SYNTHESIS METHOD OF 2'-O-(2-METHOXYETHYL) THIOPHOSPHATE OLIGONUCLEOTIDE Russian patent published in 2020 - IPC C12N15/113 C12N15/10 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, particularly to a novel method of producing antisense compounds which can be used as pharmaceutical substances for ameliorating the symptoms of spinal muscular atrophy. Disclosed method of producing modified thiophosphate oligonucleotide containing 2'-O-(2-methoxyethyl) ribonucleotides of structure UCACUUUCAUAAUGCUGG, where each nucleoside contains 2 ' -O- (2-methoxyethyl) modified sugar residue provides for synthesis of oligonucleotide on a solid-phase support using an amidophosphite method. To obtain thiophosphate groups, 3-[(dimethylaminomethylene)amino]-3H-1,2,4-dithiazol-5-thione (DDTT) is used in 30 % solution of pyridine in acetonitrile, release is performed by adding a saturated ammonia solution in the presence of disodium salt of ethylenediaminetetraacetic acid for 3 days at room temperature. Method then includes successively purifying oligonucleotides by ion exchange chromatography on Tosoh Super-Q 5PW column using Tris-HCl buffer, pH 7.0, with addition of 10 % acetonitrile in water, and then Tris-HCl buffer, pH 7.0, with addition of sodium perchlorate and 10 % acetonitrile in water, by reversed-phase chromatography on Waters xBridge WEN C18 column using a buffer, which is a solution of triethylammonium acetate in water, pH 7.0, and then a buffer which is a solution of triethylammonium acetate in 80 % aqueous solution of acetonitrile, pH 7.0, and gel filtration followed by extraction of the end product with drying by lyophilisation.

EFFECT: invention enables to obtain a product with higher degree of purification.

1 cl, 8 dwg, 4 tbl, 2 ex