Methods and compositions for imaging
View Patent ↗The present invention is directed to compositions useful as imaging agents for use in monitoring atherosclerotic plaque regression using, for example, MRI, CT, Gamma-scintigraphy, or optical imaging techniques. Methods and compositions of using the same are described.
1. A synthetic nanoparticle, said synthetic nanoparticle comprising: at least one apolipoprotein that is not an apoB lipoprotein, at least one amphipathic lipid, and at least one metallic or non-metallic contrast agent conjugated to a component of said nanoparticle, said at least one metallic or non-metallic contrast agent being present in an amount between 5% to about 50% (w/w) of said nanoparticle, and said synthetic nanoparticle having a diameter from about 5 nm to about 50 nm,
and wherein said nanoparticle further comprises a C—C chemokine receptor type 7 (CCR7) targeting moiety for monitoring atherosclerosis.
2. The synthetic nanoparticle of claim 1 , wherein said nanoparticle is discoidal.
3. The synthetic nanoparticle of claim 1 , wherein said nanoparticle is spherical.
4. The synthetic nanoparticle of claim 1 , wherein said apolipoprotein is an amphipathic apolipoprotein or a fragment thereof.
5. The synthetic nanoparticle of claim 1 , wherein said metallic contrast agent is selected from the group consisting of Gd(III), Mn(II), Mn(III), Cr(II), Cr(III), Cu(II), Fe (III), Pr(III), Nd(III) Sm(III), Tb(III), Yb(III) Dy(III), Ho(III), Eu(II), Eu(III), Er(III), Indium (In), Technetium (Tc), and Barium.
6. The synthetic nanoparticle of claim 1 , wherein said non-metallic contrast agent is selected from the group consisting of Iodine (I), Bromine, Fluorescein, Carboxyfluorescein, and Calcein.
7. The synthetic nanoparticle of claim 1 , wherein said metallic contrast agent is gadolinium.
8. The synthetic nanoparticle of claim 1 , wherein said metallic contrast agent is conjugated to a lipid component of the synthetic nanoparticle.
9. The synthetic nanoparticle of claim 1 , wherein said non-metallic contrast agent is conjugated to a lipid component of the synthetic nanoparticle.
10. The synthetic nanoparticle of claim 8 , wherein said lipid component of the synthetic nanoparticle is selected from the group consisting of a sterol, a phospholipid, a sterol ester, a diacylglycerol and a triacylglycerol.
11. The synthetic nanoparticle of claim 10 , wherein said phospholipid is selected from the group consisting of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), cardiolipin (CL), a sphingolipid, sphingomyelin (SM), and phosphatidic acid (PA).
12. The synthetic nanoparticle of claim 11 , wherein said phospholipid is PC.
13. The synthetic nanoparticle of claim 12 , wherein said PC is 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC).
14. The synthetic nanoparticle of claim 11 , wherein said phospholipid is PE.
15. The synthetic nanoparticle of claim 14 , wherein said PE is poly-lysine PE.
16. The synthetic nanoparticle of claim 15 , wherein said poly-lysine PE is poly-lysine dimyristoyl-PE (DMPE).
17. The synthetic nanoparticle of claim 10 , wherein said sterol is cholesterol.
18. The synthetic nanoparticle of claim 10 , wherein said sterol ester is cholesteryl ester.
19. The synthetic nanoparticle of claim 1 , wherein said metallic contrast agent is conjugated to a protein component of the synthetic nanoparticle.
20. The synthetic nanoparticle of claim 1 , wherein said non-metallic contrast agent is conjugated to a protein component of the synthetic nanoparticle.
21. The synthetic nanoparticle of claim 19 or claim 20 , wherein said protein is selected from the group consisting of an apolipoprotein A-I, A-II, A-IV, C-I, C-II, C-III and E.
22. The synthetic nanoparticle of claim 1 , comprising a phospholipids:steryl ester:sterol:TAG:ApoAI ratio (w/w) of 100:62:25:11:2.
23. The synthetic nanoparticle of claim 1 , wherein said imaging agent comprises between 1 and 50 metallic contrast agent molecules per synthetic nanoparticle.
