Mixed ligand complexes and uses thereof as binding agents to DNA

• US 5,439,794 A
• Filed: 02/28/1992
• Issued: 08/08/1995
• Est. Priority Date: 01/18/1985
• Status: Expired due to Term

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Barton, Jacqueline K.
• Primary Examiner(s): Parr, Margaret
• Assistant Examiner(s): Marschel, Ardin H.
• Application Number:
  US07/843,315
• Time in Patent Office: 1,257 Days
• Field of Search: 435/6, 435/91, 435/968, 435/52, 435/91.1, 435/91.2, 435/810, 436/56, 436/800, 436/501, 536/23.1, 536/22.1, 536/24.1, 536/24.3-24.33, 536/25.3, 935/78, 935/88, 204/157.71, 546/88, 546/10
• US Class Current: 435/6.11
• CPC Class Codes: C07D 471/04 Ortho-condensed systems
• CPC Class Codes: C07F 15/0013 without a metal-carbon linkage
• CPC Class Codes: C07F 15/0053 without a metal-carbon linkage
• CPC Class Codes: C07F 15/008 without a metal-carbon linkage
• CPC Class Codes: C07F 15/065 without a metal-carbon linkage
• CPC Class Codes: C07F 3/003 without C-Metal linkages
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence

Abstract

This invention concerns a coordination complex or salt thereof which is spectroscopically or photoactively determinable when bound to DNA having the formula ##STR1## wherein M is a suitable transition metal and each of R₁, R₂ and R₃ is ethylenediamine or a substituted derivative thereof, bipyridine or a substituted derivative thereof, phenanthroline or a substituted derivative thereof, diazfluorene-9-one or a substituted derivative thereof, phenanthrenequinonediimine or a substituted derivative thereof; wherein R₁, R₂ and R₃ are bound to M by coordination bonds wherein R₁ and R₂ are the same and both are different from R₃. The invention also concerns a method of labeling DNA with the coordination complex, a DNA molecule labeled with the coordination complex, a method of selectively labeling DNA conformation with the coordination complex and a method of detecting the presence of a conformation present in a double stranded DNA.

Claims

1. A method for labeling DNA with a complex which is determinable spectroscopically or photoactively when bound to the DNA which comprises contacting the DNA with a coordination complex or salt thereof which is spectroscopically or photoactively determinable when bound to DNA having the formula ##STR5## wherein M is Ru, Rh, Co, Fe, Cr, Cu, Zn, Cd, or Pb and each of R₁, R₂ and R₃ is ethylenediamine or a substituted derivative thereof, bipyridine or a substituted derivative thereof, phenanthroline or a substituted derivative thereof, diazafluorene-9-one or a substituted derivative thereof, phenanthrenequinonediimine or a substituted derivative thereof;

wherein R₁, R₂ and R₃ are bound to M by coordination bonds and wherein R₁ and R₂ are the same and both are different from R₃ ;

under conditions such that the complex binds to the DNA and thereby labels the DNA with the complex.

2. The method of claim 1, wherein bipyridine is 2,2'"'"'-bipyridine (bpy), phenanthroline is 1,10-phenanthroline (phen), diazafluorene-9-one is 4,5-diazafluorene-9-one (flone), phenanthrenequinonediimine is 9,10-phenanthrenequinonediimine (phi).

3. The method of claim 1, wherein the substituted derivatives of phenanthroline comprises 4,7-diamino-1,10-phenanthroline, 3,8-diamino-1,10-phenanthroline, 4,7-diethylenediamine-1,10-phenanthroline, 3,8-diethylenediamine-1,10-phenanthroline, 4,7-dihydroxyl-1,10-phenanthroline, 3,8-dihydroxyl-1,10-phenanthroline, 4,7-dinitro-1,10-phenanthroline, 3,8-dinitro-1,10-phenanthroline, 4,7-diphenyl-1,10-phenanthroline (DIP), 3,8-diphenyl-1,10-phenanthroline, 4,7-dispermine-1,10-phenanthroline, 3,8-dispermine-1,10-phenanthroline, 5-nitro-phenanthroline (5-NO₂ phen), or 3,4,7,8-tetramethylphenanthroline (TMP) and wherein the substituted derivative of bipyridine comprises 4,4'"'"' diphenyl bipyridine, bis 4,4'"'"' methyl bipyridylate, or bis 4,4'"'"' bipyridylamide.

4. The method of claim 3, wherein the complex has the formula M(phen)₂ (phi), M(bpy)₂ (phi), M(phi)₂ (bpy), M(phi)₂ (4,4'"'"'diphenyl bipyridine), M(bis 4,4'"'"' methyl bipyridylate)₂ (phi), M(bis 4,4'"'"' bipyridylamide)₂ (phi), M(bpy)₂ (phen), M(phen)₂ (bpy), M(phen)₂ (flone), M(bpy)₂ (DIP), M(phen)₂ (DIP) or M(DIP)₂ (phen).

