations in domain 1 of the CD2 protein, where the at least three mutations are N15E, L58D, and K64D; and
a targeting moiety, where the targeting moiety is operatively linked to the domain 1 of the CD2 protein,
wherein the recombinant protein has contrast properties.
2. The recombinant protein of claim 1 , further comprising a linker, wherein the linker is operatively coupled to the domain 1 of the CD2 protein and the targeting moiety.
3. The recombinant protein of claim 1 , wherein the paramagnetic metal binding site further comprises a fourth mutation, where the fourth mutation is E29G.
4. The recombinant protein of claim 1 , wherein the paramagnetic metal binding site further comprises a fourth mutation, where the fourth mutation is N17D.
5. The recombinant protein of claim 1 , wherein the paramagnetic metal binding site further comprises a fourth and a fifth mutation, where the fourth mutation is N17D and where the fifth mutation is E29G.
6. The recombinant protein of claim 1 , wherein the targeting moiety is operatively coupled to the c-terminus of the recombinant protein.
7. The recombinant protein of claim 1 , wherein the targeting moiety is operatively coupled to the n-terminus of the recombinant protein.
8. The recombinant protein of claim 1 , wherein the targeting moiety is inserted between the c- and n-terminus of the recombinant protein.
9. The recombinant protein of claim 1 , wherein the targeting moiety induces endocytosis in a target cell.
10. The recombinant protein of claim 1 , wherein the targeting moiety is operatively linked to the recombinant protein by a covalent or peptide bond.
11. The recombinant protein of claim 1 , wherein the paramagnetic ion is Gd(III).
12. The recombinant protein of claim 1 , wherein the targeting moiety is selected from the group consisting of: gastrin release peptide, affibody, VIP, galinin, secretin, CCK8, angiotensin I, human endothelin 3, human neuropeptide, human opioid peptide a-endorphin, and GH-RH.
13. The method of claim 1 , wherein the recombinant protein has a sequence identical to any one of SEQ ID NOs: 1-11 and 14-26.
14. A method for preparing a contrast agent comprising:
selecting a CD2 protein;
modifying a CD2 protein to contain a paramagnetic binding site such that the domain 1 of the CD2 protein contains a N15E, a L58D, and a K64D mutation to generate a modified CD2 protein;
selecting a targeting moiety configured to bind a target protein; and
operatively linking the targeting moiety and the CD2 protein.
15. The method of claim 14 , further comprising the step of chelating a paramagnetic ion to the contrast protein by binding the paramagnetic ion directly at least to the N15E, L58D, and K64D amino acids of the modified CD2 protein.
16. The method of claim 15 , wherein the targeting moiety is selected from the group consisting of: gastrin release peptide, affibody, VIP, galinin, secretin, CCK8,angiotensin I, human endothelin 3, human neuropeptide, human opioid peptide a-endorphin, and GH-RH.
17. The method of claim 14 , wherein the targeting moiety is configured to bind a cancer cell.
18. The method of claim 14 , wherein the targeting moiety is configured to induce endocytosis in a target cell.
19. The method of claim 14 , wherein the recombinant protein has a sequence identical to any one of SEQ ID NOs.: 1-11 and 14-26.
20. A method of magnetic resonance imaging of a subject comprising:
administering a recombinant protein of claim 1 to a subject in need thereof;
allowing the recombinant protein to bind a target cell; and
imaging the subject using magnetic resonance imaging.
21. The method of claim 20 , wherein the subject is imaged a site of the target cell.
22. The method of claim 20 , wherein the recombinant protein has a sequence according to any one of SEQ ID NOs: 1-11 and 14-26.