Nucleic acid intercalating agents

• US 5,175,273 A
• Filed: 07/01/1988
• Issued: 12/29/1992
• Est. Priority Date: 07/01/1988
• Status: Expired due to Term

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Bischofberger, Norbert W., Matteucci, Mark D.
• Primary Examiner(s): Brown, Johnnie R.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US07/213,957
• Time in Patent Office: 1,642 Days
• Field of Search: 536/27, 536/28, 536/29, 536/1.1, 536/18.7, 536/22, 536/23, 435/6
• US Class Current: 536/26.13
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• 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: C12Q 1/6832 Enhancement of hybridisation reaction

Abstract

Pyridinone or pyrimidinone nucleoside bases containing fused aromatic polycyclic rings are provided. These polycyclic nucleosides are incorporated into oligonucleotides and hybridized to complementary nucleic acid. Fluorescence spectroscopy and thermal denaturation profiles provided evidence that the polycyclic base is intercalated into the resulting duplex. The fused polycyclic ring systems optionally are substituted with reactive species which inactivate complementary nucleic acids. The oligonucleotides of this invention are useful as improved probes, diagnostic reagents, or for cleaving or derivatizing predetermined domains within nucleic acids.

Claims

1. A pyridinone or pyrimidinone nucleoside comprising a fused aromatic polycyclic base having molecular dimensions equal to or less than about 30-50 angstroms by about 30-50 angstroms by about 3-7 angstroms, wherein the nucleoside is represented by the structure ##STR18## wherein R₃ is an aromatic polycycle comprising up to four fused aromatic rings;

Y is C or N;

R₇ is N or ═

C(R₁)--;

R₁ and R₆ are independently selected from the group consisting of H, halogen, C₁ -C₁₀ -alkyl, saturated or unsaturated cycloalkyl, C₁ -C₁₀ -alkylcarbonyloxy, C₁ -C₁₀ -alkyl, hydroxy-C₁ -C₁₀ -alkyl, heterocycle (N,O, or S), and nitro and;

R₂ is a ribose or deoxyribose sugar.

2. The nucleoside of claim 1 wherein Y is N.

3. The nucleoside of claim 2 wherein R₃ is selected from the group of ##STR19## wherein R₈ =CH₃, NH₂, N(CH₃)₂, RCO, OH, C₁ -C₁₀ -alkoxy, C₁ -C₁₀ -alkyl, C₆ -C₁₂ -aryl, ethylenediamine tetraacetic acid, or thioether, R₉ =S, and Z is O or S.

4. The nucleoside of claim 1 wherein R₃ is the residue of an aromatic polycyclic ring containing from 2 to 4 cycloalkylene rings.

5. The nucleoside of claim 1 wherein the aromatic polycycle is completely unsaturated.

6. The nucleoside of claim 1 wherein the aromatic polycycle contains one or more substituents selected from the group oxo, hydroxyl, thio-C₁ -C₁₀ -alkyl, amino, C₁ -C₁₀ -alkylamino, formyl, C₁ -C₁₀ -alkoxy, C₁ -C₁₀ -alkylcarbonyl, C₆ -C₁₂ -aryl, and ethylenediamine tetraacetic acid.

7. The nucleoside of claim 1 wherein the aromatic polycycle comprises a heterocyclic ring containing O, S or N.

8. The nucleoside of claim 1 wherein the aromatic polycycle comprises a quinone.

9. The nucleoside of claim 8 wherein the aromatic polycycle comprises an anthraquinone.

10. The nucleoside of claim 4 wherein the cycloalkylene rings contain from 4 to 7 carbon atoms.

11. The nucleoside of claim 1 wherein R₁ is cyclic or acyclic alkenyl, or halogen.

12. The nucleoside of claim 1 wherein R₁ is methyl, C₁ -C₁₀ -alkoxy, or hydrogen.

13. An oligonucleotide of the structure:

##STR20## wherein A is an insoluble matrix or the nucleoside ##STR21## R₅ is H or hydroxyl;

R₄ is O, S, alkyl, alkylamine, or alkyl ether;

n is an integer; and

B is a nucleoside base;

wherein at least one base is of the structure;

##STR22## and is positioned within about the first 20 percent or about the last 20 percent of the length of the oligonucleotide, and wherein

R₃ is the residue of an aromatic polycycle;

Y is C or N;

R₇ is N or ═

C(R₁)--;

R₁ and R₆ are independently selected from the group consisting of hydrogen, halogen, C₁ -C₁₀ -alkoxy, saturated or unsaturated cycloalkyl, C₁ -C₁₀ -alkylcarbonyloxy, C₁ -C₁₀ -alkyl, hydroxy-C₁ -C₁₀ -alkyl, heterocycle (N,O,S), and nitro.

14. The oligonucleotide of claim 13 wherein the bases are guanidine, cytosine, adenosine, uracil, or thymine.

15. The oligonucleotide of claim 13 wherein n ranges about from 5 to 70.

16. The oligonucleotide of claim 13 wherein the polycyclic aromatic base is present in the second nucleoside 5'"'"' or 3'"'"' from the end of the oligonucleotide.

17. The oligonucleotide of claim 13 wherein the polycyclic aromatic base is fluorescent.

18. The oligonucleotide of claim 13 wherein the polycyclic aromatic base is ##STR23##

19. The compound of claim 13 wherein R₃ is in substantially the same plane as the group ##STR24##