RU2014134383A - OLIGONUCLEOTIDES WITH MODIFIED PHOSPHATE GROUP AND METHOD FOR PRODUCING THEM - Google Patents

Abstract

1 A compound containing a modified phosphate group according to formula (I) (I) wherein the Z substituent is selected from —O, —S, —Se, —BH or —N (R), where R is H, C 1-6 alkyl or a protecting group; substituent X selected from the group comprising the 5'-end of a nucleoside, nucleoside analog, oligonucleotide, a modified oligonucleotide or oligonucleotide analog; Deputy Y is selected from the group comprising the 3'-end of a nucleoside, nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide or oligonucleotide analog; -H, -OH, -SH, -NHR, -O-PG or S-PG, where PG is a protecting group; a linker, a monophosphate group or a diphosphate group, or a label or a fluorescence quencher; or the Y substituent is selected from the group consisting of the 3 ′ end of a nucleoside, a nucleoside analog, an oligonucleotide, a modified oligonucleotide or an oligonucleotide analog; a substituent X is selected from the group comprising a 5 ′ end a nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide or oligonucleotide analog; -H, -OH, -SH, -NHR, -O-PG or S-PG, where PG is a protecting group; a linker, a monophosphate group or a diphosphate group, or a label or a fluorescence quencher; R, R and R substituents are independently selected from the group consisting of —H, —NRR, —OR, —SR, —S (O) R, —S (O) R , -S (O) NH, -S (O) NHR, -S (O) NR, -Calkyl, -Calkenyl, -Calkynyl, -Caryl or -Cheteroaryl; where each of the substituents R, R, R, R and R is independently selected from the group consisting of: -H, -Calkyl, -Calkenyl, -Calkynyl, -Caryl or -Cheteroaryl; where, in addition, R and R together form an alkylene or heteroalkylene chain 2-4 atoms long; where in addition R and R together with the atom to which they are bonded, form a 5-8 membered g terotsikl; further wherein r and together with the atoms to which they are attached, form a 5-8 membered heterocycle; wherein each alkyl, alkenyl, alkynyl, Ari

Claims (24)

1 Compound containing a modified phosphate group corresponding to formula (I)

(I) wherein substituent Z is selected from ^{-O -, -S -, -Se -} , -BH ₃ ^- or -N (R ^{N) -,} where R ^N is H, C _1-4 alkyl or a protective group; Deputy X is selected from the group consisting of the 5'-end of a nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide, or oligonucleotide analog; Deputy Y is selected from the group consisting of the 3'-end of a nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide or oligonucleotide analog; -H, -OH, -SH, -NHR ^N , -O-PG or S-PG, where PG is a protecting group; a linker, a monophosphate group or a diphosphate group, or a label or fluorescence quencher; or Deputy Y is selected from the group consisting of the 3'-end of a nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide or oligonucleotide analog; Deputy X is selected from the group consisting of the 5'-end of a nucleoside, nucleoside analog, oligonucleotide, modified oligonucleotide, or oligonucleotide analog; -H, -OH, -SH, -NHR ^N , -O-PG or S-PG, where PG is a protecting group; a linker, a monophosphate group or a diphosphate group, or a label or fluorescence quencher; the substituents R, R ¹ and R ^{2 are} independently selected from the group consisting of —H, —NR ^2A R ^2B , —OR ³ , —SR ³ , —S (O) R ³ , —S (O) ₂ R ³ , —S (O) ₂ NH ₂ , -S (O) ₂ NHR ³ , -S (O) ₂ NR ³ ₂ , -C _1-10 alkyl, -C _2-10 alkenyl, -C _2-10 alkynyl, -C _{6 -10} aryl or -C _5-10 heteroaryl; where each of the substituents R ^1A , R ^1B , R ^2A , R ^2B and R ³ independently selected from the group including: -H, -C _1-10 alkyl, -C _2-10 alkenyl, -C _2-10 alkynyl, -C _6-10 aryl or -C _5-10 heteroaryl; where in addition R ^1A and R ^2A together form an alkylene or heteroalkylene chain with a length of 2-4 atoms; where, in addition, R ^1A and R ^1B together with the atom to which they are attached form a 5-8 membered heterocycle; where additionally R ^2A and R ^2B together with the atom to which they are bonded form a 5-8 membered heterocycle; where each alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylene or heteroalkylene may be further substituted. 2. The compound of claim 1, wherein R, R ^1, and R ^{2 are} independently selected from the series —NR ^1A R ^1B , —OR ³ , —SR ^3, or —R ³ . 3. The compound of claim 2, wherein R is —NR ^1A R ^1B . 4. The compound of claim 1, wherein R ¹ is —R ³ , —NR ^2A R ^2B , —SR ^3, or —OR ³ . 5. The compound of claim 1, wherein each of R ^1A , R ^1B , R ^2A , R ^2B and R ^{3 is} independently selected from the group consisting of —H and —C _1-4 alkyl, optionally substituted with one or more substituents from the series —F , -Cl, -Br, -I, -OH, -C _1-4 alkoxy, -NH ₂ , -NH (C _1-4 alkyl) and -N (C _1-4 alkyl) ₂ . 6. The compound of claim 4, wherein R ¹ is NR ^2A R ^2B . 7. A compound according to claim. 6, wherein R ^1A, R ^1B, R ^2A and R ^2B is a methyl group. 8. The compound of claim 1, wherein R ^1A and R ^2A together form an alkylene or heteroalkylene chain of 2-4 atoms, and R ^1B and R ^{2B are} each independently selected from the group consisting of —H and —C _1-4 alkyl. 9. The compound of claim 8, wherein R ^1A and R ^2A together form a —CH ₂ —CH ₂ - or —CH ₂ —CH ₂ —CH ₂ - chain, and R ^1B and R ^{2B are} independently selected from —H or methyl groups. 10. The compound according to claim 1, in which R ^1A and R ^1B together with the atom to which they are attached form a 5-8 membered heterocycle, preferably pyrrolidine. 11. The compound of claim 10, wherein R ^2A and R ^2B, together with the atom to which they are attached, form a 5-8 membered heterocycle, preferably pyrrolidine. 12. A method for producing a compound according to claim 1, comprising reacting a phosphorous acid derivative with an azidophosphonium derivative in the presence of a solvent or a mixture of two or more solvents and additionally a silylating reagent and / or base. 13. The production method of claim 12, wherein the phosphorous acid derivative is phosphite or H-phosphonate. 14. The method according to p. 12, characterized in that the organic solvent for the reaction is selected from the list including acetonitrile, acetone, tetrahydrofuran (THF), 1,4-dioxane, 1,2-dimethoxyethane (DME), diethylene glycol dimethyl ether (diglyme ), DMF, N, N-dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), tetramethylurea, hexamethylphosphoric triamide (HMFTA), sulfolane, ethyl acetate, 1,3-dimethyl-2-imidazolidinone, ethyl ether. 15. The method according to p. 12, characterized in that said base is selected from the group consisting of triethylamine, N, N-diisopropylethylamine (DIEA), N-methylmorpholine, N-ethylmorpholine, N-methylpyrrolidine, N-methylpiperidine, tributylamine, 2, 6-lutidine, 2,4,6-collidine, 1,4-diazabicyclo [2.2.2] octane (DABCO), N-methylimidazole (NMI), 4-dimethylaminopyridine (DMAP), 1,8-diazabicyclo [5.4.0 ] undecene-7 (DBU), 1,5-diazabicyclo [4.3.0] nonen-5 (DBN), 1,8-bis-dimethylaminonaphthalene, 1,5,7-triazabicyclo [4.4.0] decen-5 (TBD ), 7-methyl-1,5,7-triazabicyclo [4.4.0] decen-5 (MTBD), 1,1,3,3-tetramethylguanidine (TMG), 2-tert-butyl-1,1,3, 3-tetramethylguanidine, f sfazenovoe base. 16. The method according to p. 12, in which the azidophosphonium derivative is a compound corresponding to the formula