5. The method of claim 4, wherein M is Ru, Rh, or Co.

6. The method of claim 5, wherein M is Ru.

7. The method of claim 5, wherein M is Rh.

8. The method of claim 1, wherein the complex has the formula M(ethylenediamine)₂ (phi);

wherein M is Ru, Rh, or Co.

9. The method of claim 5, wherein the complex has the formula Ru(bpy)₂ (phen)²⁺.

10. The method of claim 5, wherein the complex has the formula Ru(phen)₂ (bpy)²⁺.

11. The method of claim 5, wherein the complex has the formula Ru(phen)₂ (flone)²⁺.

12. The method of claim 5, wherein the complex has the formula Ru(bpy)₂ (DIP)²⁺.

13. The method of claim 5, wherein the complex has the formula Ru(phen)₂ (DIP)²⁺.

14. The method of claim 5, wherein the complex has the formula Ru(DIP)₂ (phen)²⁺.

15. The method of claim 5, wherein the complex has the formula Ru(phi)₂ (bpy)²⁺.

16. The method of claim 5, wherein the complex has the formula Ru(phen)₂ (phi)²⁺.

17. The method of claim 5, wherein the complex has the formula Ru(bpy)₂ (phi)²⁺.

18. The method of claim 5, wherein the complex has the formula Rh(phi)₂ (bpy)³⁺.

19. The method of claim 5, wherein the complex has the formula Rh(phen)₂ (phi)³⁺.

20. The method of claim 5, wherein the complex has the formula Rh(phi)₂ (4,4'"'"' diphenyl bipyridine)³⁺.

21. The method of claim 5, wherein the complex has the formula Rh(bis 4,4'"'"' methyl bipyridylate)₂ (phi)³⁺.

22. The method of claim 5, wherein the complex has the formula Rh(bis 4,4'"'"' bipyridylamide)₂ (phi)³⁺.

23. The method of claim 1, wherein the complex is the optically resolved delta isomer of the complex.

24. The method of claim 1, wherein the complex is the optically resolved lambda isomer of the complex.

25. A labeled DNA molecule produced according to the method of claim 1.

26. A labeled DNA molecule comprising a DNA molecule and a complex having the formula ##STR6## wherein M is Ru, Rh, Co, Fe, Cr, Cu, Zn, Cd, or Pb and each of R₁, R₂ and R₃ is ethylenediamine or a substituted derivative thereof, bipyridine or a substituted derivative thereof, phenanthroline or a substituted derivative thereof, diazafluorene-9-one or a substituted derivative thereof, phenanthrenequinonediimine or a substituted derivative thereof;

wherein R₁ and R₂ are the same and both are different from R₃.

27. A method for labeling DNA with a complex or salt thereof which is spectroscopically or photoactively determinable when bound to DNA which comprises contacting the DNA with the optically resolved delta isomer of a coordination complex or salt thereof having the formula ##STR7## wherein M is Ru or Rh and R is 9,10-phenanthrenequinone diimine or 5-nitro-phenanthroline.

28. A labeled DNA molecule produced according to the method of claim 27.

29. A labeled DNA molecule comprising a DNA molecule and a complex having the formula ##STR8## wherein M is Ru or Rh and R is 9,10-phenanthrenequinone diimine or 5-nitrophenanthroline.

30. A method for selectively labeling a conformation present in a double stranded DNA comprising the conformation which comprises contacting the DNA with a complex or an isomer of the complex having the formula ##STR9## wherein M is Ru, Rh, Co, Fe, Cr, Cu, Zn, Cd, or Pb and each of R₁, R₂ and R₃ is ethylenediamine or a substituted derivative thereof, bipyridine or a substituted derivative thereof, phenanthroline or a substituted derivative thereof, diazafluorene-9-one or a substituted derivative thereof, phenanthrenequinonediimine or a substituted derivative thereof;

wherein R₁, R₂ and R₃ are bound to M by coordination bonds and wherein R₁ and R₂ are the same and both are different from R₃ ;

so that the complex or the isomer binds to the conformation thereby labeling the conformation.

31. A method of claim 30, wherein the conformation is Z-DNA, A-DNA, B-DNA, or a cruciform of DNA.

32. A method for detecting the presence of a conformation present in double stranded DNA which comprises selectively labeling the conformation according to claim 30 and detecting the presence of the complex or the isomer of the complex bound to the conformation.

33. A method of claim 32, wherein the conformation is Z-DNA, A-DNA, B-DNA, or a cruciform of DNA.

34. A method for labeling DNA with a complex or salt thereof which is spectroscopically or photoactively determinable when bound to DNA which comprises contacting the DNA with the optically resolved lambda isomer of a coordination complex or salt thereof having the formula ##STR10## wherein M is Ru or Rh and R is 9,10-phenanthrenequinone diimine or 5-nitro-phenanthroline.