Substituted 2-nitrobenzyltrichloroacetate esters for photodirected oligonucleotide detritylation in solid films.
Oligonucleotide microarray fabrication by chemical synthesis using photoacid generators in solid films could have advantages over existing methods, but has not matched the accuracy of conventional synthesis where detritylation is performed with acid solutions. To address this problem, we explored the kinetics and equilibria of nucleoside detritylation in solid films, using trichloroacetic acid (TCA) generated by photolysis from its esters with substituted 2-nitrobenzyl alcohols. We synthesised 25 such esters, all alpha-phenyl substituted, and assessed their potential as solid film photoacid generators. They included sets with (i) mono- or dimethoxy-, (ii) 5-halo-, (iii) alkyl- or aryl-substituted 5-amino-, or (iv) 5-aryl-substituents in the 2-nitro- or 2,6-dinitrobenzyl ring. Absorption maxima of their UV spectra ranged from 230 to 410 nm, with quantum yields at 365 nm from < 0.01 to nearly 1.0. The esters formed optically clear solid films on glass slides without added polymer. Kinetics of intrafilm photoacid generation, proton activity changes and detritylation were measured in situ. The most effective esters for light sensitivity and detritylation were 5-chloro-, 5-bromo-, 4,5-dimethoxy-, and 4- or 5-aryl-substituted 2,6-dinitrobenzyl esters. Photoacid-induced increases in proton activity and detritylation were severely inhibited by polymers containing electronegative heteroatoms, but not by polymers lacking them. In solid films, intrafilm detritylation with photogenerated TCA was fast, but stopped at an equilibrium well short of completion. Both experiment and theory emphasise the inadequacy of attempting to force detritylation with high intrafilm acid activity.
Serafinowski
Pawel J
PJ
Cancer Research UK Centre for Cancer Therapeutics, Sutton, Surrey, UK SM2 5NG. pawel.serafinowski@icr.ac.uk
Garland
Peter B
PB
D000085:Acetates / Q000737:chemistry*; D000143:Acids / Q000737:chemistry; D004952:Esters / Q000138:chemical synthesis / Q000737:chemistry*; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015394:Molecular Structure; D009578:Nitrobenzenes / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D010777:Photochemistry; D017319:Photosensitizing Agents / Q000138:chemical synthesis* / Q000737:chemistry; D012996:Solutions; D013053:Spectrophotometry
D000085:Acetates; D000143:Acids; D004952:Esters; D009578:Nitrobenzenes; D009841:Oligonucleotides; D017319:Photosensitizing Agents; D012996:Solutions; C006972:chloroacetic acid; C036077:nitrobenzene