C - Chemistry – Metallurgy – 07 – H
Patent
C - Chemistry, Metallurgy
07
H
260/210, 260/229
C07H 23/00 (2006.01) C07H 19/04 (2006.01) C07H 21/00 (2006.01)
Patent
CA 1194860
ABSTRACT A method for preparing a substrate capable of under- going catalytic-induced hydrolysis of the phosphate ester at the 3' position, to yield a species capable of being monitored spectrophotometrically or fluorometrically and thus useful in carrying out various non-isotopic immunoassays, comprises (a) forming a mononucleotide 2', 3' cyclic phosphate of the formula: Image wherein 8 is a nucleotide base, and wherein the CH20H group at the 4'-position is either cis or trans to the cyclic phosphate; (b) blocking said mononucleotide 2', 3'-cyclic phos- phate at the 5'-hydroxyl with a suitable 5'-blocking member to form a mononucleotide 5'-O-blocked-2', 3'-cyclic phosphate; (c) opening said cyclic phosphate so that essentially only a mononucleotide 5'-O-blocked-2'-hydroxyl-3'-phosphate is formed under conditions leaving the 5'-position essentially blocked (d) blocking said mononucleotide 5'-O-blocked 3'- phosphate at the 2'-hydroxyl with a suitable 21-O-blocking mem- ber to form a mononucleotide 2'-blocked-5'-blocked-3'-phosphate; and (e) forming a 2'-O-blocked-5'-O-blocked phosphodi- ester by bonding said mononucleotide 2'-O-blocked-5'-O-blocked- 3'-phosphate with a moiety selected from the group consisting of a chromophore or fluorophore.
397663
Kang Jemo
Tolman Glen L.
Osler Hoskin & Harcourt Llp
Roche Diagnostics Corporation
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