CROWN ETHER DERIVATIVES

• US 20070043212A1
• Filed: 10/02/2006
• Published: 02/22/2007
• Est. Priority Date: 12/20/2000
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Molecular Probes Incorporated (Thermo Fisher Scientific Incorporated)
• Inventors: GEE, Kyle, MARTIN, Vladimir
• Granted Patent: US 7,579,463 B2
• Time in Patent Office: Days
• US Class Current: 530/409
• 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 chelaters 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 selected from the group comprising of an acrylamide, an activated ester of a carboxylic acid, a carboxylic ester, an acyl azide, an acyl nitrile an aldehyde, an alkyl halide, an anhydride, an aniline, an amine, an aryl halide, an azide, an aziridine, a boronate, a diazoalkane, a haloactamide, a halotriazine, a hydrazine, an imido ester, an isocyantate, an isothiocyanate, a maleimide, a phosphoramidite, a reactive platinum complex, a silyl halide, a sulfony halide, a thiol and a photoactivatable group.

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

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

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

Sc is 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 or a virus.

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

Rx is 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, or a tyramide.

6. The compound according to claim 1, wherein said -DYE is selected from the group consisting of xanthene, borapolyazaindacene, carbocyanine, benzofuran, quinazolinone, indole, a benzazole, oxazine, and coumarin.

7. The compound according to claim 6, wherein said -DYE moiety is independently substituted by a lipophilic group.

8. The compound according to claim 7, wherein said lipophillo group is an AM or acetate ester.

9. The compound according to claim 1, wherein said DYE is a xanthene,

10. The compound according to claim 9, wherein said DYE moiety is independently substituted by a lipophilic group.

11. The compound according to claim 10, wherein said lipophilic group is an AM or acetate ester.

12. The compound according to claim 1, wherein R⁷, R⁸, R⁹, R¹⁰, R¹⁹ and R²⁰, when present, are H.

13. 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.

14. The compound according to claim 13, wherein said R¹ is methyl or ethyl.

15. The compound according to claim 13 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.

16. The compound according to claim 15 wherein said R¹⁶ is methyl;

17. The compound according to claim 13 wherein said R¹⁷ and R¹⁸ are each methyl.

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

19. A composition comprising:

a. a compound according to claim 1;

and,

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

20. The composition according to claim 19, wherein said metal ion is selected from the group consisting of Li⁺ and K⁺.

21. 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.

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

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

23. A kit for binding a metal on in a sample, comprising:

a compound of claim 1;

and, comprising one or more components selected from the group consisting of a calibration standard of a metal ion, an ionophore, a fluorescent standard, an aqueous buffer solution and an organic solvent.

24. A crown ether chelating compound having formula;