Oligonucleotides containing 4'-substituted nucleotides

• US 5,446,137 A
• Filed: 12/09/1993
• Issued: 08/29/1995
• Est. Priority Date: 12/09/1993
• Status: Expired due to Term

• Current Assignee: DADE Behring Beteiligungs GmbH (Siemens AG)
• Original Assignee: Syntec Incorporated (Atos SE)
• Inventors: Maag, Hans, Rose, Samuel J., Schmidt, Beat
• Primary Examiner(s): Stone, Jacqueline M.
• Assistant Examiner(s): CAMPELL, BRUCE R
• Application Number:
  US08/164,893
• Time in Patent Office: 628 Days
• Field of Search: 514/44, 536/23.1, 536/24.3, 536/24.5, 536/25.3, 536/25.32
• US Class Current: 536/23.1
• 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

Abstract

This invention relates to oligonucleotides having at least one nucleotide that is substituted at the 4'"'"' position of the sugar moiety with a substituent other than hydrogen. These oligonucleotides are useful in hybridization assays and as therapeutic agents.

Claims

1. An oligonucleotide which comprises at least one nucleotide having the structure:

##STR15## wherein;

A is a purine or a pyrimidine;

B and B'"'"' are selected from the group consisting of H, OH, F, OCH₃ and SCH₃, provided that at least one is H;

D'"'"' is selected from the group consisting of OH, O--P(O)(OH)--OX, O--P(S)(OH)--OX, O--P(S)(SH)--OX and O--P(O)(CH₃)--OX, where X is selected from the group consisting of H, a nucleotide, and a protecting group;

E'"'"' is RY, where Y is selected from the group consisting of H and a substituent that renders said nucleotide modifiable, separable or detectable and R is a linking group; and

F is selected from the group consisting of OH and O--P(O)(OH)--OX, where X is selected from the group consisting of H, a nucleotide and a protecting group.

2. The oligonucleotide of claim 1 wherein R is selected from the group consisting of alkylene, alkyl, alkenyl, alkadienyl, aralkyl, aminoalkyl and alkylether.

3. The oligonucleotide of claim 1 wherein said protecting group is selected from the group consisting of phthalyl, carbobenzyloxy, benzyl, benzoyl, trityl, monomethoxytrityl, dimethoxytrityl, acetyl, trifluoroacetyl, trimethylsilyl, t-butyl(dimethyl)silyl, t-butyl(diphenyl) silyl, carbonate, 2-trimethylsilylethyl, methoxymethyl, 2-methoxyethoxymethyl and dihydropyranyl.

4. The oligonucleotide of claim 1 wherein Y is biotin.

5. The oligonucleotide of claim 1 wherein Y is a reactive functional group that renders said oligonucleotide modifiable and is selected from the group consisting of NH₂, SH, OH, COOH, NHC(O)CF₃ and CHO.

6. The oligonucleotide of claim 1 wherein Y is a substituent that renders said oligonucleotide modifiable and is selected from the group consisting of nucleic acid modifying catalysts, DNA intercalators, minor groove binders and cleavage agents.

7. The oligonucleotide of claim 6 wherein said nucleic acid modifying catalyst is selected from the group consisting of nucleases, methylases and metal chelates.

8. The oligonucleotide of claim 6 wherein said DNA intercalator is selected from the group consisting of ethidium, ethidium bromide, acridines, phenazine, phenazinium salts, pyrenes, diazapyrenes, psoralen, psoralen analogs and anthraquinones.

9. The oligonucleotide of claim 6 wherein said minor groove binder is selected from the group consisting of distamycin, netropsin and bis-benzimidazoles.

10. The oligonucleotide of claim 6 wherein said cleavage agent is selected from the group consisting of metal complexes and DNA intercalators.

11. The oligonucleotide of claim 1 wherein Y is selected from the group consisting of detectable labels and specific binding pair members.

12. The oligonucleotide of claim 11 wherein said detectable label is selected from the group consisting of enzymes, fluorescers, chemiluminescers, sensitizers, particles and catalysts.

13. The oligonucleotide of claim 1 wherein A is selected from the group consisting of adenine, guanine, xanthine, hypoxanthine, uracil, thymine and cytosine.

14. The oligonucleotide of claim 1 wherein A is selected from the group consisting of 5-(1-propynyl)pyrimidine, 5-halopyrimidines and 7-deaza-7-alkylpurines.

15. A method of producing an oligonucleotide, which comprises the step of incorporating into said oligonucleotide at least one nucleotide having a substituent other than hydrogen at the 4'"'"' position of the sugar moiety, said substituent producing, or being capable of producing, the alteration of said oligonucleotide.

16. The method of claim 15 wherein said incorporating involves the step of synthesizing said oligonucleotide using a nucleotide precursor having a substituent other than hydrogen at the 4'"'"' position of the sugar moiety.

17. The method of claim 15 wherein said altering further comprises the step of hybridizing to said oligonucleotide a polynucleotide that lacks a nucleotide having a 4'"'"'-substituent other than hydrogen.