Phosphoramidite compounds and their use in producing oligonucleotides

Claim 2

A compound represented by one of the formulae: wherein B is a nucleoside or deoxynucleoside base; A is H, OH or OP. in which R 4 is a blocking group; R is a blocking group; R 1 is a hydrocarbyl radical containing up to about 10 carbon atoms; and X is NR 2 R 3 ', wherein R 2 ' and R' 3 taken sepsrately each represent alkyl, aryl, aralkyl, cycloalkyl and cycloalkylalkyl containing up to 10 carbon atoms; and R' 3 when taken together form an alkylene chain con- taining up to 5 carbon atoms in the principal chain and a total of up to 10 carbon atons with both terminal valence bonds of said chain being attached to the nitrogen atom to which R' 2 and R' 3 are attached: and R' 2 and when taken together with the nirrogen atom to which they are attached form a saturated nitrogen hecerocycle including at least one additional hetcroatom from the group consisting of nitrogen, oxygen and sulfur.

Claim 3

A compound according to Claim 1 wherein R is a trityl group or a di-p-anisylphenylmethyl group or a p-anisyldjLphenylmethyl group or p-anisyl-1-naphthylphcnyl- methyl or di-0-anisyl-1-naphthylmethyl or p-tolyldiphenyl- methyl.

Claim 4

A ccmpound according to Claim 1 or 2 wherein X is a saturated nitrogen heterocyclic, such as piperdine, morpnoline, and piperizine.

Claim 5

A compound according to any of Claims 1-3 wherein B is adenine, guanine, cytosine, uracil and thymine.

Claim 6

A compound according to Claim 1 wherein R is di-p-anisylphenylmethyl, B is 9-(N-6-benzoyladeninyl), R' 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is thyminyl, R' 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphcnyl-: methyl, B is 1-cytosinyl, R 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is 9-(N-6-benzoyladeninyl), R' 1 is methyl, A is H and X is piperidino, or R is di-p-anisylphenylmethyl, B is 9- guaninyl, R' 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is 9-(N-6-benzoyladeninyl), R' 1 is methyl, A is H and X is morpholino, or R is di-p-anisylphenylmethyl, B is thyminyl, R' 1 is methyl, A is H and X is morpholino, or R is di-p-anisylphenylmethyl, B is 1-cytosinyl, R 1 is methyl, A is H and X is morpholino, or R is di-p-anisylphenylmethyl, B is 9-guaninyl, R' 1 is methyl A is H and X is morpholino.

Claim 7

A compound according to Claim 5 wherein B is 1-(N-4-acetylcytosine), l-(N-4-benzoylcytosine), or l-(N-4-isobutyrylcytosine, or B is 9-(N-6-benzoyl- guaninyl), 9-(N-6-acetylguaninyl), or 9-(N-6-isobutyryl- guaninyl), or B is 9-(N-6-acetyladeninyl) or 9-(N-6-isobutyryladeninyl).

Claim 8

A compound according to Claim 1 wherein the compound is 5'-0-di-p-anisylphenylmethyl-N-isobutyryl- deoxyquanosine-3'-N,N-dimethylaminomethoxyphosphine:-or 5'-0-p-anisyl-1-naphthylphenylmcthyldeoxythymidinc-3'-N,N-dimethylaminomethoxyphospine; or 5'-o-di-c-anisyl-1- naothylraethyl-N-benzoyldeoxycytidine-3'-N,N-dimethylaminomethoxyphosphine; or 5'-o-p-tolydiphenylmethyl-N-benzoyl- deoxyadcnosinc-3'-N,N-diethlaminomcthoxyphosphine: or 5'-o-di-p-anisylphenylmethyl-N-isobutyryldeoxyquanosine-3'-N,N-morpholinomethoxyphosphine; or 5'-0-p-anisyl-1-naphthy]phenylmcthyldeoxythymidine-3'-N,N-morpholino- methoxyphosphine; or 5'-0-di-o-anisyl-1-naothylmethyl-N-benzoyldeoxycytidine-3'-N,N-:norpholinomethoxyphosphine; or 5'-0-p-tolydiphenylmethyl-N-benzoyldeoxyadenosine-3'-N,N-morpholinomethoxyphosphine; or 5'-0-di-p-anisylphenylmethyl-N-acetyldeoxyquanosine-3'-N,N-dimethylaminomethoxy- phosphinc; or 5'-O-di-p-anisylphenylmethyl-N-benzoyldeoxy- quanosine-3'-N,N-dimethylaminomethoxyphosphine; or 5'-O-di-o-anisyl-1-naphthylinethyl-N-acetyldeoxycytidine-3'-N,N-dimethyl- aminomcthoxyphosphine:or 5'-O-di-o-anisyl-1-naphthylmethyl-N-isobutyryldeoxycytidine-3'-N,N-dimethylaminomethoxyphosphine; or 5'-0-p-tolydiphenylmethyl-N-acetyldeoxyadenosine-3'-N,N-dimethylaminomethoxyphosphine; or 5'-0-p-tolydiphenylmethyl-N-isobutyryladenosine-3'-N,N-dimethylaminomethoxyphospine; or 5'-0-di-p-anisylphenylmethyl-N-acetyldeoxyquanosine-3'-N,N-morpholino- methoxyphosphine: or 5'-O-di-p-anisylphenylmethyl-N-benzoyldeoxyquanosine-3'-N,N-morpholinomethoxyphosphine; or 5'-0-di-o-anisyl-1-naothylmethyl-N-isobutyryldeoxy- cytidine-3'-N,N-morpholinomethoxyphosphine: or 5'-O-di-o-ansiyl-1-napthylmethyl-N-acetylddoxycytidine-3'-N,N-morpholinomethoxyphosohine;or 5'-O-p-tolydiphenylmethyl-N-acetyldeoxyadcnosine-3'-N,N-morpholinomethoxyphosphine; or 5'-O-p-tolydiphenylmethyl-N-isobutyryldeoxy- adenosine-3' -N,N-mornholinomethoxypnosphins.

