Chemical probes for left-handed DNA and chiral metal complexes as Z-specific anti-tumor agents

• US 4,721,669 A
• Filed: 01/18/1985
• Issued: 01/26/1988
• 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): Kepplinger, Esther M.
• Assistant Examiner(s): Jay, Jeremy
• Application Number:
  US06/693,023
• Time in Patent Office: 1,103 Days
• Field of Search: 546/4, 546/10, 546/88, 536/22, 536/27, 556/136, 556/138, 424/1.1, 424/9, 424/131, 424/149, 435/6, 935/77, 935/78, 204/157.71, 204/157.72
• US Class Current: 435/6.12
• CPC Class Codes: C07D 471/04 Ortho-condensed systems
• CPC Class Codes: C07F 15/065 without a metal-carbon linkage
• 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 and salts and optically resolved enantiomers thereof, of the formula (R)₃ --M, wherein R comprises 1,10-phenanthroline or a substituted derivative thereof, M comprises a suitable transition metal, e.g. ruthenium(II) or cobalt(III), and R is bonded by M by a coordination bond.

The complexes of this invention are useful in methods for labeling, nicking and cleaving DNA. The lambda enantiomer of complexes of this invention is useful in methods for specifically labeling, detecting, nicking and cleaving Z-DNA.

The complexes may also be used in a method for killing tumor cells and may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition for the treatment of tumor cells in a subject. The invention further concerns methods for treating a subject afflicted with tumor cells.

Claims

1. A method for detecting in a sample the presence of double-stranded DNA comprising contacting the sample with a complex of the formula (R)₃ --Co (III), wherein R comprises 1,10-phenanthroline or a substituted derivative thereof and R is bound to Co(III) by a coordination bond, the contacting being effected under conditions such that the complex binds to double-stranded DNA present in the sample by intercalation and thereby labels the double-stranded DNA with the complex, detecting DNA labeled with the complex, and thereby detecting the presence of double-stranded DNA in the sample.

2. A method of claim 1, wherein the substituted derivative of 1,10-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;

3,8-diphenyl-1,10-phenanthroline;

4,7-dispermine-1,10-phenanthroline, or 3,8-dispermine-1,10-phenanthroline.

3. A method for detecting in a sample the presence of double-stranded Z-DNA which comprises contacting the sample with the lambda enantiomer of a complex having the formula (R)₃ --M, wherein R comprises 1,10-phenanthroline or a substituted derivative thereof, M comprises ruthenium (II) or cobalt (III) and R is bound to M by a coordination bond, the contacting being effected under conditions such that the complex binds to double-stranded Z-DNA present in the sample by intercalation and thereby labels the double-stranded Z-DNA with the lambda enantiomer of the complex, detecting Z-DNA labeled with the lambda enantiomer of the complex, and thereby detecting the presence of double-stranded Z-DNA in the sample.

4. A method of claim 3 wherein M is ruthenium(II).

5. A method of claim 3, wherein M is cobalt(III).

6. A method of claim 3, wherein the substituted derivative of 1,10 -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;

3,8-diphenyl-1,10-phenanthroline;

4,7-dispermine-1,10-phenanthroline, or 3,8-dispermine-1,10-phenanthroline.

7. A method for nicking double-stranded DNA by effecting breakage of at least one phosphodiester bond along the DNA which comprises contacting the DNA with a coordination complex or salt thereof having the formula (R)₃ --Co (III), wherein R comprises 1,10-phenanthroline or a substituted derivative thereof and R is bound to Co by a coordination bond, under conditions such that the complex binds to the DNA by intercalation to form an adduct and irradiating the adduct so formed with ultraviolet radiation of a wavelength which is absorbed by the ligand bands of the complex so as to nick the DNA at the site(s) of intercalation.

8. A method for cleaving double-stranded DNA which comprises nicking the DNA according to claim 7 and treating the nicked DNA so produced with an enzyme which is not deactivated in the presence of the complex used for DNA nicking and capable of cleaving single-stranded DNA so as to cleave the nicked DNA at the site of the nick(s).

9. A method for selectively nicking Z-DNA, present in DNA comprising Z-DNA and other forms of DNA, by effecting breakage of at least one phosphodiester bond along the Z-DNA which comprises contacting the DNA containing the Z-DNA with a Λ

isomer of a coordination complex or salt thereof having the formula (R)₃ --CO(III), wherein R comprises 1,10-phenanthroline or a substituted derivative thereof and R is bound to Co by a coordination bond, under conditions such that the Λ

isomer binds to the DNA by intercalation to form an adduct and irradiating the adduct so formed with ultraviolet radiation of a wavelength which is absorbed by the ligand bands of the Λ

isomer so as to nick the DNA at the site(s) of intercalation.

10. A method for selectively cleaving double-stranded Z-DNA, present in DNA comprising Z-DNA and other forms of DNA, which comprises nicking the Z-DNA according to claim 9 and treating the nicked DNA so produced with an enzyme which is not deactivated in the presence of the complex used for DNA nicking and capable of cleaving single-stranded DNA so as to cleave the nicked DNA at the site of the nick(s).