2'deoxy-2'-alkylnucleotide containing nucleic acid

• US 5,627,053 A
• Filed: 05/02/1995
• Issued: 05/06/1997
• Est. Priority Date: 03/29/1994
• Status: Expired due to Term

• Current Assignee: Ribozyme Pharmaceuticals, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Ribozyme Pharmaceuticals, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: Beigelman, Leonid, Karpeisky, Alexander, Usman, Nassim, Modak, Anil
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/432,874
• Time in Patent Office: 735 Days
• Field of Search: 435/6, 435/5, 435/91.1, 514/44, 536/24.5
• US Class Current: 435/91.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• 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: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: C12N 15/1131 against viruses
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/121 Hammerhead
• CPC Class Codes: C12N 2310/122 Hairpin
• CPC Class Codes: C12N 2310/123 Hepatitis delta
• CPC Class Codes: C12N 2310/1241 Tetrahymena
• CPC Class Codes: C12N 2310/126 involving RNAse P
• CPC Class Codes: C12N 2310/127 DNAzymes
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/32 : of the sugar
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 : 2'-R Modification
• CPC Class Codes: C12N 2310/33 : of the base
• CPC Class Codes: C12N 2310/332 : Abasic residue
• CPC Class Codes: C12N 2310/333 : Modified A
• CPC Class Codes: C12N 2310/334 : Modified C
• CPC Class Codes: C12N 2310/335 : Modified T or U
• CPC Class Codes: C12N 2310/336 : Modified G
• CPC Class Codes: C12N 2310/346 : having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3513 : Protein; Peptide
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12N 2310/3523 : Allyl
• CPC Class Codes: C12N 2310/3527 : Other alkyl chain
• CPC Class Codes: C12N 2310/3533 : Halogen
• CPC Class Codes: C12N 2310/3535 : Nitrogen
• CPC Class Codes: C12Y 304/24017 : Stromelysin 1 (3.4.24.17)

Abstract

2'"'"'-deoxy-2'"'"'-alkylnucleotides useful for stabilizing enzymatic nucleic acid molecules and antisense molecules.

Claims

1. Oligonucleotide comprising one or more 2'"'"'-deoxy-2'"'"'-alkyl-nucleotides.

2. Enzymatic nucleic acid Capable of cleaving single-stranded RNA or DNA comprising a 2'"'"'-deoxy-2'"'"'-alkylnucleotide.

3. Method for producing an enzymatic nucleic acid molecule having enzymatic activity capable of cleaving a single-stranded RNA or DNA molecule, comprising the step of forming said enzymatic molecule with at least one nucleotide having at its 2'"'"'-position an alkyl group.

4. 2'"'"'-deoxy-2'"'"'-alkylnucleotide triphosphate.

5. Method for synthesis of a 2'"'"'-C-allyl derivative of a nucleoside from a 5'"'"'-O-dimethoxytrityl-3 '"'"'-O-t-bulydimethylsilyl-base comprising the steps of:

(a) Phenoxythiocarbonylation of 5'"'"'O-dimethoxytrityl-3'"'"'-O-t-bulydimethylsilyl-base to yield a thioester, replacing a 2'"'"' hydroxyl group with a phenoxythiocarbonyl groups, and

(b) Heck allylation of said thioester to form a 2'"'"'-C-allyl derivative in which said 2'"'"'-phenoxythiocarbonyl group is replaced with a 2'"'"'-allyl group to yield said 2'"'"'-C-allyl derivative.