Oligonucleotide labeling reactants based on acyclonucleosides and conjugates derived thereof

A labeling reactant of formula (I), useful for labeling an oligonucleotide using solid phase chemistry,

• 20
• wherein
• R is a temporary protecting group such as 4,4′dimethoxytrityl (DMTr), 4-methoxytrityl (MMTr), trityl (Tr), (9-phenyl)xanthen-9-yl (pixyl) or not present;
• A is either a phosphorylating moiety
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• where
• L is O, S, or not present
• L′ is H, XCH2CH2CN or XAr, where Ar is phenyl or its substituted derivative, where the substituent is nitro or chlorine, and X is O or S;
• L″ is O−, S−, Cl, N(i-Pr)2;
• or
• A is a solid support, preferably controlled pore glass or polystyrene, tethered to Z via a linker arm E″;
• Z is a bridge point and is formed from
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• E is a linker arm between R and Z, E′ is a linker arm between Z and Z′, E″ is a linker arm between Z and A and E′″ is a linker arm between Z′ and G, same or different, and is formed of one to ten moieties, each moiety being selected from the group consisting of phenylene, alkylene containing 1-12 carbon atoms, ethynediyl (—C≡C—), ether (—O—), thioether (—S—), amide (—CO—NH—, —CO—NH—, —CO—NR′— and —NR′—CO—), carbonyl (—CO—), ester (—COO— and —OOC—), disulfide (—S—S—), diaza (—N═N—), and tertiary amine (—N—R′), wherein R′ represents an alkyl containing less than 5 carbon atoms;
• Z′ is a purine or pyrimidine base selected from the group consisting of adenine, guanine, cytosine, thymine, uracil, 7-deazaadenine, 7-deazaguanine and hypoxantene, where E′ is attached to N1 of pyrimidines and N9 of purines, and E′″ is attached to C7 of 7-deazapurines, N3 or C5 of uracil, N3 of thymine, C5 or N4 of cytosine, C8, N2, N3 or O6 of guanine, C8, N6 or C2 of adenine, and where the exocyclic functional groups of said base are appropriately protected preferably with benzoyl, isobutyryl or acetyl, or
• Z′ is selected from the group consisting of imidazole, pyrazolo[3,4-d]pyrimidine, 4-amino-pyrazolo[3,4-d]pyrimidine, 1,2,4-triazine-3,5-dione, 5-amino-1,2,4-triazine-3-one, where E′ is attached to N1 of imidazole, N2 of 1,2,4-triazine-3,5-dione and 5-amino-1,2,4-triazine-3-one, and N7 of 4-amino-pyrazolo[3,4-d]pyrimidine and pyrazolo[3,4-d]pyrimidine, and E″ is attached to C4 or C5 of imidazole, C2 or C9 of 4-amino-pyrazolo[3,4-d]pyrimidine, C2, C4 or C9 of pyrazolo[3,4-d]pyrimidine, N4, C5 or C6 of 1,2,4-triazine-3,5-dione and N5 or C6 of N5-amino 1,2,4-triazine-3,5-dione and where the exocyclic functional groups of said base are appropriately protected preferably with benzoyl, isobutyryl or acetyl;
• G is a protected bivalent aromatic structure, tethered to two iminodiacetic acid ester groups N(CH2COOR″)2, where
• R″ is an alkyl of 1 to 4 carbon atoms, allyl, ethyltrimethylsilyl, phenyl or benzyl, which phenyl or benzyl can be substituted or unsubstituted, and one of the hydrogen atoms is substituted with E′″, and
• said bivalent aromatic structure is capable of absorbing light or energy and transferring the excitation energy to a lanthamide ion after said labeling reactant has been deprotected and converted to a lanthamide chelate, or
• G is a structure selected from the group consisting of
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• where
• R″ is an alkyl of 1 to 4 carbon atoms, allyl, ethyltrimethylsilyl, phenyl or benzyl, which phenyl or benzyl can be substituted or unsubstituted, and one of the hydrogen atoms is substituted with E′″, or
• G is a protected functional group, where the functional group is amino, aminooxy, carboxyl, thiol, and the protecting group is pthaloyl, trityl, 2-(4-nitrophenylsulfonyl)ethoxycarbonyl, fluorenylmethyloxycarbonyl, benzyloxycarbonyl, t-butoxycarbonyl or trifluoroacetuyl for amino and aminooxy, alkyl for carbonyl and alkyl or trityl for thiol, or
• G is a protected or unprotected organic dye, hapten or a spin label.