24. The synthetic nanoparticle of claim 1 or claim 23 , wherein said metallic contrast agent molecule is conjugated to a phospholipid moiety and said phospholipid moiety accommodates more than one metallic agent molecule.
25. The synthetic nanoparticle of claim 1 or claim 23 , wherein said non-metallic contrast agent molecule is conjugated to a phospholipid moiety and said phospholipid moiety accommodates more than one metallic agent molecule.
26. The synthetic nanoparticle of claim 1 , wherein said synthetic nanoparticle comprises 10 metallic contrast agent molecules per synthetic nanoparticle.
27. The synthetic nanoparticle of claim 1 , wherein said synthetic nanoparticle comprises between about 5 mg and about 30 mg of apolipoprotein.
28. The synthetic nanoparticle of claim 1 , wherein said synthetic nanoparticle comprises between about 80 and about 90 phospholipids per synthetic nanoparticle.
29. The synthetic nanoparticle of claim 1 , wherein said synthetic nanoparticle comprises about 2 apolipoprotein molecules per synthetic nanoparticle.
30. The synthetic nanoparticle of claim 1 , wherein said synthetic nanoparticle comprises about 1 apolipoprotein molecule to about 40 phospholipid molecules.
31. The synthetic nanoparticle of claim 1 , wherein said CCR7 targeting moiety is an antibody directed against CCR7.
32. The synthetic nanoparticle of claim 1 , wherein said CCR7 targeting moiety is conjugated to a detectable label.
33. The synthetic nanoparticle of claim 1 , wherein said diameter is between about 5 to about 18 nm.
34. The synthetic nanoparticle of claim 1 , wherein said diameter is between about 5 to about 12 nm.
35. The synthetic nanoparticle of claim 1 , wherein said diameter is less than 10 nm.
36. The synthetic nanoparticle of claim 1 , comprising two or more different metallic contrast agents.
37. The synthetic nanoparticle of claim 1 , comprising two or more different non-metallic contrast agents.
38. The synthetic nanoparticle of claim 1 , further comprising a drug.
39. The synthetic nanoparticle of claim 1 , wherein said nanoparticle is non-inflammatory.
40. A composition comprising the synthetic nanoparticle of claim 1 and a pharmaceutically acceptable carrier or diluent.
41. A method of in vivo imaging of a site of atherosclerosis within a subject comprising administering to said subject a synthetic nanoparticle comprising a metallic or non-metallic contrast agent conjugated to a component of a synthetic nanoparticle according to claim 1 .
42. A method of determining atherosclerotic plaque regression comprising administering a synthetic nanoparticle according to claim 1 and determining if expression or activity of CCR7 in an atherosclerotic plaque is increased compared to a control.
43. The method of claim 42 , wherein said determining comprises determining an increase in CCR7 expression or activity using an anti-CCR7 antibody, an aptmer against CCR7 protein or mRNA, or a CCR7 ligand.
44. A method of monitoring the efficacy of an atherosclerotic drug therapy comprising contacting a subject receiving said therapy with a synthetic nanoparticle according to claim 1 and determining the regression of atherosclerotic plaques in said subject wherein an increase in CCR7 expression or activity indicates that said therapy is effective at promoting atherosclerotic plaque regression in said subject.
45. The synthetic nanoparticle of claim 1 , wherein the contrast agent is conjugated to a component of said synthetic nanoparticle core.
46. The synthetic nanoparticle of claim 45 , wherein the contrast agent is a metallic crystal or particulate.
47. The synthetic nanoparticle of claim 46 , wherein the contrast agent is a metal oxide or a quantum dot.
48. The synthetic nanoparticle of claim 45 , further comprising a second metallic or non-metallic contrast agent conjugated to a component of said nanoparticle.
49. The synthetic nanoparticle of claim 48 , wherein said first and second contrast agents are different agents.
50. A composition comprising the synthetic nanoparticle of claim 45 and a pharmaceutically acceptable carrier or diluent.
51. A method of in vivo imaging of a site of atherosclerosis within a subject comprising administering to said subject the composition of claim 50 and imaging the site of said subject.
52. The synthetic nanoparticle of any one of claims 1 , 8 , 9 , 19 , 20 , 24 , 25 , 45 and 48 , wherein said contrast agent is conjugated through a chelator to a component of said nanoparticle.