Sugar modified nucleosides and their use for synthesis of oligonucleotides

• US 5,712,378 A
• Filed: 11/02/1995
• Issued: 01/27/1998
• Est. Priority Date: 11/02/1994
• Status: Expired due to Fees

• Current Assignee: Marina Biotech, Inc. (Pentair Plc (Ireland))
• Original Assignee: ICN Pharmaceuticals, Inc (Bausch Health Cos., Inc.)
• Inventors: Wang, Guangyi
• Primary Examiner(s): Marschel, Ardin H.
• Assistant Examiner(s): Riley, Jezia
• Application Number:
  US08/552,363
• Time in Patent Office: 817 Days
• Field of Search: 435/6, 536/27.1, 536/23.1, 536/24.3, 536/25.3, 536/25.32
• US Class Current: 536/22.1
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 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

A number of modified nucleosides are disclosed composed of modified sugar moieties which contain substituents at C1 and C4 positions, or branched substituents at C3 and C5 positions of deoxyribose or ribose. Each nucleoside is converted to or properly protected and then converted to the corresponding phosphoramidites. These phosphoramidites are used to assemble oligonucleotides in which there is at least one of the forenoted nucleosides. These sugar modified oligonucleotides have the potential to be used as antisense therapies since they are expected to enhance nuclease resistance and cellular uptake while they maintain sequence-specificity and affinity to nucleic acid targets in vitro or in vivo.

Claims

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

##STR6## wherein;

R₁ is selected from the group consisting of substituted alkyl, aralkyl, substituted aralkyl, aryl, and substituted alkyl;

R₂ is selected from the group consisting of H, OH, alkoxy, aralkoxy and aryloxy;

R₃ and R₄ are independently selected from the group consisting of OH, internucleotide linkage and a hydroxyl blocking group;

X is selected from the group consisting of O and CH₂ ;

B is a nucleoside base selected from the group consisting of Adenine, Guanine, Cytosine Uracil and Thymine;

wherein;

the substituted portion of the substituted alkyl is other than OH when X is O;

any alkyl portion of R₁ and R₂ is C1 to C10 linear or branched, saturated or unsaturated; and

any aryl portion of R₁ and R₂ is a phenyl, polycyclic ring or heterocycle.

2. The oligonucleotide of claim 1, wherein the substituted portion of at least one of the substituted alkyl, substituted aralkyl and substituted aryl is selected from the group consisting of NH₂, NHR'"'"', NR'"'"'R" and +NR'"'"'R"R'"'"'" where R'"'"', R" and R'"'"'" are independently selected from the group consisting of lower alkyl and lower alkylcarbonyl.

3. The oligonucleotide of claim 1, wherein the substituted portion of at least one of the substituted alkyl, substituted aralkyl and substituted aryl is selected from the group consisting of CN, NO₂, N₃, halogen, OR'"'"', SH and SR'"'"' where R'"'"' is selected from the group consisting of lower alkyl and lower alkylcarbonyl.

4. The oligonucleotide of claim 1, wherein the substituted portion of at least one of the substituted alkyl substituted aralkyl and substituted aryl is selected from the group consisting of COOH, COOR'"'"' and CONR'"'"'R" where R'"'"' and R" are independently selected from the group consisting of lower alkyl, aralkyl and aryl.

5. The oligonucleotide of claim 1, wherein the substituted alkyl, substituted aralkyl and substituted aryl independently comprise a linker which is attached to at least one of a functional moiety, an artificial nuclease, a cross-linking reagent, an intercalator, and a reporter molecule.

6. The oligonucleotide of claim 1, wherein X is CH₂ ;

R₁ is selected from the group consisting of alkyl, aralkyl, aryl, hydroxyalkyl, hydroxyaralkyl, and hydroxyaryl.