Luminescent macrocyclic lanthanide complexes

• US 8,173,800 B2
• Filed: 08/15/2007
• Issued: 05/08/2012
• Est. Priority Date: 08/15/2006
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Raymond, Kenneth N., Corneillie, Todd M., Xu, Jide
• Primary Examiner(s): Ward, Paul V.
• Application Number:
  US11/839,509
• Publication Number:
  US 20080213917A1
• Time in Patent Office: 1,728 Days
• Field of Search: 540/460, 540/140, 530/391.3, 435/24, 564/152, 436/546
• US Class Current: 540/145
• CPC Class Codes: G01N 2458/40 Rare earth chelates
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The present invention provides a novel class of macrocyclic compounds as well as complexes formed between a metal (e.g., lanthanide) ion and the compounds of the invention. Preferred complexes exhibit high stability as well as high quantum yields of lanthanide ion luminescence in aqueous media without the need for secondary activating agents. Preferred compounds incorporate hydroxy-isophthalamide moieties within their macrocyclic structure and are characterized by surprisingly low, non-specific binding to a variety of polypeptides such as antibodies and proteins as well as high kinetic stability. These characteristics distinguish them from known, open-structured ligands.

Claims

1. A compound having a structure according to Formula I:

2. The compound according to claim 1, having the structure:

3. The compound according to claim 1, wherein at least one of L¹, L², L³, L⁴, L⁵, L⁶, L⁷, L⁸, L⁹, L¹⁰, A¹, A², A³ and A⁴ is substituted with a functional moiety.

4. The compound according to claim 1, wherein said functional moiety has the structure:

5. The compound according to claim 4, having a structure, which is a member selected from:

6. The compound according to claim 4, wherein said targeting moiety comprises a member selected from a small-molecule ligand, a peptide, a protein, an enzyme, an antibody, an antigen, a nucleic acid, a carbohydrate, a lipid and a pharmacologically active molecule.

7. The compound according to claim 4, wherein said targeting moiety is substituted with a luminescence modifying group allowing luminescence energy transfer between said complex and said luminescence modifying group when said complex is excited.

8. The compound according to claim 4, wherein said reactive functional group is a member selected from —

OH, —

SH, —

NH₂, —

C(O)NHNH₂ (hydrazide), maleimide, activated ester, aldehyde, ketone, hydroxylamine, imidoester, isocyanate, isothiocyanate, sulfonylchloride, acylhalide and —

COOY, wherein Y is a member selected from H, a negative charge and a salt counter-ion.

9. The compound according to claim 4, wherein X is NH₂ and L¹¹ is a member selected from substituted or unsubstituted C₁ to C₁₀ alkyl and substituted or unsubstituted C₁ to C₁₀ heteroalkyl.

10. The compound according to claim 4, wherein said functional moiety comprises a polyether.

11. The compound according to claim 10, wherein said polyether is a member selected from polyethylene glycol (PEG) and derivatives thereof.

12. The compound according to claim 11, wherein said polyether has a molecular weight of about 50 to about 10,000 daltons.

13. The compound according to claim 1, wherein said linker moieties L¹, L², L³, L⁴, L⁵, L⁶, L⁷, L⁸, L⁹ and L¹⁰ are members independently selected from substituted or unsubstituted C₁ to C₆ alkyl.

14. The compound according to claim 13, wherein said linker moieties L¹, L², L³, L⁴, L⁵, L⁶, L⁷, L⁸, L⁹ and L¹⁰ are members independently selected from substituted or unsubstituted ethyl.

15. A luminescent complex formed between at least one metal ion and a compound according to claim 1.

16. The complex according to claim 15, wherein said metal ion is a lanthanide ion.

17. The complex according to claim 16, wherein said lanthanide is a member selected from neodynium (Nd), samarium (Sm), europium (Eu), terbium (Tb), dysprosium (Dy) and ytterbium (Yb).

18. A method of detecting the presence of an analyte in a sample, said method comprising:

(a) contacting said sample and a composition comprising a luminescent complex according to claim 15,

(b) exciting said complex; and

(c) detecting luminescence from said complex.

19. A method of detecting the presence of an analyte in a sample, said method comprising:

(a) contacting said sample and a composition comprising a luminescent complex according to claim 15 and a luminescence modifying group, wherein energy can be transferred between said luminescent complex and said luminescence modifying group when said complex is excited, and wherein said complex and said luminescence modifying group can be part of the same molecule or be part of different molecules;

(b) exciting said complex; and

(c) determining the luminescent property of said sample, wherein the presence of said analyte results in a change in said luminescent property.

20. The method according to claim 19, wherein said analyte, if present in said sample, binds to an antibody, wherein said antibody is covalently linked to a member selected from a luminescence modifying group and a complex according to claim 15.

21. The method according to claim 20, wherein said analyte is a lipid.