Macrocyclic HOPO chelators

• US 10,352,938 B2
• Filed: 02/25/2016
• Issued: 07/16/2019
• Est. Priority Date: 08/24/2009
• Status: Active Grant

• Current Assignee: Lumiphore Inc.
• Original Assignee: Lumiphore Inc.
• Inventors: Butlin, Nathaniel G., Raymond, Kenneth N., Xu, Jide, D'Aleo, Anthony
• Primary Examiner(s): Xu, Xiaoyun R
• Application Number:
  US15/053,713
• Publication Number:
  US 20160320399A1
• Time in Patent Office: 1,237 Days
• Field of Search: None
• CPC Class Codes: C07D 487/18 Bridged systems
• CPC Class Codes: C07D 498/22 in which the condensed system contains four or more hetero rings
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/533 with fluorescent label
• 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 comprise hydroxypyridinonyl 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.

Claims

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

2. A complex formed between at least one metal ion and the compound of claim 1.

3. A method of detecting an analyte in a sample, said method comprising:

(a) contacting said analyte with a solid support comprising the complex of claim 2, wherein said analyte forms an analyte complex;

(b) exciting said complex such that said complex transfers excitation energy to said analyte complex; and

(c) detecting energy emitted by said analyte complex, thereby detecting said analyte.

4. The compound of claim 1, wherein A¹, A², A³ and A⁴ are independently selected from:

5. The compound of claim 1, wherein A¹, A², A³ and A⁴ are each independently selected from:

6. The compound of claim 1, wherein A¹, A², A³ and A⁴ have the same relative orientation;

and

(a) A¹, A², A³ and A⁴ are each independently

7. The compound of claim 1, wherein Z is O.

8. The compound of claim 1, wherein R⁵ is unsubstituted alkyl.

9. The compound of claim 1, wherein R⁵ is methyl.

10. The compound of claim 1, wherein R⁵, R⁶ or both are H.

11. The compound of claim 1, wherein R¹ is H or a negative charge.

12. The compound of claim 1, wherein L³ is substituted or unsubstituted alkyl.

13. The compound of claim 1, wherein L³ is substituted or unsubstituted heteroalkyl.

14. The compound of claim 1, wherein

L³ comprises —

(CH₂CH₂O)_mR³¹—

;

wherein

m is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8 and 9; and

R³¹ is selected from substituted or unsubstituted alkyl and substituted or unsubstituted heteroalkyl.

15. The compound of claim 1, wherein L¹, L², L⁴, and L⁵ are independently selected substituted or unsubstituted C₁to C₆alkyl.

16. The compound of claim 1, wherein L¹, L², L⁴, and L⁵ are independently selected substituted or unsubstituted ethyl.

17. The compound of claim 1, wherein one or more groups selected from L¹, L², L³, L⁴ and L⁵ is ethyl substituted by -L¹¹-X and each of the unselected groups is unsubstituted ethyl.

18. The compound of claim 1, wherein L³ is substituted by -L¹¹X.

19. The compound of claim 1, wherein L³ is selected from —

(CH₂CH₂O)₃(CH₂)₂—

, —

(CH₂CH₂O)₄(CH₂)₂—

and —

(CH₂CH₂O)₅(CH₂)₂—

, wherein any H is optionally replaced by -L¹¹-X.

20. The compound of claim 1, wherein L¹¹ is selected from substituted or unsubstituted alkyl and substituted or unsubstituted heteroalkyl.

21. The compound of claim 1, wherein X is selected from —

NH₂and —

C(O)OH.

22. The compound of claim 1, wherein X is a linkage fragment bound to a species selected from a fluorophore, a carrier moiety and a solid support.

23. The compound of claim 22, wherein said carrier moiety is selected from a nucleic acid, a peptide, an antibody, an antibody fragment, an antigen, a receptor, a lectin, a saccharide, and a lipid.

24. The compound of claim 1, wherein -L¹¹-X is selected from:

25. The compound of claim 1, wherein R¹⁹, R²⁰, R²¹and R²²are H.

26. The compound of claim 6, wherein A¹, A², A³ and A⁴ are each independently:

27. The complex of claim 2, wherein said metal ion is a lanthanide ion.

28. The complex of claim 27, wherein said lanthanide is a member selected from neodynium (Nd), samarium (Sm), europium (Eu), gadolinium (Gd), terbium (Tb), dysprosium (Dy) and ytterbium (Yb).