초록 ▼

The present invention provides synthetic processes by which oligomeric compounds, having at least one phosphodiester or phosphorothioate linkage, are prepared. The synthetic processes use a novel mixture of concentrated ammonium hydroxide and a thiol compound during the deblocking step of all or sel

The present invention provides synthetic processes by which oligomeric compounds, having at least one phosphodiester or phosphorothioate linkage, are prepared. The synthetic processes use a novel mixture of concentrated ammonium hydroxide and a thiol compound during the deblocking step of all or selected internucleoside linkages. Alternatively, the deblocking is carried out in two steps using a mercapto compound in an aqueous amine followed by concentrated ammonium hydroxide. Also provided are synthetic intermediates useful in such processes. Novel oxidation procedures to give phosphodiester oligomers are also included.

대표청구항 ▼

[ What is claimed is:] [1.] A method of preparing an oligomeric compound having at least one moiety of formula: [ STR 8 ] rein:X.sub.1 is O or S;Bx is a heterocyclic base moiety; andR.sub.1 is H, a protected hydroxyl group, a 2'-substituent group, or a protected 2'-substituent group; comprising the

[ What is claimed is:] [1.] A method of preparing an oligomeric compound having at least one moiety of formula: [ STR 8 ] rein:X.sub.1 is O or S;Bx is a heterocyclic base moiety; andR.sub.1 is H, a protected hydroxyl group, a 2'-substituent group, or a protected 2'-substituent group; comprising the steps of:(a) selecting a compound of formula: [ STR 9 ] rein:Bx is a heterocyclic base moiety;R.sub.1 is H, a protected hydroxyl group, a 2'-substituent group, or a protected 2'-substituent group;T.sub.1 is an acid labile hydroxyl protecting group; andT.sub.2 is a base labile hydroxyl protecting group or a covalent attachment to a solid support;(b) deblocking said acid labile hydroxyl protecting group to form a deblocked hydroxyl group;(c) treating said deblocked hydroxyl group with a further compound having the formula: [ STR 10 ] rein:Bx is a heterocyclic base moiety;R.sub.1 is H, a protected hydroxyl group, a 2'-substituent group, or a protected 2'-substituent group;T.sub.3 is a protected hydroxyl group, a nucleoside, a nucleotide, an oligonucleoside or an oligonucleotide; andT.sub.4 is a reactive phosphite group;and an activating agent for a time and under conditions effective to form an extended compound;(d) treating said extended compound with a capping agent to form a capped compound;(e) optionally treating said capped compound with an oxidizing agent to form an oxidized intermediate;(f) optionally repeating steps (b), (c), (d), and (e) to form a protected oligomeric compound, provided that at least one of said T.sub.4 has the formula: [ STR 11 ] rein:each R.sub.2, R.sub.3 and R.sub.4 is, independently, H, C.sub.1 -C.sub.6 alkyl, C.sub.1 -C.sub.6 alkenyl or C.sub.1 -C.sub.6 alkynyl;each R.sub.5 and R.sub.6 is, independently, H, C.sub.1 -C.sub.6 alkyl, C.sub.1 -C.sub.6 alkenyl, C.sub.1 -C.sub.6 alkynyl or C.sub.6 -C.sub.14 aryl; andR.sub.7 is a leaving group; and(g) treating said protected oligomeric compound with a solution of concentrated ammonium hydroxide containing a compound of the formula: EQU J.sub.1 --S--J.sub.2 rein:each J.sub.1 and J.sub.2 is, independently, H, alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, or substituted aralkyl; wherein such substitution can be amino, halogen, hydroxyl, keto, carboxyl, nitro, nitroso, nitrile, trifluoromethyl, trifluoromethoxy, O-alkyl, S-alkyl, NH-alkyl, N-dialkyl, O-aryl, S-aryl, NH-aryl, O-aralkyl, S-aralkyl, NH-aralkyl, amino, N-phthalimido, imidazolyl, azido, hydrazino, hydroxylamino, isocyanato, aryl, heterocyclyl, or carbocyclyl;with the proviso that one of J.sub.1 and J.sub.2 must be hydrogen;to form said oligomeric compound. [15.] A method for removing an allyl protecting group from an oligonucleotide comprising contacting said allyl protecting group with a mixture of concentrated ammonium hydroxide and a mercapto compound. The method of claim 15 wherein said mercapto compound is of formula: EQU J.sub.1 --S--J.sub.2 rein:each J.sub.1 and J.sub.2 is, independently, H, alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, or substituted aralkyl; wherein the substitutions are amino, halogen, hydroxyl, keto, carboxyl, nitro, nitroso, nitrile, trifluoromethyl, trifluoromethoxy, O-alkyl, S-alkyl, NH-alkyl, N-dialkyl, O-aryl, S-aryl, NH-aryl, O-aralkyl, S-aralkyl, NH-aralkyl, amino, N-phthalimido, imidazolyl, azido, hydrazino, hydroxylamino, isocyanato, aryl, heterocyclyl, or carbocyclyl with the proviso that one of J.sub.1 and J.sub.2 must be hydrogen. The method of claim 15 wherein said compound of formula J.sub.1 --S--J.sub.2 is mercaptoethanol, thiocresol, benzyl mercaptan, thiophenol, cysteine, cysteamine, glutathione or dithiothreitol. The method of claim 15 wherein the concentration of said compound of formula J.sub.1 --S--J.sub.2 in said solution of concentrated ammonium hydroxide is from about 1% to 50%. The method of claim 15 wherein the concentration of said compound of formula J.sub.1 --S--J.sub.2 in said solution of concentrated ammonium hydroxide is from about 1% to 30%.