Claim 9

A process of producing oligonucleotides which comprises the step of condensing the 3'-OH or 5'-OH group of a nucleoside or oligonucleotide with a nucleoside phosphite chracterized in that the nucleoside phosphite is a compound of one of the formulae: wherein B is a nucleoside or deoxynucleotide base; a is H, OH or OR' 1 in which R. is a blocking group; R is a blocking grcup; R' 1 is a hydrocarbyl radical containing up to about 10 carbon atoms; and X is NR' 2 R' 3 , wherein R' 2 and R' 3 taken separately each represent alkyl, aryl, aralkyl, cycloalkyl and cycloalkylalkyl containing up to 10 carbon atoms; R' 2 and R 3 when taken together form an alkylene chain con- taining up to 5 carbon atoms in the principal chain and a total of up to 10 carbon atoms with both terminal valence bonds of said chain being attached to the nitrogen atom to which R 2 and R 3 are attached; and R' 2 and R' 3 when taken together with the nitrogen atoni to which they are attached form a saturated nittogen hertercoycle including at least one additional heteroatom from the group consisting of nitrogen, oxygen and sulfur.

Claim 10

A process of producing oligonucleotides which comprises the step of condensing the 3'-OH or 5'-OH group of a nucleoside or oligonucleotide with a nucleoside phosphite chracterized in that the nucleoside phosphite is a compound of one of the formulae: wherein B is a nucleoside or deoxynucleotide base; a is H, OH or OR' 1 in which R. is a blocking group; R is a blocking grcup; R' 1 is a hydrocarbyl radical containing up to about 10 carbon atoms; and X is NR' 2 R' 3 , wherein R' 2 and R' 3 taken separately each represent alkyl, aryl, aralkyl, cycloalkyl and cycloalkylalkyl containing up to 10 carbon atoms; R' 2 and R 3 when taken together form an alkylene chain con- taining up to 5 carbon atoms in the principal chain and a total of up to 10 carbon atoms with both terminal valence bonds of said chain being attached to the nitrogen atom to which R 2 and R 3 are attached; and R' 2 and R' 3 when taken together with the nitrogen atoni to which they are attached form a saturated nittogen hertercoycle including at least one additional heteroatom from the group consisting of nitrogen, oxygen and sulfur.

Claim 11

The process according to Claim 8 wherein R is a trityl group, or a di-p-anisylphenylmethyl, or p-anisyldiphenylmethyl.

Claim 12

The process according to Claims 8 or 9 wherein X is a saturated nitrogen heterocyclic, such as piperidine, morpholine and piperizine.

Claim 13

The process according to any of Claims 8-10cohercin R is di-p-anisylphenylmethyl, B is 9-(N-6-benzoyladeninyl), R' 1 is methyl, A is H and X is morpholino, or R is di-p-anisylphenylmdthyl, B is thyminyl, R' 1 is methyl A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is 1-(N-4-benzoylcytosinyl), R' 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is 9-(N-6-benzoyladeninyl), R' 1 is methyl, A is H and X is dimethylamino, or R is di-p-anisylphenylmethyl, B is 9-(N-2-isobutyrylquaninyl), R' 1 is methyl, A is H and X is dimethylamino.

Claim 14

12 . The process according to any of Claims 8-11 wherein said nucleoside or oligonucleotide is covalently bonded to an inorganic polymer, for example, linked to the inorganic polymer through a base hydrolyzable covalent bond, e.g., an ester linkage.