Thiol-derivatized nucleosides and oligonucleosides

• US 6,265,558 B1
• Filed: 08/26/1999
• Issued: 07/24/2001
• Est. Priority Date: 01/11/1990
• Status: Expired due to Term

• Current Assignee: ISIS Pharmaceuticals Incorporated
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Manoharan, Muthiah, Cook, Phillip Dan
• Primary Examiner(s): Wilson, James O.
• Application Number:
  US09/383,856
• Time in Patent Office: 698 Days
• Field of Search: 536/23.1, 536/22.1, 536/24.31, 536/25.3, 536/25.32, 536/27.6, 536/27.8, 536/27.81, 536/28.1, 536/28.4, 536/28.5, 530/395
• US Class Current: 536/23.1
• 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 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• 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
• CPC Class Codes: C07J 43/003 not condensed
• CPC Class Codes: C07J 51/00 Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
• 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
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/3125 Methylphosphonates
• 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 2310/333 Modified A
• CPC Class Codes: C12N 2310/336 : Modified G
• 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 2310/3531 : Hydrogen
• CPC Class Codes: C12N 2310/3533 : Halogen
• CPC Class Codes: C12N 9/0069 : acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• CPC Class Codes: C12Y 113/11012 : Linoleate 13S-lipoxygenase (1.13.11.12)

Abstract

Nucleosides and linked nucleosides functionalized to include alkylthiol chemical functionality at ribofuranosyl positions, nucleosidic base positions, or on internucleoside linkages. In certain embodiments, the compounds of the invention further include steroids, reporter molecules, reporter enzymes, lipophilic molecules, peptides or proteins attached to the nucleosides through the alkylthio group.

Claims

1. A compound comprising a plurality of linked nucleosides, wherein:

each nucleoside includes a ribofuranosyl sugar portion and a base portion; and

at least one of said nucleosides bears at a 5-pyrimidine position or at a 2-, 6-, or 8-purine position a substituent having formula;

2. The compound of claim 1 wherein more than one of said nucleosides bears said substituent at a 5-pyrimidine position or at a 2-, 6-, or 8-purine position.

3. The compound of claim 1 wherein R_A is (CH₂)_n where n is an integer from 1 to about 10.

4. The compound of claim 3 wherein n is 6.

5. The compound of claim 1 wherein R₁ is H or trityl.

6. The compound of claim 3 wherein said nucleoside bears at a 5-pyrimidine position a substituent having formula —

C≡

C—

CH₂—

NH—

C(O)—

(CH₂)_n—

S—

R₁.

7. The compound of claim 3 wherein said nucleoside bears at a 5-pyrimidine position a substituent having formula —

CH═

CH—

C(O)—

NH—

(CH₂)_n—

S—

R₁.

8. The compound of claim 3 wherein said nucleoside bears at a 2- or 8-purine position a substituent having formula —

NH—

(CH₂)_n—

S—

R₁.

9. A compound comprising a plurality of linked nucleosides, wherein:

each nucleoside includes a ribofuranosyl sugar portion and a base portion; and

at least two of said nucleosides are bound by a linkage having formula;

10. The compound of claim 9 wherein R_A is (CH₂)_n where n is an integer from 1 to about 10.

11. The compound of claim 10 wherein n is 6.

12. The compound of claim 10 wherein R₁ is H or trityl.

13. The compound of claim 10 wherein said nucleosides are bound by a linkage having formula —

O—

CH₂—

P[O][S—

(CH₂)_n—

S—

R₁]—

O—

.