where each of the substituents R, R ^1A , R ^1B , R ^2A and R ^2B independently may be —H or additionally substituted C _1-10 alkyl; and / or where R ^1A and R ^2A together form an alkylene or heteroalkylene chain 2-4 atoms long, and R ^1B and R ^{2B are} each independently selected from the group consisting of —H and —C _1-4 alkyl; where R ^1A and R ^1B together with the atom to which they are attached form a 5-8 membered heterocycle; or R ^2A and R ^2B , together with the atom to which they are attached, form a 5-8 membered heterocycle. 17. The method according to p. 12, characterized in that the reaction is additionally carried out in the presence of a silylating reagent, which is selected from the group consisting of the following silylating agents: N, O-bis (trimethylsilyl) acetamide (BSA), N, O-bis (trimethylsilyl ) trifluoroacetamide (BSTFA), trimethylchlorosilane, trimethyl bromosilane, trimethyl iodosilane, hexamethyldisilazane, trimethylsilyl trifluoromethanesulfonate (TMSOTf), triethylchlorosilane, dimethylisopropylchlorosilane dimethyl-trimethylsilane-trimethylsilane-dimethyl-trimethylsilane-dimethyl-trimethylsilane-dimethyl-trimethylsilane-trimethylsilane-dimethyl-trimethylsilane-dimethyl ldihlorsilan, diphenyldichlorosilane. 18. The oligonucleotide according to claim 1, in which the phosphate group that binds residues of adjacent nucleosides or nucleoside analogs is a modified phosphate group corresponding to the formula according to claim 1. 19. The oligonucleotide according to claim 18, wherein the phosphate group that binds residues of adjacent nucleosides or nucleoside analogs is an N- (phosphoranilidene) phosphoramide group. 20. The use of an in vitro oligonucleotide according to claim 18. 21. The use of an in vitro oligonucleotide according to claim 18 as a diagnostic tool. 22. The oligonucleotide according to claim 18 for use in medicine as a medicine. 23. The use of the oligonucleotide according to claim 18 as part of a pharmaceutical composition comprising the oligonucleotide according to claim 18 and a pharmaceutically acceptable component or components for treating a disease. 24. The use of the oligonucleotide according to claim 18, comprising administering to the patient in need of treatment the oligonucleotide according to claim 18 for the treatment of a disease.