Oligo (.alpha.-arabinofuranosyl nucleotides) and .alpha.-arabinofuranosyl precursors thereof

• US 5,177,196 A
• Filed: 08/16/1990
• Issued: 01/05/1993
• Est. Priority Date: 08/16/1990
• Status: Expired due to Fees

• Current Assignee: Microprobe Corporation
• Original Assignee: Microprobe Corporation
• Inventors: Adams, A. David, Meyer, Rich B. Jr., Petrie, Charles R.
• Primary Examiner(s): Rollins, John W.
• Application Number:
  US07/568,391
• Time in Patent Office: 873 Days
• Field of Search: 435/6, 536/27
• US Class Current: 536/22.1
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 19/16 Purine radicals
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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

Abstract

Novel oligonucleotides formed from α-D-arabinofuranosyl nucleoside monomers, including oligonucleotides in which one or more of the monomer units is functionalized, are disclosed herein, as well as functionalized monomeric α-D-arabinofuranosyl nucleosides and nucleotides. A generic formula for the oligomers is: ##STR1## in which B is a nucleotide base which will vary from one monomeric unit to the next in a preselected oligonucleotide sequence; R is phosphate, phsophorothioate, phosphoramidate, or alkanephosphonate; t is 1 for functionalized monomeric units and zero for the others; W is a chemical linker arm; A is a functional group; and n is the number of monomeric units in the oligomer. The oligomers are useful for diagnostic and chemotherapeutic uses. A novel reaction is also disclosed, in which an α-D-arabinofuranosyl nucleoside with exposed hydroxyls at the 2'"'"'- and 3'"'"'-positions is selectively protected at the 2'"'"'-position in a single reaction.

Claims

1. An oligonucleotide comprising a chain of nucleotide units, each said nucleotide unit having the formula ##STR8## in which:

R is a member selected from the group consisting of phosphate, phosphorothioate, phosphoramidate, and alkanephosphonate, and either is the same in all said nucleotide units or varies among said nucleotide units;

B is a nucleotide base, or a nucleotide base bearing a radioactive atom;

W is an alkylene group having 1 to 12 carbon atoms;

an alkenylene group of 2 to 12 carbon atoms and 1 or 2 olefinic bonds;

an alkynylene group of 2 to 12 carbon atoms and 1 or 2 acetylenic bonds;

--O--, --S--, --NH-- --CO--, --NHCO--, --NHCH₂ CO-- or hydroxy-substituted alkylene group having 1 to 12 carbon atoms, --O--, --S--, --NH-- --CO--, --NHCO--, --NHCH₂ CO-- or hydroxy-substituted alkenylene group of 2 to 12 carbon atoms and 1 or 2 olefinic bonds, or --O--, --S--, --NH-- --CO--, --NHCO--, --NHCH₂ CO-- or hydroxy-substituted alkynylene group of 2 to 12 carbon atoms and 1 or 2 acetylenic bonds, said W forming a linking group between a and B wherein the --O--, --S--, --NH-- --CO--, --NHCO--, --NHCH₂ CO-- or hydroxy groups are remote from B;

A is an intercalator, a crosslinking agent, an antibody having a binding site specific for double-stranded DNA, a reporter group, or a biotin residue and A either is the same in all said nucleotide units where t is 1, or varies among said nucleotide units; and

t is zero or 1, and either is the same in all said nucleotide units or varies among said nucleotide units.

2. An oligonucleotide in accordance with claim 1 in which t is 1 on at least one of said nucleotide units.

3. An oligonucleotide in accordance with claim 1 in which t is zero on all of said nucleotide units.

4. An oligonucleotide in accordance with claim 1 in which R is monophosphate.

5. An oligonucleotide in accordance with claim 1 in which W includes an aliphatic chain of at least three carbon atoms in length.

6. An oligonucleotide in accordance with claim 1 in which W is an aliphatic chain of at least three carbon atoms in length, terminating with a nucleophilic group at the end adjacent to A.

7. An oligonucleotide in accordance with claim 1 in which W is an aliphatic chain of at least three carbon atoms in length, terminating with an amino or an amido group at the end adjacent to A.

9. An oligonucleotide in accordance with claim 1 in which B is a member selected from the group consisting of adenine, guanine, thymine, cytosine and uracil.

10. An oligonucleotide in accordance with claim 1 in which B is a member selected from the group consisting of:

purines, at whose 8-position W is joined in nucleotide units where t is 1,

pyrimidines, at whose 5-position W is joined in nucleotide units where t is 1,

3-deazapurines, at whose 3-position W is joined in nucleotide units where t is 1,

7-deazapurines, at whose 7-position W is joined in nucleotide units where t is 1, and

pyrazolo{3,4-d}pyrimidines, at whose 3-position W is joined in nucleotide units where t is 1.

11. An oligonucleotide in accordance with claim 1 in which B is a member selected from the group consisting of:

adenine, at whose 8-position W is joined in nucleotide units where t is 1;

guanine, at whose 8-position W is joined in nucleotide units where t is 1;

thymine, and t is zero in all nucleotide units where B is thymine;

cytosine, at whose 5-position W is joined in nucleotide units where t is 1; and

uracil, at whose 5-position W is joined in nucleotide units where t is 1.

12. An oligonucleotide in accordance with claim 1 in which the number of nucleotide units in said chain is from about 5 to about 2000.

13. An oligonucleotide in accordance with claim 1 in which the number of nucleotide units in said chain is from about 10 to about 100.

14. An oligonucleotide in accordance with claim 1 in which A is a member selected from the group consisting of intercalators, signal-producing reporter groups, and ligands capable of binding to signal-producing anti-ligands.

8. An oligonucleotide in accordance with claim l..in which B is a member selected from the group consisting of purines, pyrimidines, 3-deazapurines, 7-deazapurines, and pyrazolo{3,4-d}pyrimidines.