Acyclic nucleoside analogs and oligonucleotide sequences containing them

• US 5,576,427 A
• Filed: 05/26/1995
• Issued: 11/19/1996
• Est. Priority Date: 03/30/1993
• Status: Expired due to Fees

• Current Assignee: Sterling Winthrop, Inc. (GlaxoSmithKline PLC)
• Original Assignee: Sterling Winthrop, Inc. (GlaxoSmithKline PLC)
• Inventors: Guinosso, Charles, Delecki, Daniel J., Cook, Philip D.
• Primary Examiner(s): RORIES, CHARLES
• Application Number:
  US08/452,196
• Time in Patent Office: 543 Days
• Field of Search: 514/44, 536/24.5, 536/24.3, 536/24.31, 536/24.32, 536/23.1, 536/25.3, 544/276, 544/277
• US Class Current: 536/23.1
• CPC Class Codes: C07D 473/18 one oxygen and one nitrogen atom, e.g. guanine
• CPC Class Codes: C07D 473/34 attached in position 6, e.g. adenine
• CPC Class Codes: C07F 9/65616 containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs
• 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: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218

Abstract

Compounds of formula I; ##STR1## wherein R¹ is hydrogen, or a blocking group that is compatible with oligonucleotide synthesis;

R² is hydrogen, nitro, lower alkyl amino, diloweralkyl amino or methyl;

R³ is hydrogen or --P(R⁴)OR⁵ ;

R⁴ is chlorine, 4-nitroimidazole, imidazole, tetrazole, triazole or di(lower-alkyl)amino-;

R⁵ is methyl, 2-cyanoethyl or 2,2,2-trichloroethyl;

n is from 0 to 2

X is oxygen, sulfur, or --NR⁶ ;

R⁶ is hydrogen or lower alkyl;

Q is chosen from the group consisting of ##STR2## R⁷ is lower-alkyl or loweralkyloxy methylene; and R⁸ is hydrogen, benzoyl, anisoyl, or lower-alkyl carbonyl

and its pharmaceutically acceptable addition salts are nucleotide analogs. Modified oligonucleotides containing the nucleoside analogs of formula I are stable to nuclease degradation and are useful in inhibiting gene expression, in sequencing, and in mutagenesis.

Claims

1. A compound of the formula:

##STR19## wherein R¹ is hydrogen, or a blocking group that is compatible with oligonucleotide synthesis;

R² is hydrogen or methyl;

R³ is hydrogen or --P(R⁴)OR⁵ ;

R⁴ is chloro, 4-nitroimidazolyl, imidazolyl, tetrazolyl, triazolyl or di(lower-alkyl)amino-;

R⁵ is methyl, 2-cyanoethyl or 2,2,2-trichloroethyl.

2. A compound according to claim 1, having the formula ##STR20##

3. A compound according to claim 2 wherein R⁵ is cyanoethyl, and R⁴ is diisopropylamino.

4. A compound comprising an oligonucleotide sequence of from about 6 to about 200 bases in which one or more nucleosides at the 3'"'"' terminus of said oligonucleotide are replaced by a residue of formula ##STR21## wherein R² is hydrogen or methyl.

5. A compound according to claim 4 wherein said oligonucleotide sequence comprises from about 12 to about 24 bases.

6. A compound according to claim 5 wherein said oligonucleotide sequence comprises 15 bases.

7. A compound of formula ##STR22## according to claim 6.

8. A method of inhibiting nuclease degradation of an oligonucleotide comprising replacing one or more nucleosides at the 3'"'"' terminus of said oligonucleotide by a residue of formula ##STR23## wherein R₂ is hydrogen, nitro, lower-alkyl amino, diloweralkyl amino or methyl;

X is oxygen, sulfur or --NR⁶ ;

n is 0 to 2;

R⁶ is hydrogen or lower-alkyl;

Q is chosen from the group consisting of ##STR24## R⁷ is lower-alkyl or loweralkyloxy ethylene; and

R⁸ is hydrogen, benzoyl, anisoyl, or lower-alkyl carbonyl.