Stereospecific Grignard-Activated Solid Phase Synthesis of DNA Methylphosphonate Dimers.

Abstract

Stereoregular R(p) or S(p) DNA methylphosphonate dimers have been synthesized on a solid phase support. A deprotected 5'-hydroxyl-N(2)-isobutanoyldeoxyguanosine 3'-O-succinate coupled to high-loaded polyethylene glycol (PEG) coated polystyrene beads (HLP) was activated with a Grignard reagent, t-BuMgCl. After activation was complete, a pure diastereoisomer of 5'-(dimethoxytrityl) N-benzoyldeoxynucleoside 3'-(p-nitrophenyl methylphosphonate) p-nitrophenyl ester (R(p) or S(p)) was added. Coupling of the activated 5'-hydroxyl to the 3'-methylphosphonate ensued, releasing nitrophenol, yielding the R(p) or S(p) dimer, respectively. The dimers were then cleaved from the solid support, deprotected, and purified, yielding methylphosphonate DNA dimers of defined stereochemistry.