Method of making oligonucleotides and oligonucleotide analogs using phospholanes and enantiomerically resolved phospholane analogues

• US 5,646,267 A
• Filed: 06/05/1995
• Issued: 07/08/1997
• Est. Priority Date: 08/05/1991
• Status: Expired due to Fees

• Current Assignee: Polish Academy of Sciences (Government of Poland)
• Original Assignee: Polish Academy of Sciences (Government of Poland)
• Inventors: Stec, Wojciech J., Grajkowski, Andrzej, Uznanski, Bogdan
• Primary Examiner(s): Kight, John
• Assistant Examiner(s): CRANE, LAWRENCE E
• Application Number:
  US08/462,418
• Time in Patent Office: 764 Days
• Field of Search: 536/25.33, 536/25.1, 536/25.2, 536/25.3, 536/23.1
• US Class Current: 536/25.33
• CPC Class Codes: C07F 9/6564 having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
• CPC Class Codes: C07F 9/6578 having phosphorus and sulfur atoms with or without oxygen atoms, as ring hetero atoms
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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

Abstract

Methods and compounds are provided for solid phase synthesis of oligonucleotides and related polymers by condensing protected monomer-O-[1,3,2-dichalcogen-substituted-phospholane] synthons in the presence of a catalytic base. Compounds of the invention include 2-N-substituted-1,3,2-dichalcogen-substituted-phospholane precursors of the above synthons, the protected monomer-O-[1,3,2-dichalcogen-substituted-phospholane] synthons, and P-chiral oligonucleotides and related P-chiral polymers.

Claims

1. A method of synthesizing a P-chiral oligonucleotide phosphorothioate of a predetermined length, the P-chiral oligonucleotide comprising a sequence of nucleosides or nucleoside analogs and phosphorothioate or phosphodiester linkages, the phosphorothioate linkages being R_p chiral or S_p chiral, the method comprising the steps of:

(a) providing a first nucleoside or nucleoside analog attached to a solid phase support, the first nucleoside or nucleoside analog having a protected hydroxyl;

(b) deprotecting the protected hydroxyl to form a free hydroxyl;

(c) reacting with the free hydroxyl a synthon selected from the group defined by the formula;

##STR9## wherein;

R^I is a hydroxyl protecting group;

B is a purine or a pyrimidine;

D is hydrogen, halogen, hydroxyl, or --OR'"'"', wherein R'"'"' is alkyl having from 1 to 3 carbon atoms or a 2'"'"'-hydroxyl protecting group;

D'"'"' is hydrogen, hydroxyl, or --OR" such that R" is a 3'"'"'-hydroxyl protecting group;

X is sulfur, Y is oxygen, and Z is sulfur; and

R³, R⁴, R⁵, and R⁶ are separately hydrogen or alkyl having from 1 to 4 carbon atoms or are separately or together with the carbon atoms to which they are attached aryl or alkyl-substituted aryl having from 6 to 12 carbon atoms; and

(d) repeating steps (b) and (c) until the predetermined length of the P-chiral oligonucleotide phosphorothioate is achieved.

2. The method of claim 1 wherein said step of reacting includes reacting in the presence of a catalytic base.

3. The method of claim 2 further including the step of capping unreacted free hydroxyls after said step (c).

4. The method of claim 3 wherein said synthon is selected from the group defined by the formula:

##STR10##

5. The method of claim 4 wherein R³, R⁴, R⁵, and R⁶ are separately hydrogen or methyl.

6. The method of claim 5 wherein said predetermined length of said P-chiral oligonucleotide phosphorothioate is in the range of 12 to 60 nucleosides or nucleoside analogs.

7. The method of claim 6 wherein said catalytic base is 1,8-diazabicyclo[5.4.0]undec-7-ene.