Thiol-derivatized oligonucleosides

• US 5,852,182 A
• Filed: 06/02/1995
• Issued: 12/22/1998
• 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:
  US08/458,396
• Time in Patent Office: 1,299 Days
• Field of Search: 536/22.1, 536/23.1, 536/25.3, 536/25.32, 536/5, 536/24.31, 536/27.6, 536/27.8, 536/27.81, 536/28.1, 536/28.4, 536/28.5, 536/28.53, 536/28.54, 536/55.3, 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: 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 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/3523 Allyl
• CPC Class Codes: C12N 2310/3527 Other alkyl chain

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 comprises a ribofuranosyl sugar portion and a base portion; and

at least one of said nucleosides bears at a 2'"'"'-O-position, a 3'"'"'-O-position, or a 5'"'"'-O-position a substituent having formula;

space="preserve" listing-type="equation">--R.sub.s --S--R.sub.1

where;

R_s has formula R_A, R_A --C(O)--Q--R_A, R_A --Q--R_A --Q--R_A ;

each R_A is independently selected from alkyl having from 1 to about 10 carbon atoms;

each Q is, independently, NH, O or S;

R₁ is H or a thiol protecting group or has formula

space="preserve" listing-type="equation">S--R.sub.2, CH.sub.2 C(O)--NH--R.sub.2, CH.sub.2 --CH=CH--C(O)--R.sub.2, --CH.sub.2 --CH.sub.2 --NH--S(O).sub.2 --R.sub.2,

or (maleimido)--R₂ ; and

R₂ comprises a steroid molecule, a reporter molecule, a lipophilic molecule, a reporter enzyme, a peptide, a protein, a reporter group, an alkylator, an intercalator, a cell receptor binding molecule, a crown ether, a crown amine, a porphyrin, a crosslinking agent, a peptide nucleic acid, or a thiol attached to a poly(ethylene glycol).

2. The compound of claim 1 wherein more than one of said nucleosides bears said substituent at a 2'"'"'-O-position, a 3-O-position, or a 5'"'"'-O-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 1 wherein R₁ is S--R₂ and R₂ is thiocholesterol.

7. The compound of claim 1 wherein R₁ is CH₂ C(O)--NH--R₂ and R₂ is o-phenanthroline.

8. The compound of claim 1 wherein R₁ is phospholipid maleimido, fluorescein maleimido, or pyrene maleimido.