SU1041547A1 - Process for removing cyanethyl protective group from derivatives of mono- and dinucleotides - Google Patents

Abstract

METHOD OF REMOVING A CYANETHYL PROTECTIVE GROUP WITH DERIVATIVE MONOUS DYNUCLEOTIDES OF THE GENERAL FORM OO IIII Rd (K O-RO-O) O-RO-O-CH CHjCN, I OR OR 0 protected group; chlorophenyl group; or 1 by treating them with an organic base in an organic solvent medium, characterized in that, in order to intensify the process, a 2% solution of pentamethylguanidine in anhydrous diode is used as an organic base.

Description

SP

"Vi The invention relates to the chemical synthesis of nucleic acid fragments, namely to an improved method for removing the terminal cyan ethyl protective phosphate group from derivatives of mono and dinucleotides of the general formula 0 O (-O) N -0- / i-0-CH 1 I 22 OR OR (i; where M is a nucleoside that is protected by a heterocyclic nucleus; R-Mons5 methoxytrityl or dimethoxytritol; 5-0 protective group; R is a chlorophenyl group or 1. A method is known for removing the cyanoethyl protective group based on the treatment of mono- .and oligonucleotides inorganic solutions base in anhydrous and aqueous organic solvents. However, this method can lead to partial removal of other protective groups and hydrolysis of labile bonds, and therefore requires chromatographic purification of the target product. The closest to the proposed technical essence is achieved The method of removing the cyanoethyl group involves the treatment of mono- and oligonucleotides of formula (1) with an organic base in organic solvent, 50% solution of triethylamine in a cetonitrile. The output is quantitative. The duration of the process is 1 hour at room temperature. The disadvantage of this method is (with the duration of the process. The purpose of the invention is to intensify the process. The goal is achieved by the method of removing the cyanoethyl protecting group from the mono and dinucleotide derivatives of the general formula (I) by processing them with an organic base in the middle of an organic solvent, as an organic base, use a 2% solution of pentamethylguanidine in anhydrous dioxane. The yield is quantitative. process 3 min. Example 1. Preparation of 3-O-di-methoxytrityl-N-benz oyl Deoxo-Sicitidin-3-O- (p-chlorophenyl L-phosphate. To 40 µmol 5-0-dimethoxytrityl-N-benzoyldesoxycytidine-3 -0- (p- 20 μl of pure pentamethylguanidine is added to 1 ml of anhydrous dioxane in 1 ml of anhydrous dioxane. The mixture is kept at room temperature for 3 minutes and evaporated in vacuo. The yield of the chromatographically pure product is quantitative. The product is analyzed by liquid chromatography on silanized silica gel in a system 30 -60% aqueous acetonitrile in 0.05 M ac Tate triethylammonium. PRI m e-p 2. Decyanethylation of the polymer-5-0-dimethoxytrityldesoxythymidine-z -0- (p-chlorophenyl) cyanoethyl phosphate. To 200 mg of a polystyrene polymer containing 40 µmol of covalently bound 5-0-dimerxytrityl-desoxythymidine-3-O- (m-chlorophenyl) cyanoethyl phosphate, add 1 ml of anhydrous dioxane, degas the solution and add 20 µl of pure guanidine pentamethyl-dioxane . After 3 minutes, the polymer is filtered off. For checks on the completeness of decyanation of the polymer-nucleotide process. 1% solution triperoxonane acid in chloroform (2-20 ml, 2 min), washed with dioxane (310 ml). The combined extracts are evaporated, dissolved in 10 ml of 1p% dioxane, and chromatographed on silanized silica gel in the system with 15-50% aqueous acetonitrile. In 0.05 M triethylammonium acetate. The yield of deoxythymidine-3-0- (p-chlorophenyl) phosphate is quantitative .. Example 3. Preparation of 5-0-mo-. nomethoxytrityldesoxythymidilyl (3–5) desoximemitidine-3, 3-n (chlorophenyl phosphate. 20 μl of pure guanidine pentil etyl. The reaction mixture is aged at room temperature. 3 minutes, diluted with 20 ml of 0.05 .. M of triethylammonium and analyzed by liquid chromatography in the system 30-60% aqueous acetonitrile in 0.05 M triethylammonium acetate. The yield of the product decyanethyl is quantitative. Example 4. Decyanethylation of 5-0-dimethoxytrityl deoxycytidilyl (3–5) deosoxide anosine-3-0- (n-chlorophenyl) cyanethyl phosphate. A mixture of 30 µmol 5-0-dimethoxytrityl deoxycytidylylS3 5) deoxy-i adenosine-3-0- (p-chlorophenyl) cyanethylfrsfate, 0.75 ml of anhydrous dioxia and 15 µl of pure pentamethylguanidine is kept for 3 min, diluted with 20 ml of 0.05 M acetate triethylammonium and chromatographed on silanized blue glycol in the system 30-60% aqueous acetonitrile in 0.05 M zyethylammonium acetate. The yield of 5-0-dimethoxytrityl deoxycytidilyl (3–5) deoxy oxido 6-sin-3-0- (h-chlorophenyl) phosphate is 98.7%.

3. 1041547.

Etc. and measure 5. Preparation of 5 -0-lipata is dissolved in 0.75 ml of anhydrous methoxytrityl dophoxythymidyl (3-5) dioxane, 15 MKJT of pure deoxyguanosin-3-0- (i-chlorophenyl) pentamethylguanidine are added, the phosphate is kept . .3 min and 5-0-dimethoxytrityldehyde acetonitrile in 0.05 M aceoxythymidyl (3 - 5) triethylammonium deoxyguatate is analyzed by the method of liquid mol 30 μmol. The yield of the product nosine-3-0- (and-chlorofenyl) cyanoethylphosphate is quantitative.

Noah chromatography in the system 30-60%

Claims (1)

METHOD FOR REMOVING A CYANETHY PROTECTIVE GROUP FROM MONO- DERIVATIVES And dinucleotides of the general formula Rd (N * 0-P-0) _n li- OPO-CH ₂ CH ₂ CN, OROR where N * is a nucleoside protected at the heterocyclic nucleus, R is methsomethoxytrityl or dimethoxytrityl 5 ^{g is a} 0-protecting group; R * is a chlorophenyl group; n = 0 or 1 by treating them with an organic base in an organic solvent medium, characterized in that, in order to intensify the process, a 2% solution of pentamethylguanidine in anhydrous dioxane is used as an organic base. 4 ^ Si 4 * M