Heat shock protein as a targeting agent for endothelium-specific in vivo transduction

• US 20060034761A1
• Filed: 08/13/2004
• Published: 02/16/2006
• Est. Priority Date: 08/13/2004
• Status: Active Grant

• Current Assignee: General Electric Company
• Original Assignee: General Electric Company
• Inventors: Zhao, Ming, Syud, Faisal A., Torres, Andrew S.
• Granted Patent: US 7,575,738 B2
• Time in Patent Office: Days
• US Class Current: 424/9.100
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07B 59/008 Peptides; Proteins

Abstract

The present invention relates to the use of Heat Shock Proteins and fragments thereof as targeting ligand. The Heat Shock Protein may be labeled with imaging agents that are capable of binding lectin-like oxidized low-density lipoprotein (LOX-1) or may be attached to a therapeutic agent. The sequences are useful for the diagnosis and monitoring of diseases as well as means for internalizing signaling moieties and therapeutics.

Claims

1. A compound having the formula S-(L)_n-H, wherein S is a signal providing structural unit that provides a signal that can be detected in vivo or detected in vitro, L links S to H, H is HSP, a fragment, or a variant thereof that binds to LOX-1, and n is either 0 or 1.

5. A method of imaging a tissue to detect the presence and/or amount of LOX-1, comprising:

administering to a mammal the compound of claim 1;

optionally administering a clearing agent to remove the compound that is not bound to LOX-1; and

subjecting the mammal to imaging effective to detect the signal generated by S to thereby detect the presence and/or amount of LOX-1.

6. A method of monitoring the efficacy of therapies for treating atherosclerosis comprising:

administering to a mammal the compound of claim 1;

optionally administering a clearing agent to remove the compound that is not bound to LOX-1;

subjecting the mammal to imaging effective to detect the signal generated by S to thereby detect the amount of LOX-1; and

repeating the administration and imaging procedures at least once over a period of time to detect the difference in amount of LOX-1.

7. The compound of claim 1, wherein S is selected from the group consisting of a luminescent dye, a radionuclide, a near infrared dye, a magnetically active isotope, a superparamagnetic particle, a metal ion having a Z value of greater than 50, an encapsulated species, and a combination thereof.

8. The compound of claim 1, wherein S is selected from the group consisting of fluorescein, ¹¹C, ¹⁸F, ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁸⁹Zr, ^94mTc, ⁹⁴Tc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ^154-158Gd, ¹⁷⁵Lu, superparamagnetic iron oxide nanoparticles, heavy metal ions, gas-filled microbubbles, optical dyes, porphyrins, texaphyrins, highly iodinated organic compounds chelates thereof, polymers containing at least one of the aforementioned components, endohedral fullerenes containing at least one of the aforementioned, and mixtures thereof.

9. The compound of claim 7, wherein S is a luminescent dye.

10. The compound of claim 9, wherein the luminescent dye is fluorescein, or derivatives thereof.

11. The compound of claim 6, wherein S is a radionuclide.

12. The compound of claim 11, wherein the radionuclide is a positron emitter.

13. The compound of claim 12, wherein the positron emitter is selected from ¹⁸F and ¹¹C.

14. The compound of claim 11, wherein the radionuclide is a gamma emitter.

15. The compound of claim 7, wherein S is an infrared dye.

16. The compound of claim 7, wherein S is a magnetically active isotope.

17. The compound of claim 16, wherein the magnetically active isotope is paramagnetic.

18. The compound of claim 17, wherein the magnetically active isotope is an isotope of gadolinium.

19. The compound of claim 7, wherein S is a superparamagnetic particle.

20. The compound of claim 19, wherein the superparamagnetic particle is a nanoparticle.

21. The compound according to claim 20, wherein the nanoparticle comprises at least one of iron oxide and elemental iron.

22. The compound according to claim 7, wherein S is an element having a Z value of greater than about 50.

23. The compound according to claim 22, wherein the element having a Z value of greater than about 50 is iodine or bismuth.

24. The compound according to claim 7, wherein S is an encapsulated species.

25. The compound according to claim 24, wherein the encapsulated species is selected from the group consisting of a micelle, a liposome, a polysome, and a gas-filled microbubble.

