Luminescence indicator for determining calcium ions

• US 6,171,866 B1
• Filed: 09/30/1998
• Issued: 01/09/2001
• Est. Priority Date: 09/30/1998
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics Corporation (Roche Holding AG)
• Original Assignee: AVL Medical Instruments
• Inventors: Mortellaro, Mark Alan, He, Huarui
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US09/164,516
• Time in Patent Office: 832 Days
• Field of Search: 436/73, 436/79, 436/172, 422/82-5-, 422/56, 562/443, 562/433, 562/405
• US Class Current: 436/79
• CPC Class Codes: C07C 217/84 the oxygen atom of at least one of the etherified hydroxy groups being further bound to an acyclic carbon atom
• CPC Class Codes: C07C 229/16 to carbon atoms of hydrocarbon radicals substituted by amino or carboxyl groups, e.g. ethylenediamine-tetra-acetic acid, iminodiacetic acids
• CPC Class Codes: C07C 237/12 having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups peptides C07K
• CPC Class Codes: C07C 271/28 to a carbon atom of a non-condensed six-membered aromatic ring
• CPC Class Codes: C07D 221/14 Aza-phenalenes, e.g. 1,8-naphthalimide
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence

Abstract

The invention relates to a compound having the general Formula I:

including its salts, where Z is either the group having the general Formula II:

where

R₁ is alkyl having 1-4 C atoms, alkoxyalkyl having 2-5 C atoms or aryloxyalkyl whose alkyl group has 1-4 C atoms,

R₂ is alkyl having 1-4 C atoms or alkoxyalkyl having 2-5 C atoms,

R₃ is H, alkoxy having 1-4 C atoms, halogen, NO or NO₂,

Y is H₂ or O and

L is a luminophoric moiety in a position para or meta to the nitrogen,

or is the group having the general Formula III:

where

n is 2 or 3,

R₄ is alkyl having 1-4 C atoms or alkoxyalkyl having 2-5 C atoms,

R₅ is H, alkoxy having 1-4 C atoms, halogen, NO or NO₂ and

L is a luminophoric moiety in a position para or meta to the nitrogen,

or is the group having the general Formula IV:

where

R₆ is alkyl having 1-3 C atoms or phenyl,

R₇ is H, alkoxy having 1-4 C atoms, halogen, NO or NO₂ and

L is a luminophoric moiety in a position para or meta to the nitrogen.

The compound of the invention can be used as a luminescence indicator for the determination of calcium ions in a sample.

Claims

1. Compound selected from the group consisting of a compound having the general Formula I:

and salts thereof, wherein Z is selected from the group consisting of a group having the general Formula II;

wherein

R₁ is selected from the group consisting of alkyl having 1-4 C atoms, alkoxyalkyl with 2-5 C atoms and aryloxyalkyl whose alkyl group has 1-4 C atoms,

R₂ is selected from the group consisting of alkyl having 1-4 C atoms and alkoxyalkyl having 2-5 C atoms,

R₃ is selected from the group consisting of H, alkoxy having 1-4 C atoms, halogen, NO and NO₂,

Y is selected from the group consisting of H₂ and O and

L is a luminophoric moiety in a position para or meta to the nitrogen,

a group having the general Formula III;

wherein

n is selected from the group of 2 and 3,

R₄ is selected from the group consisting of alkyl having 1-4 C atoms and alkoxyalkyl having 2-5 C atoms,

R₅ is selected from the group consisting of H, alkoxy having 1-4 C atoms, halogen, NO and NO₂ and

L is a luminophoric moiety in a position para or meta to the nitrogen,

and a group having the Formula IV;

wherein

R₆ is selected from the group consisting of alkyl having 1-3 C atoms and phenyl,

R₇ is selected from the group consisting of H, alkoxy having 1-4 C atoms, halogen, NO and NO₂ and

L is a luminophoric moiety in a position para or meta to the nitrogen.

2. Compound according to claim 1 having a group having the general Formula (II), R₁ is selected from the group consisting of alkyl having 4 C atoms, alkoxyalkyl having 3-4 C atoms and aryloxyalkyl whose alkyl group has 2 C atoms.

3. Compound according to any one of claim 1 or claim 2 having a group having, the general Formula (II), wherein R₂ is alkyl having 1-4 C atoms.

4. Compound according to claim 3 wherein R₂ is methyl.

5. Compound according to claim 4 having a group having the general Formula (II) wherein R₃ is located in a position para to the oxygen of an o-anisidine.

