Crown ether derivatives

• US 7,579,463 B2
• Filed: 10/02/2006
• Issued: 08/25/2009
• Est. Priority Date: 12/20/2000
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Gee, Kyle R., Martin, Vladimir V.
• Primary Examiner(s): Kifle; Bruck
• Application Number:
  US11/538,010
• Publication Number:
  US 20070043212A1
• Time in Patent Office: 1,058 Days
• Field of Search: 540/465, 540/468
• US Class Current: 540/465
• CPC Class Codes: C07D 273/00 Heterocyclic compounds containing rings having nitrogen and oxygen atoms as the only ring hetero atoms, not provided for by groups C07D261/00 - C07D271/00
• CPC Class Codes: C07D 413/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 413/14 containing three or more hetero rings

Abstract

The invention describes crown ether chelators, including crown ethers having the formula:

Claims

1. A crown ether chelating compound having the formula:

2. The compound according to claim 1, wherein said —

R_x is a radical of a reactive group selected from the group consisting of carboxylic acid, succinimidyl ester of a carboxylic acid, hydrazide, amine and a maleimide.

3. The compound according to claim 1, wherein said —

S_c is a radical of a conjugated substance selected from the group consisting of an amino acid, a peptide, a protein, a polysaccharide, a nucleoside, a nucleotide, an oligonucleotide, a nucleic acid, a hapten, a psoralen, a drug, a hormone, a lipid, a lipid assembly, a synthetic polymer, a polymeric microparticle, a biological cell and a virus.

4. The compound according to claim 1, wherein said —

S_c is a radical of a conjugated substance selected from the group consisting of an antibody or fragment thereof, an avidin or streptavidin, a biotin, a blood component protein, a dextran, an enzyme, an enzyme inhibitor, a hormone, an IgG binding protein, a fluorescent protein, a growth factor, a lectin, a lipopolysaccharide, a microorganism, a metal binding protein, a metal chelating moiety, a non-biological microparticle, a peptide toxin, a phosphotidylserine-binding protein, a structural protein, a small-molecule drug, and a tyramide.

5. The compound according to claim 1, wherein said -DYE moiety is independently substituted by a lipophilic group.

6. The compound according to claim 5, wherein said lipophilic group is an AM or acetate ester.

7. The compound according to claim 1, wherein said DYE is a radical of xanthene.

8. The compound according to claim 7, wherein said DYE moiety is independently substituted by a lipophilic group.

9. The compound according to claim 8, wherein said lipophilic group is an AM or acetate ester.

10. The compound according to claim 1, wherein R⁷, R⁸, R¹⁰, R¹⁹ and R²⁰, when present, are H.

11. The compound according to claim 1, wherein R¹ is C₁-C₆ alkyl that is substituted one or more times by amino (—

NR¹⁷R¹⁸), —

(C═

O)—

O—

R¹⁶ or —

(C═

O)—

NR¹⁷R¹⁸.

12. The compound according to claim 11, wherein said R¹ is methyl or ethyl.

13. The compound according to claim 11 wherein said R¹⁶ is selected from the group consisting of H, C₁-C₆ alkyl, benzyl, a biologically compatible esterifying group, and a biologically compatible salt.

14. The compound according to claim 13 wherein said R¹⁶ is methyl.

15. The compound according to claim 11 wherein said R¹⁷ and R¹⁸ are each methyl.

16. The compound according to claim 1, wherein said R¹⁹ and R²⁰ are H.

17. A composition comprising:

a. a compound according to claim 1; and

,

b. a metal ion that is capable of being chelated by said compound.

18. The composition according to claim 17, wherein said metal ion is selected from the group consisting of Li⁺ and K⁺.

19. A method for binding a target metal ion in a sample, comprising steps of:

a. contacting said sample with a metal chelating compound of claim 1;

b. incubating said sample and said metal chelating compound for sufficient time to allow said compound to chelate said target metal ion whereby said metal ion is bound.

20. A method for binding and detecting target ions in a live cell, said method comprising:

a) contacting a sample of live cells with a crown ether compound of claim 1 with the proviso that said compound comprise a DYE moiety and at least one lipohilic group;

b) incubating said sample and said crown ether chelate compound for sufficient time to allow said compound to chelate said target metal ion; and

,

c) illuminating said sample with an appropriate wavelength whereby said target ion is detected in a live cell.