26. The compound according to claim 1, wherein L is an organic radical having a valence of at least 2.

27. The compound according to claim 26, wherein the organic radical is covalently bound to both group S and group H.

28. The compound according to claim 26, wherein the organic radical is ionically bound to one of group S and group H.

29. The compound according to claim 28, wherein the organic radical is ionically bound to both group S and group H.

30. The compound according to claim 26, wherein the organic radical comprises between 1 and about 10,000 carbon atoms.

31. The compound according to claim 30, wherein the organic radical is selected from the group consisting of alkylene, arylene, cycloakylene, aminoaklylene, aminoarylene, aminocycloalkylene, thioalkylene, thioarylene, thiocycloalkylene, oxyalkylene, oxyarylene, oxycycloalkylene, acylalkylene, acylarylene, acylcycloalkylene units, and combinations thereof.

32. The compound of claim 31, wherein the acylarylene unit is a 4-acylphenylene group having the following structure:

33. The compound of claim 26, wherein the organic radical is a metal chelating agent.

34. The compound according to claim 33, wherein the metal chelating agent binds at least one metal cation selected from the group consisting of cations of ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, 86Y, ⁸⁹Zr, ^94mTc, ⁹⁴Tc, ^99mTc, ¹¹¹In, 154-158Gd, and ¹⁷⁵Lu.

35. The compound of claim 33, wherein the metal chelating agent is selected from the group consisting of DTPA, 1,4,7-triaza-cyclononane-N,N′

,N″

-triacetic acid (NOTA), p-bromoacetamido-benyl-tetraethylaminetetraacetic acid (TETA), 1,4,7,10-tetraazacyclododecanetetraacetic acid (DOTA), EDTA, CHXa.

36. The compound according to claim 1, wherein H is Heat Shock Protein 70.

37. The compound according to claim 1, wherein H is a fragment of Heat Shock Protein 70.

38. The compound according to claim 1, wherein H is an N-terminus fragment of Heat Shock Protein 70.

43. The method of claim 5, wherein the mammal is suspected of a disease or disorder caused by expression of LOX-1.

44. The method of claim 5, wherein the imaging effective to detect S is positron emission tomography.

45. The method of claim 44, wherein S is selected from ¹⁸F and ¹¹C.

46. The method of claim 6, wherein the mammal is suspected of a disease or disorder caused by expression of LOX-1.

47. The method of claim 6, wherein the imaging effective to detect S is positron emission tomography.

48. The method of claim 6, wherein S is selected from ¹⁸F and ¹¹C.

2. A compound having the formula T-(L)_n-H, wherein T is a therapeutic agent, L links T to H, H is HSP or a fragment thereof that binds to LOX-1, and n is either 0 or 1.

3. A composition comprising a compound having the formula S-(L)_n-H, wherein S is a signal providing structural unit that provides a signal that can be detected in vivo or detected in vitro, L links S to H, H is HSP, a fragment, or a variant that binds to LOX-1, and n is either 0 or 1.

39. The composition of claim 3, wherein S is selected from the group consisting of a luminescent dye, a radionuclide, a near infrared dye, a magnetically active isotope, a superparamagnetic particle, a metal ion having a Z value of greater than 50, an encapsulated species, and a combination thereof.

40. The composition of claim 3, wherein S is selected from the group consisting of fluorescein, ¹¹C, ¹⁸F, ⁵²Fe, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁸⁹Zr, ⁹⁴Tc, ⁹⁴Tc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ^154-158Gd, ¹⁷⁵Lu, superparamagnetic iron oxide nanoparticles, heavy metal ions, gas-filled microbubbles, optical dyes, porphyrins, texaphyrins, highly iodinated organic compounds chelates thereof, polymers containing at least one of the aforementioned components, endohedral fullerenes containing at least one of the aforementioned, and mixtures thereof.

41. The composition of claim 40, wherein S is selected from ¹⁸F and ¹¹C.

42. A kit comprising the composition of claim 3.

4. A composition comprising a compound having the formula T-(L)_n-H, wherein T is a therapeutic agent, L links T to H, H is HSP, a fragment, or a variant that binds to LOX-1, and n is either 0 or 1.