6. Compound according to claim 4 having a group having the general Formula (II) wherein R₃ is selected from the group consisting of H and methoxy.

7. Compound according to claim 4 having a group having the general Formula (III) wherein R₄ is alkyl having 1-2 C atoms.

8. Compound according to claim 4 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

9. Compound according to claim 4 having a group having the general Formula (III) wherein R₅ is selected from the group consisting of H, ethoxy and Cl.

10. Compound according to claim 3 having a group having the general Formula (II) wherein R₃ is located in a position para to the oxygen of an o-anisidine.

11. Compound according to claim 3 having a group having the general Fomula (II) wherein R₃ is selected from the group consisting of H and methoxy.

12. Compound according to claim 3 having a group having the general Formula (III) wherein R₄ is alkyl having 1-2 C atoms.

13. Compound according to claim 3 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

14. Compound according to claim 3 having a group having the general Formula (IlI) wherein R₅ is selected from the group consisting of H, ethoxy and Cl.

15. Compound according to any one of claim 1 or claim 2 having a group having the general Formula (II), wherein R₃ is located in a position para to the oxygen of an o-anisidine.

16. Compound according to claim 15 having a group having the general Formula (II) wherein R₃ is selected from the group consisting of H and methoxy.

17. Compound according to claim 15 having a group having the general Formula (III) wherein R₄ is alkyl having 1-2 C atoms.

18. Compound according to claim 15 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

19. Compound according to claim 18 having a group having the general Formula (III) wherein R₁₅ is selected from the group consisting of H, ethoxy and Cl.

20. Compound according to any one of claim 1 or claim 2 having a group having the general Formula (II), wherein R₃ is selected from the group consisting of H and methoxy.

21. Compound according to claim 20 having a group having the general Formula (III) wherein R₄ is alkyl having 1-2 C atoms.

22. Compound according to claim 20 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

23. Compound according to claim 20 having a group having the general Formula (III) wherein R₅ is selected from the group consisting of H, ethoxy and Cl.

24. Compound according to claim 1 having a group having the general Formula (III), wherein n=2.

25. Compound according to any one of claim 1, claim 2 or claim 24 having a group having the general Formula (III), wherein R₄ is alkyl having 1-2 C atoms.

26. Compound according to claim 25 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

27. Compound according to claim 25 having a group having the general Formula (III) wherein R₁₅ is selected from the group consisting of H, ethoxy and Cl.

28. Compound according to any one of claim 1, claim 2 or claim 24 having a group having the general Formula (III) wherein R₅ is located in a position para to the oxygen of an o-anisidine.

29. Compound according to claim 28 having a group having the general Formula (III) wherein R₅ is selected from the group consisting of H, ethoxy and Cl.

30. Compound according to any one of claim 1, claim 2 or claim 24 having a group having the general Formula (III) wherein R₅ is selected from the group consisting of H, ethoxy and Cl.

31. Compound according to claim 1 having a group having the general Formula (IV) wherein R₆ is methyl.

32. Compound according to any one of claim 1 or claim 3 having, a group having the general Formula (IV) wherein R₇ is located in a position para to the oxygen of an o-anisidine.

33. Compound according to claim 32 having a group having the general Formula (IV) wherein R₇ is H.

34. Compound according to any one of claim 1 or claim 31 having a group having the general Formula (IV) wherein R₇ is H.

35. Compound according to any one of claim 1, claim 2, claim 24 or claim 31 wherein L is located in a position para to the nitrogen.

36. Compound according to any one of claim 1, claim 2, claim 24 or claim 31 wherein the compound is present in the form of the dipotassium salt.

37. Optical sensor for determining calcium ions in a sample, which sensor has a matrix comprising a compound having a luminophoric moiety and an ionophoric moiety, wherein the compound is a compound according to any one of claim 1, claim 2, claim 24 or claim 31.

38. Method for determining calcium ions in a sample comprising:

providing a compound according to any one of claim 1, claim 2, claim 24 or claim 31 having a luminophore moiety and an ionophoric moiety contacting the sample at least indirectly with the compound having a luminophoric moiety and an ionophoric moiety, wherein the ioniophoric moiety reacts with the calcium ions present in the sample, and wherein the luminophoric moiety changes its luminescence properties, measuring luminescence and

determining the amount of calcium ions in the sample using the measured luminescence.